CN104363773A - Herbal composition for treatment of gastrointestinal inflammatory diseases and method to prepare and use thereof - Google Patents
Herbal composition for treatment of gastrointestinal inflammatory diseases and method to prepare and use thereof Download PDFInfo
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- CN104363773A CN104363773A CN201380026260.XA CN201380026260A CN104363773A CN 104363773 A CN104363773 A CN 104363773A CN 201380026260 A CN201380026260 A CN 201380026260A CN 104363773 A CN104363773 A CN 104363773A
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Abstract
The present invention relates to a herbal composition to treat gastrointestinal inflammatory diseases in a subject suffering therefrom. The present invention further provides a method to use a pharmaceutical effective amount of the herbal composition to treat Gl inflammatory diseases. Particularly, this invention relates to using a pharmaceutically effective amount of the herbal composition extract having the ability to inhibit the expression of pro-inflammatory cytokines, attenuate disruption of epithelial barrier, and modulate gut function
Description
related application
This application claims the priority that the sequence number submitted on May 19th, 2012 is the U.S. Provisional Application of 61/649,287, its whole disclosure is incorporated to herein by reference.
Technical field
The present invention relates to the herbal-composition (herbal composition) for treating gastroenteritis disease in the object suffering from gastroenteritis disease (gastrointestinal inflammatory disease).Present invention also offers and use the described herbal-composition of medicine effective quantity to treat the method for gastroenteritis disease.Especially, the present invention relates to ageratum (the HUOXIANG ZHENGQI using medicine effective quantity, HXZQ) extract, described extract can suppress the expression of pro-inflammatory cytokine, weaken the destruction of epithelial barrier (epithelial barrier) and regulate gut function (gut function).
Background technology
Ulcerative colitis (ulcerative colitis, UC) and Crohn disease (Crohn ' s diesease, CD) be two kinds of chronic inflammatory diseases, be referred to as inflammatory bowel disease (inflammatory bowel disease, IBD).According to estimates, 1,100,000 Americans that have an appointment suffer from IBD, and diagnose out about 30,000 new cases (Hanauer SB, 2006) every year.The feature of IBD is the chronic inflammation causing diarrhoea and abdominal pain.There is the risk rising of colorectal cancer in the patient suffering from long-term colitis.The relapsing course of IBD needs extended patient to nurse, and needs actively to carry out medicine or surgical intervention with inducer remission simultaneously.The cause of disease of IBD is not yet known, but thinks that it responds imbalance by the change of microorganism mattress group, the hereditary predisposition that disease occurs and mucosa-immune to cause.The process LAN of lymphocytic activation and inflammatory cytokine causes chronic enteritis.The research of exploitation novel treatment mainly concentrates on the biological reagent of the generation of the crucial Th1 cell factor of specific inhibition.The existing biopharmaceuticals for inflammatory bowel disease (IBD) (its object is to the generation blocking crucial Th1 cell factor) provides some benefits for patient.But these reagent is not generally suitable for and usually lost efficacy after long-term use.In addition, although these anti-Th1 therapeutic agents are that IBD patient provides some benefits, it can not generally be suitable for far away.First, they are not all effective to all IBD patients.Secondly, its generation due to the neutralizing antibody for this medicine and there is the trend lost efficacy in time.Finally, classical Th1 cell factor is important in defence intra-cellular pathogens, and the cell factor thus blocking these keys may make patient be in high risk that some infection occurs or reactivates.
For centuries, Chinese herbal treatment agent is used to treat stomach and intestine (gastrointestinal, GI) symptom, but its effect and most of potential mechanism it be unclear that.Traditional herb formulation comprises many different components usually, and these components act on multiple position/approach with the chemical reaction between potential synergy and component herbal medicine, thus can make effort maximization.
Ageratum is the Chinese medicine preparation deriving from ancient Chinese pharmacy books " formulary of peaceful benevolent dispensary " (Formularies of the Bureau of People ' s Welfare Pharmacies), and is used to treatment heatstroke and relevant gastrointestinal disease traditionally.Therefore, expect that research ageratum composition is to the effect of gastroenteritis disease, especially innate immune cells is produced to the effect of cell factor, ageratum composition blocks/weakens the ability of the barrier function of imbalance; And ageratum composition is to the effect regulating intestinal smooth muscle contraction and epithelial secretion.
summary of the invention
The object of this invention is to provide herbal-composition for treating GI inflammatory disease in the object suffering from GI inflammatory disease.Present invention also offers and use the described herbal-composition of medicine effective quantity to treat the method for GI inflammatory disease.
In one aspect, the invention provides the herbal-composition for treating GI inflammatory disease in object, described composition comprises the extract available from Pogostemon cablin (Pogostemonis Herba), perilla leaf (Perillae Folium), dried orange peel (Citri Reticulatae Pericarpium), the bighead atractylodes rhizome (Atractylodis Macrocephalae Rhizoma) and the root of Dahurain angelica (Angelicae Dahuricae Radix).
In the one of this embodiment is implemented, often kind of herbal ingredients unit range is by weight as follows: Pogostemon cablin accounts for about 18 % by weight to about 48 % by weight of composition; Perilla leaf accounts for about 6 % by weight to about 16 % by weight of composition; Dried orange peel accounts for about 12 % by weight to about 32 % by weight of composition; The bighead atractylodes rhizome accounts for about 12 % by weight to about 32 % by weight of composition, and the root of Dahurain angelica accounts for about 6 % by weight to about 16 % by weight of composition.
In the another kind of this embodiment is implemented, often kind of herbal ingredients unit range is by weight as follows: Pogostemon cablin accounts for about 28 % by weight to about 38 % by weight of composition; Perilla leaf accounts for about 9 % by weight to about 13 % by weight of composition; Dried orange peel accounts for about 17 % by weight to about 27 % by weight of composition; The bighead atractylodes rhizome accounts for about 17 % by weight to about 27 % by weight of composition, and the root of Dahurain angelica accounts for about 9 % by weight to about 13 % by weight of composition.
In the another kind of this embodiment is implemented, often kind of herbal ingredients units is by weight as follows: Pogostemon cablin accounts for about 33 % by weight of composition; Perilla leaf accounts for about 11 % by weight of composition; Dried orange peel accounts for about 22 % by weight of composition; The bighead atractylodes rhizome accounts for about 22 % by weight of composition, and the root of Dahurain angelica accounts for about 11 % by weight of composition.
In the another kind of this embodiment is implemented, herbal-composition is the extraction of essential oil thing from herbal ingredients of medicine effective quantity, and described essential oil is by carrying out steam distillation to extract forming the aqueous solution to herbal ingredients.
Present invention also offers herbal-composition, it includes the above-mentioned composition of effective amount and pharmaceutically suitable carrier, diluent or excipient or its combination.
In one embodiment, above-mentioned composition is pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
The present invention relates to and use the herbal-composition of medication amount to the Expression of Macrophages pro-inflammatory cytokine suppressing LPS to stimulate, weaken the epithelial barrier destruction that IFN γ induces, and regulate gut function.These activity contribute to the beneficial effect of the preparation resisting GI inflammatory disease or illness, described gastrointestinal disease or illness comprise inflammatory bowel disease (ulcer disease and Crohn disease), IBS, chylous diarrhea and opiate withdrawal syndrome (opiate withdrawal symptom), such as, suffer from diarrhoea.
In a preferred embodiment of the invention, provide a kind of method by treating GI inflammatory disease to the herbal-composition of subject effective amounts, described herbal-composition comprises the extract from Pogostemon cablin and perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica.
In in of this embodiment, described herbal-composition regulates the expression of pro-inflammatory cytokine in the object suffering from GI inflammatory disease, weakens the destruction to epithelial barrier, or regulates gut function.
In another embodiment, the herbal-composition of the described GI of being used for the treatment of inflammatory disease comprises available from Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome, the root of Dahurain angelica, the bark of official magnolia (Magnoliae Officinalis Cortex), the tuber of pinellia (Pinelliae Rhizoma), Poria cocos (Poria), Radix Glycyrrhizae (Glycyrrhizae Radix et Rhizoma), the shell of areca nut (Arecae Pericarpium), balloonflower root (Platycodonis Radix), the extract of ginger (Zingiberis Rhizoma Recens) and date (Jujubae Fructus).
In the one of this embodiment is implemented, often kind of herbal ingredients unit range is by weight as follows: Pogostemon cablin accounts for about 10 % by weight to about 20 % by weight of composition; Perilla leaf accounts for about 2.5 % by weight to about 7.5 % by weight of composition; Dried orange peel accounts for about 5 % by weight to about 15 % by weight of composition; The bighead atractylodes rhizome accounts for about 5 % by weight to about 15 % by weight of composition; The root of Dahurain angelica accounts for about 2.5 % by weight to about 7.5 % by weight of composition; The bark of official magnolia accounts for about 5 % by weight to about 15 % by weight of composition; The tuber of pinellia accounts for about 5 % by weight to about 15 % by weight of composition; Poria cocos accounts for about 2.5 % by weight to about 7.5 % by weight of composition; Radix Glycyrrhizae accounts for about 5 % by weight to about 15 % by weight of composition; The shell of areca nut accounts for about 2.5 % by weight to about 7.5 % by weight of composition; Balloonflower root accounts for about 5 % by weight to about 15 % by weight of composition; Ginger accounts for about 0.5 % by weight to about 2.5 % by weight of composition; And date accounts for about 1.5 % by weight to about 3.5 % by weight of composition.
In the another kind of this embodiment is implemented, often kind of herbal ingredients unit range is by weight as follows: Pogostemon cablin accounts for about 12.5 % by weight to about 17.5 % by weight of composition; Perilla leaf accounts for about 4 % by weight to about 6 % by weight of composition; Dried orange peel accounts for about 7.5 % by weight to about 12.5 % by weight of composition; The bighead atractylodes rhizome accounts for about 7.5 % by weight to about 12.5 % by weight of composition; The root of Dahurain angelica accounts for about 4 % by weight to about 6 % by weight of composition; The bark of official magnolia accounts for about 7.5 % by weight to about 12.5 % by weight of composition; The tuber of pinellia accounts for about 7.5 % by weight to about 12.5 % by weight of composition; Poria cocos accounts for about 4 % by weight to about 6 % by weight of composition; Radix Glycyrrhizae accounts for about 7.5 % by weight to about 12.5 % by weight of composition; The shell of areca nut accounts for about 4 % by weight to about 6 % by weight of composition; Balloonflower root accounts for about 7.5 % by weight to about 12.5 % by weight of composition; Ginger accounts for about 1 % by weight to about 2 % by weight of composition; And date accounts for about 2 % by weight to about 3 % by weight of composition.
In the another kind of this embodiment is implemented, the percentage by weight of often kind of herbal ingredients is as follows: Pogostemon cablin accounts for about 15 % by weight of composition; Perilla leaf accounts for about 5 % by weight of composition; Dried orange peel accounts for about 10 % by weight of composition; The bighead atractylodes rhizome accounts for about 10 % by weight of composition; The root of Dahurain angelica accounts for about 5 % by weight of composition; The bark of official magnolia accounts for about 10 % by weight of composition; The tuber of pinellia accounts for about 10 % by weight of composition; Poria cocos accounts for about 5 % by weight of composition; Radix Glycyrrhizae accounts for about 10 % by weight of composition; The shell of areca nut accounts for about 5 % by weight of composition; Balloonflower root accounts for about 10 % by weight of composition; Ginger accounts for about 1.5 % by weight of composition; And date accounts for about 2.5 % by weight of composition.
In a further preferred embodiment, the present invention relates to the herbal-composition for treating GI inflammatory disease in the object suffering from GI inflammatory disease, this herbal-composition is made up of at least the first extract of effective dose and the mixture of the second extract.Described first extract comprises the purified alcoholic solution extract of the herbal medicine bark of official magnolia.Described second extract comprise combine with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger and the purified water solution extract of date by carry out distilling to Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and root of Dahurain angelica component remove essential oil after the medicinal material residue that decocts together with date with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger.
In in of this embodiment, described first to extract be, with 60% ethanol, the bark of official magnolia is extracted 3 times, and then filter acquisition filtrate, merging filtrate is also condensed into the pasty state extract of clarification.
This embodiment another in, extraction of essential oil is from distillate Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica being carried out to steam distillation acquisition.The purified aqueous solution is left after extract essential oil from distillate.Medicinal material residue is obtained after steam distillation is carried out to above-mentioned herbal ingredients.
This embodiment another in, filter the second extract, filtrate be concentrated into suitable volume, centrifugal, and be condensed into the pasty state extract of clarification further.
This embodiment another in, the mixture of the first extract and the second extract and essential oil merge.
This embodiment another in, present invention also offers herbal-composition, it includes above-mentioned composition and pharmaceutically suitable carrier, diluent or the excipient of effective amount, or its combination.
In one aspect of the method, above-mentioned composition is pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
In a further preferred embodiment, present invention also offers the method for treating GI inflammatory disease in object, it comprises the herbal-composition to described subject effective amounts, and described herbal-composition comprises the extract from Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome, the root of Dahurain angelica, the bark of official magnolia, the tuber of pinellia, Poria cocos, Radix Glycyrrhizae, the shell of areca nut, balloonflower root, ginger and date.
In in of this embodiment, described GI inflammatory disease is selected from: intestinal disease, comprises ulcer disease and Crohn disease, IBS, chylous diarrhea, opiate withdrawal syndrome and diarrhoea.
This embodiment another in, described herbal-composition regulates the expression of pro-inflammatory cytokine in the object suffering from GI inflammatory disease, weaken the destruction of epithelial barrier or regulate gut function.
For those of ordinary skills, the remainder by presents is become obvious by these features of the present invention, advantage and other embodiments.
accompanying drawing is sketched
In order to understand the object of character of the present invention and expectation more fully, can with reference to following detailed description by reference to the accompanying drawings.
Figure 1A-E illustrates with after ageratum (HXZQ) extract-treated, the mrna expression of the inflammatory cytokine detected by PCR in real time.
Fig. 2 A-F illustrates with after the process of ageratum (HXZQ) essential oil, the mrna expression of the inflammatory cytokine detected by PCR in real time.
Fig. 3 A-E illustrates that ageratum (HXZQ) extract and essential oil weaken the effect that IFN γ destroys epithelial barrier.
Fig. 4 A-D illustrates the effect that ageratum (HXZQ) extract shrinks intestinal smooth muscle.
Fig. 5 A-C illustrates that ageratum (HXZQ) extract reduces enteric epithelium secretion.
detailed Description Of The Invention
By reference to following detailed description of the present invention, more easily the present invention can be understood.Should be understood that and the invention is not restricted to described concrete device, method, conditioned disjunction parameter herein, and term used, only for being described the object of specific embodiments by example, and is not intended to limit the present invention for required protection herein.In addition, the noun not having numeral-classifier compound to modify used in description and appended claims represents one/kind or more/kind, and comprises at least this particular value when mentioning concrete numerical value, except context is obviously otherwise noted.In this article, scope can be expressed as " about " or " approximately " particular value and/or to " about " or " approximately " another particular value.When illustrating such scope, another embodiment comprises a particular value and/or to another particular value.Similarly, when by using preceding " about " that value is expressed as approximation, should be understood that this particular value forms another embodiment.
Give the details of one or more embodiment of theme disclosed in the present invention in this document.After have studied presents and information be provided, will be apparent to those of ordinary skill in the art for the amendment of embodiment described in this file and other embodiments.The information provided in presents and particularly the detail of described exemplary mainly provide for being convenient to the clear object understood, and should not be construed as restriction.
Although think that those of ordinary skill in the art will understand following term well, but still provide and to help as given a definition to explain theme disclosed in the application.
Unless otherwise stated, all technical terms used herein and scientific terminology have usual the understood implication of theme one skilled in the art disclosed by the invention.Although will be described exemplary process, device and material now, all can be used for implementing or test theme disclosed in this invention with described herein those similar or that be equal to many methods, device and materials.
Following Patent Law convention for a long time, referring to " one or more " during the noun modified when using countless measure word in the application's (comprising claims).Therefore, when mentioning " polymer ", comprise multiple such polymer, by that analogy.
Unless otherwise stated, the numerical value of the amount of all expression compositions used in the present specification and claims, character (such as reaction condition) etc. is all interpreted as being modified by term " about " in all examples.Therefore, unless the contradiction of illustrating, otherwise the numerical parameter provided in the specification and claims be can according to expecting the character obtained by theme disclosed in this invention and the approximation changed.
As used in this article, when mentioning quality, weight, time, volume, when the value of concentration or percentage or amount, term " about " is intended to the following change comprising specified rate: be ± 20% in some embodiments, be ± 15% in some embodiments, be ± 10% in some embodiments, be ± 5% in some embodiments, be ± 2.5% in some embodiments, be ± 2% in some embodiments, be ± 1% in some embodiments, be ± 0.5% in some embodiments, and be ± 0.1% in some embodiments, these changes are suitable for carrying out disclosed method.Should also be understood that and there is disclosed herein much numerical value, and disclosed each numerical value is also disclosed as " about " this concrete numerical value except itself in this article.Such as, if disclose numerical value " 10 ", then also disclose " about 10 ".Should also be understood that each unit also disclosed between Liang Ge concrete unit.Such as, if disclose 10 and 15, then 11,12,13 and 14 are also disclosed.
Term used herein " treatment " includes but not limited to suppress the process of tissue damage, stop tissue produce damage generation, reduce the order of severity to tissue damage, improvement or the alleviation symptom relevant to tissue damage and reparation, regenerate and/or cause tissue damage or one or more resolution of symptoms relevant with tissue damage.
Term used herein " effective dose " refer to be enough to treat as this paper the disease amount that limits (such as, weaken the destruction to epithelial barrier in GI system) composition (such as, comprising the herbal-composition of the herbal ingredients of theme disclosed in the present invention).The actual amount of theme composition disclosed in the present invention can change to use the amount of composition of the Expected Response of effectively acquisition special object and/or be applied to specified disease.Selected amount will depend on many factors, comprise the activity of composition, preparation, route of administration, with the combination of other drug or therapeutic agent, the sanatory order of severity and accept the health of object for the treatment of and medication history before this.Determine and adjust treatment effective dose and evaluate when and how to make these adjustment be known for field of medicaments those of ordinary skill.
The present composition can comprise the drug moiety of the present invention of " treatment effective dose ".Term used " treatment effective dose " means in the scope of good medical judgment herein, compound or composition are (such as, HUXIANGHUANGQI extract or essential oil) be enough to the to be regulated or sanatory positive improvement of significantly induction but enough low to avoid the amount of side effect (if present, with rational benefit/risk than).The treatment effective dose of described compound or composition is by different according to following factor: the concrete illness for the treatment of, the age of terminal user and health, treat the factor such as/order of severity of the illness of prevention, duration for the treatment of, the character of concurrent treatment, the particular compound used or composition, concrete pharmaceutically suitable carrier of using.All percentage used herein is by weight, except being otherwise noted.
Pharmaceutical composition described herein itself can be applied to mammalian object, or uses with the form of the pharmaceutical composition of itself and other active component (as in therapeutic alliance) or suitable carrier or mixed with excipients." Remington ' s Pharmaceutical Sciences, " Mack Publishing Co. is found in, Easton, Pa., the 18th edition, 1990 for preparing and use the technology of preparation described in the application " in.Suitable route of administration such as can comprise per os, per rectum, thoroughly mucous membrane or intestines are used.
With regard to theme disclosed herein, preferably to liking vertebrate subject.Preferred vertebrate subject is warm-blooded vertebrate; Preferred warm-blooded vertebrate is mammal.Preferred mammal is most preferably people.Term used herein " object " comprises humans and animals two kinds of objects.Therefore, veterinary therapeutic use is provided according to theme disclosed in the present invention.Therefore, theme disclosed in the present invention for diagnosing mammal such as people, and because of those endangered and important mammals, such as northeastern tiger (Siberian tiger); There is the animal of Economic Importance, such as, the animal consumed for people of farm breeds; And/or people is had to the animal of social importance, such as raising pets or animal in zoo.The example of such animal includes but not limited to: carnivore, such as cat and dog; Pig (swine), comprises a pig (pig), pork pig (hog) and wild boar; Ruminant and/or ungulate, such as ox, bull (oxen), sheep, giraffe, deer, goat, wild ox and camel; And horse.Additionally provide the treatment to bird, comprise the bird of those kinds for the treatment of in endangered and/or raising at the zoo, and bird, more particularly, domestic bird (domesticated fowl), i.e. poultry (poultry), such as turkey, chicken, duck, goose, guinea fowl (guinea fowl) etc., because they also have Economic Importance to people.Therefore, additionally provide the treatment to domestic animal, described domestic animal includes but not limited to: pig, ruminant, ungulate, horse (comprise contest with horse), poultry etc.
Except as otherwise noted, otherwise the enforcement of theme disclosed in the present invention can adopt cell biology, cell chulture, molecular biology, transgcnic biology, microbiology, recombinant DNA and immunologic routine techniques, these technology are all in the scope that those skilled in the art know.These technology obtain in the literature to be explained in detail.See such as, Molecular Cloning A Laboratory Manual (1989), the 2nd edition, Sambrook edits; Fritsch and Maniatis edits, Cold Spring Harbor Laboratory Press, the 16th and 17 chapters; U.S. Patent No. 4,683,195; DNA Cloning, I and the II phase, Glover edits, 1985; Oligonucleotide Synthesis, M.J.Gait edits, and 1984; Nucleic Acid Hybridization, D.Hames & S.J.Higgins edits, and 1984; Transcription and Translation, B.D.Hames & S.J.Higgins edits, and 1984; Culture Of Animal Cells, R.I.Freshney, Alan R.Liss, Inc., 1987; Immobilized Cells And Enzymes, IRL Press, 1986; Perbal (1984), A Practical Guide To Molecular Cloning; Consult as the method in Publication about Document: Enzymology (Academic Press, Inc., N.Y.); Gene Transfer Vectors For Mammalian Cells, J.H.Miller and M.P.Calos edits, Cold Spring Harbor Laboratory, 1987; Methods In Enzymology, the 154th and 155 phases, the editors such as Wu, Academic Press Inc., N.Y.; (Mayer and Walker edits Immunochemical Methods In Cell And Molecular Biology, Academic Press, London, 1987; Handbook Of Experimental Immunology, the I-IV phase, D.M.Weir and C.C.Blackwell edits, and 1986.
Term " GI inflammatory disease " refers to that to be raised what cause by pro-inflammatory cytokine cyclical level take inflammation as enterogastric diseases or the illness of feature.Immunocyte (comprising T cell, macrophage and neutrophil cell) is the prime producer of these pro-inflammatory cytokines.GI inflammatory disease comprises intestinal disease (ulcer disease and Crohn disease), IBS, chylous diarrhea, opiate withdrawal syndrome and diarrhoea.
Term used herein " essential oil " or " volatile oil " refer to from having hydrophily this general herbal medicine of tendency and any concentrated volatile aromatic compounds of plant extract.
Term " decoction " herein refers to herbal medicine and water to be heated to high temperature, generally at 100 DEG C ± 5 DEG C, and the content of this heating is remained on the process of the temperature of 90 DEG C to 100 DEG C.
The present invention relates to herbal-composition ageratum (HXZQ) extract using medication amount, the Expression of Macrophages pro-inflammatory cytokine that this extract can suppress LPS-to stimulate, the destruction to epithelial barrier of weakening IFN γ induction and adjustment gut function.These activity contribute to the beneficial effect of this ageratum extract for treating GI inflammatory disease in pharmaceutical preparation or illness, and described GI inflammatory disease or illness comprise inflammatory bowel disease (ulcer disease and Crohn disease), intestinal irritable syndrome, chylous diarrhea and opiate withdrawal syndrome.
Herbal-composition ageratum (HXZQ) is the Chinese medicine preparation deriving from ancient Chinese pharmacy books " formulary of peaceful benevolent dispensary ", and is used to treatment GI illness traditionally.But the beneficial effect stated is anecdote (anecdotal) mainly, and confirms without science data.Therefore, the object of the invention is to study: (i) HXZQ extract and essential oil are on the impact being produced cell factor by the innate immune cells stimulated with inflammatory mediator; (ii) in individual layer T84 cell, HXZQ extract and essential oil block/weaken the ability of the barrier function of imbalance; (iii) HXZQ extract and essential oil are to the effect regulating intestinal smooth muscle contraction and epithelial secretion.
In an aspect, described preparation includes the ageratum extract of effective amount and essential oil and physiologically acceptable carrier, diluent or excipient, or its combination.Term " pharmaceutical composition " or " pharmaceutical preparation " refer to the mixture of ageratum extract and essential oil and other chemical constituents (such as diluent or carrier).Pharmaceutical composition is convenient to pharmaceutical preparation to be applied to object.
Compound ageratum extract and essential oil are incorporated in cell or tissue is convenient in term " carrier " definition.Such as, methyl-sulfoxide (DMSO) is conventional carrier.Term " diluent " is defined in dilution with water and will dissolves the compound of ageratum extract and/or essential oil.Use the salt be dissolved in buffer solution as diluent in the art.Term " physiology can accept " definition and not eliminate the biologically active of ageratum extract and essential oil and the carrier of characteristic or diluent.
Term used herein " pharmaceutically acceptable " means described carrier, diluent, excipient and/or salt must be compatible with other compositions of preparation, and harmless to its recipient.
Term used herein " pharmaceutically suitable carrier " means the solid of the nontoxic inertia of any type, semisolid, diluent, encapsulating material or formulation auxiliary agents.Some examples that can be used as the material of pharmaceutically suitable carrier are sugar, such as lactose, dextrose plus saccharose; Starch, such as cornstarch and farina; Cellulose and its derivates, such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; Fructus Hordei Germinatus; Gelatin; Talcum; And other nontoxic biocompatible lubricant, such as lauryl sodium sulfate and dolomol, and colouring agent, releasing agent, coating agent, sweetener, flavouring and aromatic, anticorrisive agent and antioxidant also can be present in described composition according to the judgement of makers-up.
For oral administration, can easily by ageratum extract and essential oil and pharmaceutically suitable carrier as known in the art be combined to prepare.Such carrier makes it possible to ageratum extract to be mixed with tablet, pill, sugar-coat agent, capsule, liquid agent, gel, syrup, slurries agent, supensoid agent etc., takes in for patient's per os to be treated.The ageratum extract used for per os and the pharmaceutical preparation of essential oil obtain by following steps: mix one or more of solid excipient; optionally grind gained mixture; processing granular mixture; add suitable adjuvant (if desired), to obtain tablet or dragee core.
Especially, suitable excipient is filler, and such as sugar, comprises lactose, sucrose, mannose or sorbose; Cellulose preparations, such as cornstarch, wheaten starch, rice starch, farina, gelatin, tragacanth, methylcellulose, HPMC, sodium carboxymethylcellulose and/or polyvinylpyrrolidone (PVP).If desired, disintegrant can be added, such as PVPP, agar, alginic acid or alginate such as sodium alginate.
Dosage and concentration can adjust according to actual conditions.Those skilled in the art should know how to select suitable dosage and injection system according to actual conditions.
In a preferred embodiment, the invention provides the herbal-composition for treating GI inflammatory disease in object, it comprises the extract available from Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica.
In a preferred embodiment of the invention, provide the method by treating GI inflammatory disease to the herbal-composition of subject effective amounts, described herbal-composition comprises the extract from Pogostemon cablin and perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica.
In in of this embodiment, described herbal-composition regulates the expression of pro-inflammatory cytokine in the object suffering from GI inflammatory disease, weaken the destruction of epithelial barrier or regulate gut function.Especially, the present invention relates to the herbal-composition of use medication amount to the Expression of Macrophages pro-inflammatory cytokine suppressing LPS to stimulate, the destruction to epithelial barrier of weakening IFN γ induction and adjustment gut function.These activity contribute to the beneficial effect of said preparation antagonism GI inflammatory disease or illness, described GI inflammatory disease or illness comprise inflammatory bowel disease (ulcer disease and Crohn disease), intestinal irritable syndrome, chylous diarrhea and opiate withdrawal syndrome, such as, suffer from diarrhoea.
In some embodiments of the present invention, above-mentioned often kind of herbal ingredients in the composition can be as follows: Pogostemon cablin is about 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47% to about 48%w/w, w/v or v/v; Perilla leaf is about 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15% to about 16%w/w, w/v or v/v; Dried orange peel is about 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31% and 32%w/w, w/v or v/v; The bighead atractylodes rhizome is about 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31% and 32%w/w, w/v or v/v, and the root of Dahurain angelica is about 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15% to about 16%w/w, w/v or v/v.
In one embodiment, as shown in table 1, often kind of herbal ingredients units is by weight as follows: Pogostemon cablin accounts for about 33 % by weight of composition; Perilla leaf account for composition about 11 % by weight, dried orange peel account for composition about 22 % by weight, the bighead atractylodes rhizome accounts for about 22 % by weight of composition, and the root of Dahurain angelica accounts for about 11 % by weight of composition.
Table 1
| No. | Title | Amount (g) | % by weight |
| 1 | Pogostemon cablin | 326.8 | 33 |
| 2 | Perilla leaf | 108.9 | 11 |
| 3 | Dried orange peel | 217.9 | 22 |
| 4 | The bighead atractylodes rhizome | 217.9 | 22 |
| 5 | The root of Dahurain angelica | 108.9 | 11 |
| Amount to | 980.4 | 100 |
In another embodiment, herbal-composition is the extraction of essential oil thing from herbal ingredients of medicine effective quantity, and described essential oil is by carrying out steam distillation to extract forming the aqueous solution to herbal ingredients.
Present invention also offers herbal-composition, it includes the above-mentioned composition of effective amount and pharmaceutically suitable carrier, diluent or excipient or its combination.
In in of this embodiment, above-mentioned composition is pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
In another embodiment, the herbal-composition being used for the treatment of GI inflammatory disease described in comprises the extract available from Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome, the root of Dahurain angelica, the bark of official magnolia, the tuber of pinellia, Poria cocos, Radix Glycyrrhizae, the shell of areca nut, balloonflower root, ginger and date.
In some embodiments of the present invention, above-mentioned often kind of herbal ingredients in the composition can be as follows: Pogostemon cablin is about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% to about 20%w/w, w/v or v/v; Perilla leaf is about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7% to about 7.5%w/w, w/v or v/v; Dried orange peel is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% to about 15%w/w, w/v or v/v; The bighead atractylodes rhizome is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% to about 15%w/w, w/v or v/v; The root of Dahurain angelica is about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7% to about 7.5%w/w, w/v or v/v; The bark of official magnolia is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% to about 15%w/w, w/v or v/v; The tuber of pinellia is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% to about 15%w/w, w/v or v/v; Poria cocos is about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7% to about 7.5%w/w, w/v or v/v; Radix Glycyrrhizae is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% to about 15%w/w, w/v or v/v; The shell of areca nut is about 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7% to about 7.5%w/w, w/v or v/v; Balloonflower root is about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14% to about 15%w/w, w/v or v/v; Ginger is about 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4% to about 2.5%w/w, w/v or v/v; And date is about 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4% to about 3.5%w/w, w/v or v/v.
In the another kind of this embodiment is implemented, as shown in table 2, the percentage by weight of often kind of herbal ingredients is as follows: Pogostemon cablin accounts for about 15 % by weight of composition; Perilla leaf accounts for about 5 % by weight of composition; Dried orange peel accounts for about 10 % by weight of composition; The bighead atractylodes rhizome accounts for about 10 % by weight of composition; The root of Dahurain angelica accounts for about 5 % by weight of composition; The bark of official magnolia accounts for about 10 % by weight of composition; The tuber of pinellia accounts for about 10 % by weight of composition; Poria cocos accounts for about 5 % by weight of composition; Radix Glycyrrhizae accounts for about 10 % by weight of composition; The shell of areca nut accounts for about 5 % by weight of composition; Balloonflower root accounts for about 10 % by weight of composition; Ginger accounts for about 1.5 % by weight of composition; And date accounts for about 2.5 % by weight of composition.
Table 2
| No. | Title | Amount (g) | % by weight |
| 1 | Pogostemon cablin | 326.8 | 15 |
| 2 | Perilla leaf | 108.9 | 5 |
| 3 | Dried orange peel | 217.9 | 10 |
| 4 | The bighead atractylodes rhizome | 217.9 | 10 |
| 5 | The root of Dahurain angelica | 108.9 | 5 |
| 6 | The bark of official magnolia | 217.9 | 10 |
| 7 | The tuber of pinellia | 217.9 | 10 |
| 8 | Poria cocos | 108.9 | 5 |
| 9 | Radix Glycyrrhizae | 217.9 | 10 |
| 10 | The shell of areca nut | 108.9 | 5 |
| 11 | Balloonflower root | 217.9 | 10 |
| 12 | Ginger | 32.7 | 1.5 |
| 13 | Date | 54.5 | 2.5 |
| Amount to | 2157 | 100 |
In a further preferred embodiment, present invention also offers the method for treating GI inflammatory disease in object, it comprises the herbal-composition to described subject effective amounts, and described herbal-composition comprises the extract from Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome, the root of Dahurain angelica, the bark of official magnolia, the tuber of pinellia, Poria cocos, Radix Glycyrrhizae, the shell of areca nut, balloonflower root, ginger and date.
In in of this embodiment, described GI inflammatory disease is selected from: intestinal disease, comprises ulcer disease and Crohn disease, IBS, chylous diarrhea, opiate withdrawal syndrome and diarrhoea.Described herbal-composition regulates the expression of pro-inflammatory cytokine in the object suffering from GI inflammatory disease, weaken the destruction of epithelial barrier or regulate gut function.
These and other aspects, features and advantages of the present invention are understood by referring to detailed description herein, and the multiple key element by particularly pointing out in the dependent claims and combination are realized.Should be understood that foregoing general of the present invention describes with detailed description is hereafter exemplary and explanatory preferred embodiment of the present invention, but not restricts the present invention, as claimed.Some embodiments are hereafter portentous, although mention in an embodiment and include numerical value, result and/or data.In addition, following examples can comprise the data compilation of the data that the multiple times of representative in exploitation related to the present invention and experimentation collect.
Embodiment 1. ageratum (HXZQ) essential oil
In another preferred embodiment of the present invention, essential oil is processed: in order to prepare the prescription of herb formulation by following steps, crude product medicinal material Pogostemon cablin, the bighead atractylodes rhizome (frying), dried orange peel, perilla leaf and the root of Dahurain angelica are weighed, these crude product medicinal materials are put into the extraction vessel of volatile oil, add after a small amount of water soaked, add again and add water to submerge these medicinal materials, within 8 hours, heat by steam distillation and extract volatile oil, distillate is placed in separatory funnel, leave standstill, be then separated volatile oil and remaining distillate on upper strata.In addition, in the residue volatile oil of cleaning, 95% ethanol is added.Finally, this mixture of volatile oil is preserved in sealing.Also medicinal material residue is preserved.
In the preferred embodiment, the prescription (weight of gross weight and often kind of herbal ingredients is as shown in table 1) of a herb formulation can produce about 1ml volatile oil.The recovery rate of volatile oil is 1.0%.
Embodiment 2 HXZQ extract
In this preferred embodiment of the present invention, the extract of herb formulation is prepared: the bark of official magnolia is placed in Multifunction hand extracting container according to following steps, with 60% alcohol extract three times, (first time is with the ethanol of 6 volumes, second time and the ethanol of third time with 4 volumes), each 1 hour, filter, merging filtrate, reclaim alcohol by decompression from filtrate, thus produce the pasty state extract I of clarification.
In the preferred embodiment, then the medicinal material residue of Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica extracted after ethereal oil is got, and these medicinal material residues and the tuber of pinellia (system), balloonflower root, Poria cocos, Radix Glycyrrhizae, the shell of areca nut, ginger and date are merged in Multifunction hand extracting container, add extracting in water twice: first time extracts 2 hours after interpolation 8 volume water; Second time is extracted 1 hour after interpolation 6 volume water; Filter; Merging filtrate and the distillate as the residue extracted after volatile oil; Reducing pressure and being concentrated into relative density is 1.12 (80 DEG C), thus produces the pasty state extract of clarification, the pasty state extract II of centrifugal acquisition clarification.
Next step continues the pasty state extract I of concentrating clarifying and II is 1.35 (80 DEG C) to relative density, and dry in a vacuum, grinding, to produce extract, then seals preservation.
In the preferred embodiment, a herb formulation (weight of gross weight and often kind of herbal ingredients is as shown in table 2) can be produced the extract of 1ml volatile oil and about 0.54kg merging by ethanol and water, the ratio in extract shared by volatile oil is about 0.185%.
Embodiment 3.HXZQ soft capsule
The ageratum composition with advantageous characteristic is prepared: Pogostemon cablin, perilla leaf, the root of Dahurain angelica, the bighead atractylodes rhizome (frying), dried orange peel, the tuber of pinellia (system), the bark of official magnolia (ginger system), Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger and date by obtaining 13 kinds of medicinal materials.In a preferred embodiment of the invention, filter by the 60% alcohol extract bark of official magnolia 3 times.Merging filtrate is also condensed into pasty state extract I.The volatile oil of Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica is extracted to obtain the aqueous solution by steam distillation.Filtering solution after distillation, is collected in filtrate in another container.The medicinal material residue leached is decocted twice together with date with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger in water.Thing will be decocted and the above-mentioned aqueous solution merges.Filter, concentrated filtrate is to suitable volume, centrifugal and further supernatant concentration is become pasty state extract II.Merge pasty state extract I and II and be condensed into sticky extract (or dry in a vacuum).Grind this dry extracts and be sieved into fine powder.In the preferred embodiment, in ageratum extract, add volatile oil, refined soybean oil, beeswax and polysorbate80, in colloid mill, grind said preparation, be then compressed into soft capsule.
The anti-inflammatory activity of the Chinese herbal medicine formula ageratum that embodiment 4. is traditional and gut function regulate active
The feature of GI inflammatory disease or illness is that the cyclical level of pro-inflammatory cytokine raises usually, and macrophage is the prime producer of these pro-inflammatory cytokines.In order to determine whether ageratum extract has anti-inflammatory activity, with the macrophage of LPS process derived from bone marrow under the condition of presence or absence wrinkled glanthyssop vital energy preparation.
Materials and methods
The macrophage (BMDM) deriving from mouse bone marrow cells is utilized to study the anti-inflammatory activity of wrinkled glanthyssop vital energy preparation.QPCR is adopted to measure the mrna expression of cell factor.TRASNWELL plate is cultivated T84 cell.These cells form the individual layer of fully differentiation, for studying the impact of wrinkled glanthyssop vital energy preparation on epithelial barrier function in the first example, and in the second example, study wrinkled glanthyssop vital energy preparation add the impact of essential oil on epithelial barrier function.
In addition, intestinal smooth muscle bar is placed in organ bath, and mucous membrane is placed in You Si room to determine that wrinkled glanthyssop vital energy preparation regulates the effect of smooth muscle contraction and epithelial secretion.In this experiment, jejunal segment is longitudinally suspended from organ bath.The smooth muscle contraction of the electrical field stimulation in response to acetylcholine (modal cholinergic neurotransmitter), 5-HT or analog neuron stimulation is determined under the condition of presence or absence wrinkled glanthyssop vital energy preparation.The amplitude of smooth muscle Spontaneous Contraction is measured.
Mouse.C57BL/6 mouse is purchased from The Jackson Laboratories (Bar Harbor, ME).All experiments are all according to Institute of Laboratory Animal Resources, National Research Council, the principle that the animal used as test of Health and Human Services Publication is nursed and provide in guide for use (NIH 85-23, revision in 1996) is carried out.
The preparation of HXZQ extract and essential oil is see embodiment 1 to 3.In addition, in vitro study, Detoxi-Gel endotoxin removal gel (Thermo Scientific, Rockford, IL) is used to process herb extracts or essential oil to remove possible contaminative LPS.
The macrophage (BMDM) of derived from bone marrow and the preparation of cell culture.By the marrow from shin bone and femur being rushed in HyClone MEM α culture medium (Thermo) that (flush) enter containing 10%FBS to obtain non-adherent myelomonocyte from mouse.At 37 DEG C and 5%CO
2under, in HyClone MEM α culture medium, cultured cell spends the night, to exhaust adherent stroma cell.Collect non-adherent monocyte, with erythrocyte lysing buffer process to remove red blood cell.Then under the existence of 20ng/ml recombined small-mouse M-CSF (R & D Systems, Minneapolis, MN), cultivate these cells 7 days, to produce the macrophage broken up completely, culture medium is changed once every other day.Macrophage is laid in 6 orifice plates, and under presence or absence inflammatory mediator LPS, IFN or the condition of both, processes herb extracts or the essential oil of HXZQ, the same form 3 parts.After 24 hours, by cell harvesting in Trizol for separating of RNA, collect simultaneously not celliferous supernatant for by Luminex Assay measure cytokine release.Based on preliminary experiment, we have selected and have tested most suitable LPS concentration for individuality, as follows: be 1ng/ml for the mRNA expression of cytokines of BMDM, are 10ng/ml for the cytokine release of BMDM.
External smooth muscle contraction in organ bath.External smooth muscle contraction (Zhao etc., 2003) is measured according to previously described.Determine that smooth muscle is to the response of the electrical field stimulation (EFS) of acetylcholine, cholinergic neurotransmitter and analog neuron stimulation and Spontaneous Contraction amplitude.Tension force is represented as the tension force (Zhao etc., 2001) of unit cross-sectional area.
Epithelial cells in vitro ion transport in You Si room (Ussing chamber).According to previously described (Shea-Donohue etc., 2001), small intestine was placed in You Si room without muscle section.Measure short circuit current (short-circuit current, I
sc) in response to adding acetylcholine or the concentration dependent change to mucous membrane side interpolation glucose to serosa side.Do average, to obtain the average response of every animal to the response of all tissue segments of individual animals to acetylcholine or glucose.
The Microsnap hole of transepithelial electrical resistance (trans-epithelial electrical resistance, TEER) measures.T84 cell (CCL-248, ATCC) (a kind of people's colon epithelial cell system) is cultivated in the DMEM/F12 culture medium containing 5%FBS.In the every hole 5 × 10 of the upper inoculation of 12 hole transwell plates (Costar 3460, Corning Incorporated) with 12mm insert
5individual cell, at 37 DEG C and 5%CO
2under hatch.Add 1.5ml culture medium in the bottom (Basolateral) in hole, 0.5ml culture medium is added on room, top (top side).Cell converges after 10 to 14 days in cultivation and is polarized to epithelia monolayers.Then, under the condition of presence or absence IFN-γ (adding Basolateral to, a kind of known epithelial barrier disrupting agent), with herb extracts or this individual layer of essential oil process of adding top side and basolateral HXZQ to.Monitor TEER every day before treatment, then within 24 hours, 48 hours, again monitor after treatment.
RNA extracts, cDNA synthesizes and real-time quantitative PCR.According to previously described (Zhao etc., 2010), from cultured cell, extracted total serum IgE.Adopt the first chain cDNA synthase kit (MBI Fermentas, Hanover, MD), with six aggressiveness random primers, RNA sample (2mg) reverse transcription is become cDNA.As described in, carry out real-time quantitative PCR (qPCR) by iCycler detection system (Bio-Rad, Hercules, CA).After doing normalization relative to 18S rRNA, the multiple change of target gene mrna expression is relative to each supporting agent group mouse.The primer of qPCR is designed by Beacon Designer 7.0 (Premier Biosoft International, Palo Alto, CA) and is synthesized by the biopolymer laboratory (Baltimore, MD) of Sigma or University of Maryland.
Data analysis.Use one-way analysis of variance to carry out statistical analysis by Newman-Keuls test to compare response and gene expression.Suitable supporting agent contrast, time contrast and age matched control are also included within whole research.
Results and discussions
HXZQ extract or essential oil suppress to express/produce pro-inflammatory cytokine in macrophage consumingly
As one of the Major Members of intestines immunity, be present in intestinal lamina propria (Zhao A. etc., 2008 macrophage composition; Weber B etc., 2009; Platt AM etc., 2008).Under normal operation, gut macrophages shortage reduces the response of inflammatory stimulus or this response.During disease incidence, due to raising of circulating monocytic cell, in lamina propria, the number of macrophage improves.These macrophages raised discharge a large amount of pro-inflammatory cytokines, such as IL-1 β, L-12, TNF α and IFN γ, these cell factors inflammation generation and development both in be all important (Weber B etc., 2009).Finally, macrophage affects differentiation and amplification (Weber B etc., 2009 of Th1/Th17 inflammatory effector cell by release IL-12 and IFN γ; McKeernan DP etc., 2009).In view of IBD to raise as feature with pro-inflammatory cytokine cyclical level and macrophage is that this is true for the prime producer of these pro-inflammatory cytokines, although many Chinese herbal medicines all have anti-inflammatory property (Yang Z etc., 2012), we still determine to determine whether HXZQ can suppress macrophage to produce pro-inflammatory cytokine.We are to being generally considered to be pure full-brown macrophage group and being usually studied as the macrophage (BMDM) of the derived from bone marrow of the valuable cell model (Zhang X etc., 2001) of assessment anti-inflammatory activities.The HXZQ extract of three kinds of variable concentrations is tested: some major proinflammatory cytokine up regulation that the HXZQ (10,50 and 100 μ g/ml) of the three kinds of variable concentrations tested suppresses LPS to stimulate all consumingly, comprise IL-12p40, IFN γ, IL-1 β, IL-6 and TNF α (Figure 1A-E).
Carry out subsequent experimental to determine the effect of the essential oil from HXZQ.Similar with extract, concentration is that IL-12p40, IFN γ, IL-1 β, IL-6 and TNF α that the essential oil of 1,5 and 20 μ l/ml significantly suppresses LPS (10ng/ml) to stimulate raises (Fig. 2 A-E).An independent experiment shows further, and the oil being low to moderate 0.2 μ l/ml can suppress the rise (Fig. 2 F) of all subject cell factors.These Data supports HXZQ herb formulation has strong anti-inflammatory property.
HXZQ or essential oil weaken/block the epithelial barrier dysfunction that IFN γ induces.
It is important for controlling Intestinal permeabiligy for host defense, this is because, the molecule that the permeability improved impels a large amount of intraluminal bacterium, antigen or other pathogen to produce is by Mucosa Barrier, and this brings out immune activation (Turner JR etc., 2009).The epithelial cell of mucous membrane surface forms individual layer barrier, and its restriction pathogen or product are exposed to lower floor's immunocyte, and therefore, maintain complete Mucosa Barrier is one of most important factor in immune homeostasis as can producing evidence to prove.In fact, in the patient suffering from IBD, observed enterocyte barrier dysfunction.Since determined the anti-inflammatory activity of HXZQ in macrophage, next whether we just can affect epithelial barrier function to HXZQ has been determined.We cultivate T84 cell (a kind of people's colon epithelial cell system), then process it with HXZQ extract or essential oil under the existence of IFN γ (a kind of known epithelial barrier disrupting agent).Measure transepithelial electrical resistance (TEER) as permeability index.Consistent with previous report, in process after 24 hours and 48 hours, IFN γ effectively reduces the TEER of T84 individual layer, shows that permeability improves (Fig. 3).Concentration is that the existence of the HXZQ of 50 μ g/ml does not act on, but the TEER that 500 μ g/ml prevent this IFN γ to induce reduces, and shows to maintain normal barrier function (Fig. 3 A & B).Constantly little 24, concentration is that the essential oil of 0.1 μ l/ml and 0.2 μ l/ml has blocked the T84 simple epithelium that IFN γ induces completely and destroys, but within 48 hours, loses this effect (Fig. 3 D & E) after treatment.HXZQ extract obtains further checking to this abated effect of epithelial barrier dysfunction by the experiment of the inflow of measuring cascade blue dextran, in this experiment, the extract of 500 μ g/ml reduces the glucan of being induced by IFN γ and flows into (Fig. 3 C).It should be noted that both extract and essential oil all have no effect (data are not shown) to baseline TEER.
HXZQ extract suppresses smooth muscle contraction and enteric epithelium secretion.
GI inflammatory disease to cause such as abdominal pain, diarrhoea or constipation the gut function of symptom abnormal relevant.Whether we regulate gut function to be studied to HXZQ extract.As shown, intestines are in response to acetylcholine (Fig. 4 A), electrical field stimulation (EFS, Fig. 4 B) smooth muscle contraction and intestines bar Spontaneous Contraction (Fig. 4 D) be subject to the remarkable suppression of 0.1mg/ml HXZQ extract, and suppressed by the HXZQ extract of 1mg/ml further or eliminate.After washing away extract, the response that great majority reduce can partially or completely recover (data are not shown).Finally, smooth muscle is eliminated (Fig. 4 C) to the response of 5-HT by 0.1mg/ml HXZQ extract.
The major physiological effect of enteric epithelium absorbs nutrients or ion, secretion fluid and maintains mucosal barrier function, therefore, also regulates the effect of enteric epithelium secretion to be studied to HXZQ.The HXZQ of 0.1mg/ml significantly suppresses the chloride ion secretion in response to acetylcholine, and under the condition of 1mg/ml, eliminate this secretion response (Fig. 5 A).5-HT is the important neurohormone mediator (Gershon MD etc., 1991) that central nervous system is unified in both peripheral nervous systems.Intestines be maximum 5-HT source (5-HT great majority from intestines addicted to chromium (enterochromaffin, EC) cell), and comprise a large amount of 5-HT receptor subtype (Gershon MD1991 participating in the Physiological effect of intestinal mucosa and smooth muscle function; Gershon MD 2004; Gershon MD 2003).5-HT also plays an important role in maintenance mucous membrane stable state.Change (Beaurepaire etc., 2009 of EC cell number and 5-HT content are disclosed in many intestinal diseases (comprising IBS, IBD and chylous diarrhea); Spiller R.2008).As expected, the HXZQ extract of 0.1mg/ml eliminates the response (Fig. 5 B) of enteric epithelium secretion to 5-HT.These data show, HXZQ can regulate gut function.
In a word, this research shows, HXZQ has strong anti-inflammatory activity and gut function regulates active.This herb extracts not only Immunosuppression cell produces pro-inflammatory cytokine, and the epithelial cell barriers blocking inflammatory mediator induction destroys.In addition, HXZQ can regulate intestinal smooth muscle and epithelium function.Therefore, HXZQ can be the potential therapeutic agent of the intestinal disease (such as IBD) that antagonism Th1 preponderates.
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Claims (32)
1. herbal-composition, it for treating gastroenteritis disease in object, and described herbal-composition comprises the extract available from Pogostemon cablin (Pogostemonis Herba) and perilla leaf (Perillae Folium), dried orange peel (Citri Reticulatae Pericarpium), the bighead atractylodes rhizome (Atractylodis Macrocephalae Rhizoma) and the root of Dahurain angelica (Angelicae Dahuricae Radix).
2. herbal-composition according to claim 1, wherein said Pogostemon cablin accounts for about 18 % by weight to about 48 % by weight of described composition; Described perilla leaf accounts for about 6 % by weight to about 16 % by weight of described composition; Dried orange peel accounts for about 12 % by weight to about 32 % by weight of described composition; The bighead atractylodes rhizome accounts for about 12 % by weight to about 32 % by weight of described composition, and the root of Dahurain angelica accounts for about 6 % by weight to about 16 % by weight of described composition.
3. herbal-composition according to claim 2, wherein said Pogostemon cablin accounts for about 28 % by weight to about 38 % by weight of described composition; Described perilla leaf accounts for about 9 % by weight to about 13 % by weight of described composition; Dried orange peel accounts for about 17 % by weight to about 27 % by weight of described composition; The bighead atractylodes rhizome accounts for about 17 % by weight to about 27 % by weight of described composition, and the root of Dahurain angelica accounts for about 9 % by weight to about 13 % by weight of described composition.
4. herbal-composition according to claim 3, wherein said Pogostemon cablin accounts for about 33 % by weight of described composition; Described perilla leaf accounts for about 11 % by weight of described composition; Dried orange peel accounts for about 22 % by weight of described composition; The bighead atractylodes rhizome accounts for about 22 % by weight of described composition, and the root of Dahurain angelica accounts for about 11 % by weight of described composition.
5. herbal-composition according to claim 1, wherein said extract is essential oil.
6. herbal-composition according to claim 5, wherein said essential oil extracts to form the aqueous solution by steam distillation.
7. herbal-composition according to claim 1, wherein said composition also comprises pharmaceutically suitable carrier, diluent or excipient, or its combination.
8. herbal-composition according to claim 7, wherein said composition is mixed with the form of pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
9. in the object suffering from gastroenteritis disease, treat the method for gastroenteritis disease, it comprises: to the herbal-composition comprising the extract from Pogostemon cablin and perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica of described subject effective amounts.
10. method according to claim 9, wherein said herbal-composition regulates the expression of pro-inflammatory cytokine, weakens the destruction of epithelial barrier or regulate gut function in the object suffering from gastroenteritis disease.
11. methods according to claim 9, wherein said composition also comprises pharmaceutically suitable carrier, diluent or excipient, or its combination.
12. methods according to claim 11, wherein said composition is formulated as into the form of pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
13. methods according to claim 9, wherein said gastroenteritis disease is selected from: intestinal disease, comprises ulcer disease and Crohn disease, IBS, chylous diarrhea, opiate withdrawal syndrome and diarrhoea.
14. herbal-compositions according to claim 1, it also comprises the extract from the bark of official magnolia (Magnoliae Officinalis Cortex), the tuber of pinellia (Pinelliae Rhizoma), Poria cocos (Poria), Radix Glycyrrhizae (Glycyrrhizae Radix et Rhizoma), the shell of areca nut (Arecae Pericarpium), balloonflower root (Platycodonis Radix), ginger (Zingiberis Rhizoma Recens) and date (Jujubae Fructus).
15. herbal-compositions according to claim 14, wherein said Pogostemon cablin accounts for about 10 % by weight to about 20 % by weight of described composition; Described perilla leaf accounts for about 2.5 % by weight to about 7.5 % by weight of described composition; Described dried orange peel accounts for about 5 % by weight to about 15 % by weight of described composition; The described bighead atractylodes rhizome accounts for about 5 % by weight to about 15 % by weight of described composition; The described root of Dahurain angelica accounts for about 2.5 % by weight to about 7.5 % by weight of described composition; The described bark of official magnolia accounts for about 5 % by weight to about 15 % by weight of described composition; The described tuber of pinellia accounts for about 5 % by weight to about 15 % by weight of described composition; Poria cocos accounts for about 2.5 % by weight to about 7.5 % by weight of described composition; Radix Glycyrrhizae accounts for about 5 % by weight to about 15 % by weight of described composition; The shell of areca nut accounts for about 2.5 % by weight to about 7.5 % by weight of described composition; Balloonflower root accounts for about 5 % by weight to about 15 % by weight of described composition; Ginger accounts for about 0.5 % by weight to about 2.5 % by weight of described composition; And date accounts for about 1.5 % by weight to about 3.5 % by weight of described composition.
16. herbal-compositions according to claim 15, wherein said Pogostemon cablin accounts for about 12.5 % by weight to about 17.5 % by weight of described composition; Described perilla leaf accounts for about 4 % by weight to about 6 % by weight of described composition; Described dried orange peel accounts for about 7.5 % by weight to about 12.5 % by weight of described composition; The described bighead atractylodes rhizome accounts for about 7.5 % by weight to about 12.5 % by weight of described composition; The described root of Dahurain angelica accounts for about 4 % by weight to about 6 % by weight of described composition; The described bark of official magnolia accounts for about 7.5 % by weight to about 12.5 % by weight of described composition; The tuber of pinellia accounts for about 7.5 % by weight to about 12.5 % by weight of described composition; Poria cocos accounts for about 4 % by weight to about 6 % by weight of described composition; Radix Glycyrrhizae accounts for about 7.5 % by weight to about 12.5 % by weight of described composition; The shell of areca nut accounts for about 4 % by weight to about 6 % by weight of described composition; Balloonflower root accounts for about 7.5 % by weight to about 12.5 % by weight of described composition; Ginger accounts for about 1 % by weight to about 2 % by weight of described composition; And date accounts for about 2 % by weight to about 3 % by weight of described composition.
17. herbal-compositions according to claim 16, wherein said Pogostemon cablin accounts for about 15 % by weight of described composition; Described perilla leaf accounts for about 5 % by weight of described composition; Described dried orange peel accounts for about 10 % by weight of described composition; The described bighead atractylodes rhizome accounts for about 10 % by weight of described composition; The described root of Dahurain angelica accounts for about 5 % by weight of described composition; The described bark of official magnolia accounts for about 10 % by weight of described composition; The tuber of pinellia accounts for about 10 % by weight of described composition; Poria cocos accounts for about 5 % by weight of described composition; Radix Glycyrrhizae accounts for about 10 % by weight of described composition; The shell of areca nut accounts for about 5 % by weight of described composition; Balloonflower root accounts for about 10 % by weight of described composition; Ginger accounts for about 1.5 % by weight of described composition; And date accounts for about 2.5 % by weight of described composition.
18. herbal-compositions according to claim 14, it also includes at least the first extract of effective amount and the mixture of the second extract, wherein
Described first extract comprises the purified alcoholic solution extract of the herbal medicine bark of official magnolia, and
Described second extract comprises medicinal material residue after the extraction of essential oil of the Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica that decoct together with date with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger combined with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger and the purified water solution extract of date.
19. herbal-compositions according to claim 18, wherein said first extract 60% alcohol extract three times, and filter to obtain filtrate.
20. herbal-compositions according to claim 19, wherein said filtrate merges, and reclaims, and be condensed into pasty state extract with ethanol.
21. herbal-compositions according to claim 18, wherein said essential oil extracts to form the aqueous solution from Pogostemon cablin, the bighead atractylodes rhizome (frying), dried orange peel, perilla leaf and the root of Dahurain angelica by steam distillation.
22. herbal-compositions according to claim 21, its Chinese medicine residue obtains after carrying out steam distillation to Pogostemon cablin, the bighead atractylodes rhizome (frying), dried orange peel, perilla leaf and the root of Dahurain angelica.
23. herbal-compositions according to claim 18, wherein said second extract is filtered, and filtrate is concentrated into suitable volume, centrifugal and be condensed into pasty state extract further.
24. herbal-compositions according to claim 18, mixture and the essential oil of wherein said at least the first extract and the second extract merge.
25. herbal-compositions according to claim 18, wherein said composition also comprises pharmaceutically suitable carrier, diluent or excipient, or its combination.
26. methods according to claim 25, wherein said composition is mixed with the form of pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
27. for the method for the treatment of gastroenteritis disease in the object suffering from gastroenteritis disease, it comprises: to the herbal-composition comprising the extract from Pogostemon cablin and perilla leaf, dried orange peel, the bighead atractylodes rhizome, the root of Dahurain angelica, the bark of official magnolia, the tuber of pinellia, Poria cocos, Radix Glycyrrhizae, the shell of areca nut, balloonflower root, ginger and date of described subject effective amounts.
28. methods according to claim 27, wherein said gastroenteritis disease is selected from: intestinal disease, comprises ulcer disease and Crohn disease, IBS, chylous diarrhea, opiate withdrawal syndrome and diarrhoea.
29. methods according to claim 27, wherein said herbal-composition regulates the expression of pro-inflammatory cytokine in the object suffering from gastroenteritis disease, weaken the destruction of epithelial barrier or regulate gut function.
30. methods according to claim 27, wherein said composition also comprises pharmaceutically suitable carrier, diluent or excipient, or its combination.
31. methods according to claim 30, wherein said composition is mixed with the form of pill, capsule, granule, tablet, supensoid agent, injection, syrup or tincture.
32. methods according to claim 27, the herbal-composition of wherein said effective dose also comprises the mixture of at least the first extract and the second extract, wherein
Described first extract comprises the purified alcoholic solution extract of the herbal medicine bark of official magnolia, and
Described second extract comprises medicinal material residue after the extraction of essential oil of the Pogostemon cablin, perilla leaf, dried orange peel, the bighead atractylodes rhizome and the root of Dahurain angelica that decoct together with date with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger combined with the tuber of pinellia, Poria cocos, balloonflower root, Radix Glycyrrhizae, the shell of areca nut, ginger and the purified water solution extract of date.
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| CN104922350A (en) * | 2015-04-29 | 2015-09-23 | 金先琦 | Traditional Chinese medicine for treating acute enteritis |
| CN110037294A (en) * | 2019-04-12 | 2019-07-23 | 吉林省九圣源生物科技有限公司 | A kind of herbal cuisine eaten for stomach and enteron aisle ulcer crowd |
| CN110038107A (en) * | 2018-01-14 | 2019-07-23 | 青岛市妇女儿童医院 | A kind of herb mixture for infantile chronic diarrhea |
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Also Published As
| Publication number | Publication date |
|---|---|
| SG10201706453XA (en) | 2017-09-28 |
| WO2013177067A2 (en) | 2013-11-28 |
| CN106511949A (en) | 2017-03-22 |
| SG11201407653SA (en) | 2014-12-30 |
| CN104363773B (en) | 2017-03-01 |
| WO2013177067A3 (en) | 2014-01-16 |
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