CN101869603B - Traditional Chinese medicine composite for protecting livers and preventing liver injury and preparation method thereof - Google Patents
Traditional Chinese medicine composite for protecting livers and preventing liver injury and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a traditional Chinese medicine composite for protecting livers and preventing liver injury and a preparation method and application thereof. The composite comprises the following bulk drugs in parts by weight: 20-30 parts of astraglus base, 10-15 parts of bighead atractylodes rhizome, 10-15 parts of tuckahoe, 10-15 parts of liquorice and 10-15 parts of Schisandra chinensis. The preparation method comprises the following steps of: adding water to the bulk drugs in percentage by weight and decocting, and concentrating until the concentration is equivalent to 0.55-0.75g.ml-1 which is the concentration of crude drugs. Proved by animal experiment pathological study, the spleen-tonifying and liver-protecting prescription of the invention does not have hepatotoxicity per se and has a certain function of preventing mice acute liver injury caused by CCl4. The prescription not only can relieve the liver injury of jasmine on normal mice but also can assist routine dosage of jasmine to prevent the mice acute liver injury caused by CCl4. The invention is safe and effective and has low price and is suitable for patients with chemical and medicament liver injury.
Description
Technical field
The present invention relates to the Chinese medicine composition field, particularly relate to Chinese medicine composition, its preparation method and the application of a kind of the liver protecting, prevention hepatic injury.
Background technology
Chinese medicine is that multiplying and being prosperous of the Chinese nation made huge contribution.In recent years, Chinese medicine more and more receives the attention of international medical educational circles as " green " " natural " medicine.Frequent along with international exchange, Chinese medicine health-product market and treatment field in overseas occupy a tiny space gradually." yet green " " natural " is not to be " safety ", and along with the extensive use of Chinese medicine, the toxic and side effects of some Chinese medicine also receives publicity day by day.
Several years ago receive Caulis Aristolochiae Manshuriensis in the Decoction of Gentiana for Purging the Liver-fire of extensive concern and be detected human kidney is had damage, also seized the finding of medicine such as Radix Aristolochiae, Fructus Aristolochiae has side effect to human body then, causes the attention of the medicinal medicine of people's centering.It is reported that the Chinese medicine that liver is had damage is except Radix Tripterygii Wilfordii, Herba Senecionis Scandentis, Fructus Toosendan, Rhizoma Dioscoreae Bulbiferae; Also have some Chinese medicines also to be considered to liver toxicity; For example Radix Polygoni Multiflori, Fructus Gardeniae, Fructus Xanthii, Rhizoma Atractylodis, Herba Menthae, Folium Artemisiae Argyi, Pericarpium Granati, Rhizoma Osmundae etc.; But mostly these toxicity are from clinician's impression, to get, and lack reliable clinical data and toxicological experiment and confirm.In above-mentioned Chinese medicine, some belongs to conventional Chinese medicine, and clinical practice is extensive, even still treats the medicine commonly used of hepatopathy.For example, Fructus Gardeniae utilization clinically from ancient times to the present is extensive and have remarkable curative effect, in the treatise on Febrile Diseases that doctor's sage-Zhang Zhongjing is shown, all has the compatibility of Fructus Gardeniae to use in the considerable prescription, and the application of You Yizai " piece of writing in the Treatise on Febrile Diseases TAI YANG syndrome " is the most extensive.Because the Fructus Gardeniae heat-clearing toxin-expelling functions is remarkable, have preferably transaminase lowering active with bilirubinic effect, modern Dr Akagi likes often it is used as key medicine facing diagnosis and treatment treatment acute, chronic hepatitis.
Because of in recent years many bibliographical informations being arranged, Fructus Gardeniae has liver toxicity, and heavy dose of the use can be caused liver injury.Bring puzzlement to the clinician thus, the side's of sending medication dare not be used Fructus Gardeniae, thereby influences clinical efficacy.Yet the Fructus Gardeniae consumption of the relevant hepatotoxic experimentation of Fructus Gardeniae exceeds tens of times of clinical usual amounts, greatly differs from each other with clinical practice dosage.Whether the Fructus Gardeniae of usual amounts can liver injury, does not still have the research report at present.Can both bring into play the clinical efficacy of Fructus Gardeniae through drug matching, and avoid the toxic and side effects of its liver injury again, existing literature also lacks report.Therefore; Need to observe the liver toxicity of the Fructus Gardeniae of clinical common dose, and it is compatible to protect the liver Chinese medicine with spleen invigorating to normal mouse and acute liver damage mice through zoopery; When observation was treated the acute liver damage model mice with Fructus Gardeniae, whether the liver toxicity of medicine can be avoided.
Summary of the invention
The technical problem that the present invention will solve provides Chinese medicine composition, its preparation method and the application of a kind of the liver protecting, prevention hepatic injury.Tcm treatment method theoretical direction through " knowing the liver disease will be transmitted into the spleen; tonifying the spleen in the ban "; According to hepatopathy using drugs with sour taste for invigoration in " Medical Treasures of the Golden Chamber ", help with burnt bitter, benefit with the medicine accent of sugariness principle; Select for use the Chinese medicine of 5 flavor medicine-food two-purposes such as the Radix Astragali, the Rhizoma Atractylodis Macrocephalae, Poria, Radix Glycyrrhizae, Fructus Schisandrae Chinensis to form compound recipe, be named the spleen invigorating side of protecting the liver.Confirm that through the research of science of zoopery hepatopathy this side itself does not have liver toxicity, and certain prevention CCl is arranged
4Due to the effect of chmice acute hepatic injury.This side can alleviate jasmine on normal mice, can assist the Fructus Gardeniae prophylactic treatment CCl of routine dose again
4Due to the chmice acute hepatic injury, therefore, characteristics such as it is safe, effective, inexpensive that the present invention has are fit to chemical and drug induced hepatic injury patient and take.
For solving the problems of the technologies described above; The Chinese medicine composition of a kind of the liver protecting of the present invention, prevention hepatic injury comprises that following bulk drugs processes: 20 parts~30 parts of Radix Astragali Preparatas, 10 parts~15 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts~15 parts in Poria, 10 parts~15 parts in Radix Glycyrrhizae, 10 parts~15 parts of Fructus Schisandrae Chinensis.
Preferred Chinese medicine composition bulk drugs is: 20 parts of Radix Astragali Preparatas, 10 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts in Poria, 10 parts in Radix Glycyrrhizae, 15 parts of Fructus Schisandrae Chinensis, this Chinese medicine composition is called " the spleen invigorating side of protecting the liver ".
In addition; Chinese medicine composition of the present invention also can make up with Fructus Gardeniae, and Chinese medicine composition promptly of the present invention can also be processed by following bulk drugs: 20 parts~30 parts of Radix Astragali Preparatas, 10 parts~15 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts~15 parts in Poria, 10 parts~15 parts in Radix Glycyrrhizae, 10 parts~15 parts of Fructus Schisandrae Chinensis, 9 parts~18 parts of Fructus Gardeniaes.
Preferred Chinese medicine composition bulk drugs is: 20 parts of Radix Astragali Preparatas, 10 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts in Poria, 10 parts in Radix Glycyrrhizae, 15 parts of Fructus Schisandrae Chinensis, 18 parts of Fructus Gardeniaes.
The method for preparing of the Chinese medicine composition of the liver protecting of the present invention, prevention hepatic injury comprises:
With said bulk drugs decocte with water, be concentrated into concentration and be equivalent to crude drug 0.55~0.75gml
-1
The principal indication that Chinese medicine composition of the present invention can be used for treating is the prevention hepatic injury, and therefore, said composition can be used for preparation prevention liver injury medicament.
Chinese medicine composition of the present invention can adopt the conventional method of Chinese medicine preparation to be prepared into the oral preparations (like granule, tablet, capsule or oral liquid) of different dosage form; Its consumption and the course of treatment can be done suitably adjustment according to the light and heavy degree of dosage form, patient's age, disease; Be generally every day dosage and be equivalent to this side's crude drug 60g; Can divide 3 times oral, with 1 month be a course of treatment, can use continuously 6 courses of treatment or the longer time.
Application of TCM rationale of the present invention combines modern medicine and scientific and technical means, and foundation " Medical Treasures of the Golden Chamber " cloud: " disease of husband liver, using drugs with sour taste for invigoration help with burnt bitter, and the medicine of using sweet drugs for tonifying is to transfer it ... ", liver being bold and firm viscera, the five elements belong to wood, are usefulness to dredge.Liver be disease, be prone to the native change of disease insult of gram spleen.Based on the idea of " not sick anti-biography the, disease is preapred for an unfavorable turn of events ", the treating the liver disease is tonifying the spleen in the ban, pacifies the spleen that it does not receive heresy, makes edema with the liver involved not invade spleen soil.The spleen being the foundation of acquired constitution, spleen invigorating also can be supported wood.
The present invention selects for use 5 flavor Chinese medicines such as the Radix Astragali, the Rhizoma Atractylodis Macrocephalae, Poria, Radix Glycyrrhizae, Fructus Schisandrae Chinensis to form compound recipe, is named the spleen invigorating side of protecting the liver.
The Radix Astragali is sweet in flavor and warm in property, in Shennong's Herbal, classifies as top gradely, has tonifying Qi and lifting yang, invigorating the spleen and benefiting QI, detoxifcation, liver protection effect, is monarch drug in the spleen invigorating side of protecting the liver.The main component of the Radix Astragali is glycoside, polysaccharide and trace element, clinical and modern pharmacology aspect, and have and protect the liver, diuresis, defying age, blood pressure lowering, heart tonifying, expansion coronary vasodilator, and effects such as whole body peripheral vessel, antibiotic, enhancing human body immunity power; Process be partial to walk in, can invigorating the spleen and replenishing QI, qi-supplementing, blood-engendering.Aspect zoopery, the Radix Astragali can improve serum IgG (immunoglobulin G) content of mice and the effect of humoral immunization, and the antibody systematic function of normal body is had obvious facilitation.Aspect clinical practice, be main compound recipe with the Radix Astragali, can make insufficiency of the spleen patient's IgG level rising, chronic hepatitis patient IgG rise to normal range by the preceding level decline of treatment, and immunoglobulin content is raise.With Radix Astragali oral liquid treatment chronic hepatitis the function of improving clinical symptoms and reducing SAIT (ALP) and effectively regulate immunity of organism can be arranged; On the other hand, can make the decline of chronic viral hepatitis B NK cytoactive and LTR (lymhocyte transformation rate) and the rising of SIL22R (soluble il-22 receptors), increase the weight of and obviously with the state of an illness; The Radix Astragali has the effect that improves NK cytoactive, LTR and reduction SIL22R to the chronic viral hepatitis B patients with mild.Prompting Radix Astragali ability enhanced NK cell is removed the ability of hepatitis B virus.
Rhizoma Atractylodis Macrocephalae nature and flavor sweetness and bitterness tepor, have spleen reinforcing, stomach reinforcing, dampness and in effect, and have hepatoprotective effect, in the spleen invigorating side of protecting the liver, be ministerial drug.Clinically in order to the treatment deficient QI of the spleen and stomach, have loose bowels, sweating, frequent fetal movement etc.Modern pharmacological research finds that the Rhizoma Atractylodis Macrocephalae contains compositions such as volatile oil, vitamin, contains atractylol, atractylone in its volatile oil, can increase gastrointestinal secretion and wriggling, the digestion of favourable the intestines and stomach, thereby ability the liver protecting, blood circulation promoting and blood sugar lowering; And ability ability nervous system regulation, normal food ability tonification human body, enhancing healthy energy.Even the Rhizoma Atractylodis Macrocephalae obviously improves the cellular immunization and the humoral immune function of body.Aspect zoopery; It is interior and external to the mouse spleen lymphocyte regulating action that Rhizoma Atractylodis polysaccharide PAM acts on body, and the result shows: Rhizoma Atractylodis polysaccharide PAM in finite concentration model state, can activate separately or collaborative ConA (concanavalin A)/PHA (phytohemagglutinin) promotes the normal mouse lymphocyte transformation and can obviously improve the excretory level of IL-2 (interleukin-2).PAM has restitution to the lymphocytic propagation function of immunosuppressed mice that hydrocortisone causes.Find also that simultaneously Rhizoma Atractylodis polysaccharide PAM is relevant with receptor, agonist isoproterenol to lymphocytic adjusting.Aspect the body defence, the Rhizoma Atractylodis Macrocephalae contains a large amount of T lymphocytes, bone-marrow-derived lymphocyte and macrophage.Because of immunologic hypofunction produces primed lymphocyte and antibody, the Rhizoma Atractylodis Macrocephalae can be removed materials such as senile cell and microorganism.Rhizoma Atractylodis Macrocephalae volatile oil can strengthen delayed hypersensitivity due to the mice DNCB (2, the 4-dinitrochlorobenzene), significantly improves Turnover of Mouse Peritoneal Macrophages EA rosette rate and can improve the activity of macrophage, enhancing body non-specific immunity, the growth of anticancer.
Poria is worked as medical material as food from ancient times, and Shennong's Herbal records it, classifies " top grade " as.The property slightly sweet flavor flat, go into the heart, spleen, lung meridian, have that spleen reinforcing is good for the stomach, an eliminating dampness and diuresis, hepatoprotective effect, and hormone-like effect and effect of reducing enzyme levels arranged, be adjuvant drug in the spleen invigorating side of protecting the liver.The Poria property of medicine is mild, mends and not high, sharp and not violent, can set upright, again can be eliminating evil, be preferable tonify deficiency diseases prevention key medicine.Modern pharmacology aspect, but pachyman enhances human body immunologic function; Pachyman has cold-resistant and antifatigue effect.Prove that at rat test Poria can improve the phagocytic function of macrophage, the anti-cancer tumor suppressor effect is arranged, and antibiotic, blood sugar lowering and effects such as reduction gastric acid, prevention gastric ulcer are arranged.Su-fuling can strengthen TNF (tumor necrosis factor) ability through BCG (bacillus calmette-guerin vaccine) activating macrophage generation in advance in the mice inside and outside; And have significant dose dependent to concern that it has the effect of obvious suppression property to the transplanted tumor cell in the mice body, the level of its inhibitory action and TNF is proportionate.
Radix Glycyrrhizae is also classified as top grade by Shennong's Herbal.Its nature and flavor are sweet flat, have invigorating the spleen and replenishing QI, heat-clearing and toxic substances removing, relieving cough and moistening lung.The gentle current twelve regular channels of these article property of medicine, coordinating the actions of various ingredients in a prescription so it is old to have another name called state, is messenger drug in the spleen invigorating side of protecting the liver.Modern pharmacological research finds that Radix Glycyrrhizae and glycyrrhizin are to CCl
4And hepatic injury and hepatocarcinoma due to chemical carcinogens methyl-ammonia diphenyl diimide have the significant protection effect.Glycyrrhizic acid has the effect that suppresses hepar damnification, and its enzyme effect of falling to chronic hepatitis is identified, and the digestive enzyme of glycyrrhizic acid can occur after acting on administration, and the transaminase reaches minimum behind 8hr; Infer that its mechanism of action is the direct protective action to liver plasma membrane.Further research proof, the mechanism of glycyrrhizic acid protection liver plasma membrane are to realize through the activity performance antiinflammatory action that suppresses phospholipase.To the effect of hepatitis virus, research confirms that glycyrrhizic acid has the effect that suppresses hepar damnification.The clinical application aspect, the glycyrrhizic acid diamidogen shows that through random pair treatment research its ALT (glutamate pyruvate transaminase) effect that reduces the chronic viral hepatitis patient is remarkable; Can take effect within the intravenously administrable 1w; Through the 2m treatment, the patient ALT more than 85% can recover normally obviously to be superior to the monoamine preparation; Serum SB (unconjugated bilirubin) also recovers normal synchronously after the medication, and can obviously reduce gamma Globulin in the later stage course of treatment.Showing that through liver biopsy control experiment research chronic type b, chronic hepatitis C patient all have the patient's liver histological about 80% to improve after the medication, mainly is inflammatory cell infiltration minimizing, hepatocellular degeneration, the downright bad minimizing.
Fructus Schisandrae Chinensis is unworthy of the honour top grade at Shennong's Herbal also name, be famous nourishing Chinese medicine.Because of its fruit sweet, sour, hot, bitter, salty bittersweet, so the name Fructus Schisandrae Chinensis.Have the YIN nourishing of astringing the lung, the arresting seminal emission that promotes the production of body fluid, QI invigorating reinforcing YIN-essence, support the five internal organs, effect of reducing enzyme levels, be the guiding drug in the spleen invigorating side of protecting the liver.The modern pharmacology prompting; Fructus Schisandrae Chinensis has regulating action to animal and human's nervous function; Can promote the generation of protein and hepatic glycogen, and the hepatic injury of chemical toxicant is shielded that its preparation can promote the hepatic glycogen heteroplasia; And can promote hepatic glycogen to decompose, the phosphorylation process of fructose and glucose is strengthened.Aspect zoopery, the oral schisandra chinensis ethyl hydrate extract of mice, behind the 3d, by the lumbar injection phenylalanine, finds that it infiltrates hepatic protein speed and significantly increases, and shows that protein synthesis increases continuously at every day 1 time.Rat injection CCl
4It is downright bad that the back produces lobules of liver; The activity of bsaophilic cmponent, mitochondrial DNA, glycogen, acid phosphatase, ATP enzyme, 5-nucleotidase, succinate dehydrogenase all reduces; Behind the Fructus Schisandrae Chinensis preparation for treating, the lobules of liver necrotic area dwindles, and basophilia, mitochondrion and glycogen increase; Various histochemistry enzymatic activitys increase, show the Fructus Schisandrae Chinensis ethanol extract to rat by CCl
4The high SGPT that causes (ALT, glutamate pyruvate transaminase) has tangible reduction effect.Give rabbit Fructus Schisandrae Chinensis ethanol extract in advance, can prevent CCl
4The mice SGPT that causes raises.The Fructus Schisandrae Chinensis polysaccharide has the better protect effect to the immune organ of tumor-bearing mice; And can suppress growth of tumor, and having the potential value of prevention and treatment cancer, it is in tumor-inhibiting action; Also spleen, thymus spinosity being increased sharply to give birth to acts on, and improves its body's immunological function thus.Aspect clinical application, when suffering from chronic hepatopathy, especially liver cirrhosis has alteration of intestinal flora in various degree often, and imbalance degree is often relevant with the liver function injury degree.Five tastes of medicinal herb can be adjusted intestinal microbial population effectively, and the prevention chronic hepatic injury shields to liver, is an ideal microecologic regulator and a hepatoprotective.Improving aspect the curative effect of medication, ancient medicine is thought to have only and Fructus Schisandrae Chinensis is smashed the back is used, and just can obtain five kinds of flavours that Fructus Schisandrae Chinensis includes, and five kinds of flavours are to guarantee that Fructus Schisandrae Chinensis can give full play to the basic of curative effect.Ming Dynasty Chen Jia plan is Sino-Japan its " book on Chinese herbal medicine covers an aromatic plant metioned in ancient books ": " Fructus Schisandrae Chinensis should be smashed the throwing of (then the five tastes are all) side in advance to pieces and fry in shallow oil." Qing Dynasty poplar scholar Thailand is more detailed to this argumentation in its " book on Chinese herbal medicine is stated hook unit ", think " the sweet acid of Fructus Schisandrae Chinensis skin and flesh, arduous in the nuclear, the five tastes are tool all, and the full person that distinguishes the flavor of is true.Be used as medicine and must smash nuclear, the square five tastes are equipped with ... ".This shows, must smash its nuclear (seed) when using Fructus Schisandrae Chinensis, can guarantee the curative effect of Fructus Schisandrae Chinensis.Li Shizhen (1518-1593 A.D.) is at its Compendium of Material Medica medium cloud: " Fructus Schisandrae Chinensis, acid is salty goes into lung and the kidney invigorating, hot bitter in taste acting on the heart and tonifying the lung are sweetly gone into Middle Palace and the strengthening the spleen stomach ".Be not difficult to find out that the curative effect of Fructus Schisandrae Chinensis is to embody through five kinds of contained flavours synergism that mutually combines, the five tastes all have its usefulness, lack one of which the curative effect of Fructus Schisandrae Chinensis is affected.Then be not difficult to fried its flavor if it is not smashed, so must smash to pieces into frying in shallow oil.
Take a broad view of the spleen invigorating side of protecting the liver, contain the meaning of decoction of four noble drugs side, embody the easy change of disease of hepatopathy, therefore pacify the spleen that does not receive heresy earlier, make spleen not taken advantage of by wooden heresy.Spleen invigorating protects the liver full side and has effects such as tonifying Qi and lifting yang, replenishing QI to invigorate the spleen, the liver protecting and ALT lowering.This side is used for normal mouse separately, does not all show activity and content of bilirubin effect that it has the rising transaminase, and this group mouse liver structure is normal, hepatocyte form Non Apparent Abnormality.This side and the same usefulness of high dose Fructus Gardeniae water extract prevented the hepatocellular degeneration that the high dose Fructus Gardeniae is caused effectively, reduced the transaminase activity that Fructus Gardeniae causes and the rising of content of bilirubin, shows the value of the spleen invigorating side of protecting the liver as Fructus Gardeniae attenuation compatibility prescription.
Chinese medicine composition of the present invention confirms to have the effect that alleviates hepatic injury through zoopery, has following advantage:
1) the spleen invigorating side of protecting the liver nature and flavor acid, bitterly, sweet possesses, closes, help with burnt bitter in " Medical Treasures of the Golden Chamber " hepatopathy using drugs with sour taste for invigoration, benefit with the medicine accent of sugariness principle;
2) moisten foster the foundation of acquired constitution to protect the liver through ridging, embodied the traditional Chinese medical science " knowing the liver disease will be transmitted into the spleen, tonifying the spleen in the ban " and " treatment of disease must aim at its pathogenesis's " the rule of treatment, " preventive treatment of disease " thought of treating the location first where the pathogens havn;
3) the spleen invigorating side of protecting the liver can alleviate jasmine on normal mice, can assist Fructus Gardeniae prophylactic treatment CCl again
4Due to the chmice acute hepatic injury;
4) the prescription Chinese medicine of the spleen invigorating side of protecting the liver is the article of medicine-food two-purpose, even Fructus Gardeniae also is put into " be food be again the article list of medicine ", can be by the relieved use of routine dose.
Description of drawings
Below in conjunction with the accompanying drawing and the specific embodiment the present invention is done further detailed explanation:
Fig. 1 is the pathological change figure that respectively organizes mouse liver (HE dyeing, * 400) among the embodiment 7, and wherein, 1-a is a normal group, the 1-b high dose group, and the high agent spleen invigorating of 1-c group, the 1-d spleen invigorating protects the liver group;
Fig. 2 is the variation diagram that the continuous modeling 6w among the embodiment 7 respectively organizes biochemical indicator, wherein, and among the 2-a, with the normal group comparison same period, * *: P<0.01, with the high dose Fructus Gardeniae water extract model group comparison same period, #:P<0.05; Among the 2-b, with the normal group comparison same period, *: P<0.05; Among the 2-c, with the normal group comparison same period, *: P<0.05; Among the 2-d, with the normal group comparison same period, * *: P<0.01, *: P<0.05;
Fig. 3 is that the pathological change of respectively organizing mouse liver and the spleen invigorating side of protecting the liver among the embodiment 8 is to CCl
4The prevention of hepatic injury is figure (HE dyeing, * 200) as a result, and 3-a is a normal group, and 3-b is a model group, and 3-c is middle dose groups, and 3-d is that spleen invigorating protects the liver group, and 3-e is that Fructus Gardeniae protects the liver group;
Fig. 4 is the variation diagram of respectively organizing biochemical indicator among the embodiment 8, wherein, among the 4-a, compares with normal group, and #:P<0.05, ##:P<0.01 are compared with model group in * *: P<0.01; Among the 4-b, compare with normal group, #:P<0.05, ##:P<0.01 are compared with model group in * *: P<0.01; Among the 4-c, compare with normal group, *: P<0.05, compare with model group * *: P<0.01, does not have diversity; Among the 4-d,, there is not diversity with normal group and the model group comparison same period.
The specific embodiment
Weighting raw materials: Radix Astragali Preparata 20g, Rhizoma Atractylodis Macrocephalae 10g, Poria 10g, Radix Glycyrrhizae 10g, Fructus Schisandrae Chinensis 10g, decocte with water 2 times can be concentrated into concentration and be equivalent to crude drug 0.55gml
-1
Weighting raw materials: Radix Astragali Preparata 25g, Rhizoma Atractylodis Macrocephalae 12.5g, Poria 12.5g, Radix Glycyrrhizae 12.5g, Fructus Schisandrae Chinensis 12.5g, decocte with water 2 times can be concentrated into concentration and be equivalent to crude drug 0.65gml
-1
Weighting raw materials: Radix Astragali Preparata 30g, Rhizoma Atractylodis Macrocephalae 15g, Poria 15g, Radix Glycyrrhizae 15g, Fructus Schisandrae Chinensis 15g, decocte with water 2 times can be concentrated into concentration and be equivalent to crude drug 0.75gml
-1
Weighting raw materials: Radix Astragali Preparata 20g, Rhizoma Atractylodis Macrocephalae 10g, Poria 10g, Radix Glycyrrhizae 10g, Fructus Schisandrae Chinensis 10g, Fructus Gardeniae 9g, decocte with water 2 times can be concentrated into concentration and be equivalent to crude drug 0.55gml
-1
Embodiment 5
Weighting raw materials: Radix Astragali Preparata 25g, Rhizoma Atractylodis Macrocephalae 12.5g, Poria 12.5g, Radix Glycyrrhizae 12.5g, Fructus Schisandrae Chinensis 12.5g, Fructus Gardeniae 13.5g, decocte with water 2 times can be concentrated into concentration and be equivalent to crude drug 0.65gml
-1
Embodiment 6
Weighting raw materials: Radix Astragali Preparata 30g, Rhizoma Atractylodis Macrocephalae 15g, Poria 15g, Radix Glycyrrhizae 15g, Fructus Schisandrae Chinensis 15g, Fructus Gardeniae 18g, decocte with water 2 times can be concentrated into concentration and be equivalent to crude drug 0.75gml
-1
7 pairs of hepatotoxic preventive effect of Fructus Gardeniae of embodiment
1. material
1.1 laboratory animal
60 of c57bl male mices, body weight 20 ± 2g, the SPF level is provided by Shanghai Univ. of Traditional Chinese Medicine's Experimental Animal Center, the animal quality certification number: SCKK (Shanghai) 2007-0009.
1.2 the preparation of experiment medicine
High dose Fructus Gardeniae water extract: get Fructus Gardeniae 200g, add proper amount of clear water, decocts 2 times, the medicinal liquid of 2 decoctions is condensed into high concentration water's extract, concentration 0.298gml
-1
0.65gml
-1The spleen invigorating side of protecting the liver method for making: extracting liquorice 42g, Rhizoma Atractylodis Macrocephalae 42g, Poria 42g, Radix Astragali Preparata 84g, Fructus Schisandrae Chinensis 63g adds suitable quantity of water and decocts 2 times, is concentrated into concentration and is equivalent to crude drug 0.65gml
-1
The high dose Fructus Gardeniae is closed the spleen invigorating side of protecting the liver: get Radix Astragali Preparata 60g, Rhizoma Atractylodis Macrocephalae 30g, Poria 30g, Radix Glycyrrhizae 30g, Fructus Schisandrae Chinensis 45g, Fructus Gardeniae 108g adds suitable quantity of water and decocts 2 times, is concentrated into concentration and is equivalent to crude drug 0.606gml
-1
The preparation of Chinese medicine extraction liquid is accomplished by heart assist in Shanghai Univ. of Traditional Chinese Medicine's scientific experiments.
1.3 key instrument and equipment
Automatic dehydration machine (ASP300), rotary microtome (RM2035), embedding machine (EG1160), waxdip machine (HI1210), roasting sheet machine (HI1220) are German Leica company.Cryogenic refrigerator (70 ℃), Forma scientific product.The chromatography cabinet, Revco company.Desk centrifuge (microfuge lite), the Beckman Company products.The AA-200 analytical balance, Densere instrument company product.Olympus BX40 optical microscope, Japanese Olympus company.Decoct medicinal herbs equipment is provided by science and technology experimental center, school.
2. method
2.1 zoopery
Mice is divided into normal group, high dose Fructus Gardeniae water extract group (hereinafter to be referred as high dose group), high dose Fructus Gardeniae at random closes the spleen invigorating side of protecting the liver group (hereinafter to be referred as high agent spleen invigorating group), the spleen invigorating side of protecting the liver group (be called for short spleen invigorating and protect the liver group), every group is 15.Normal group mouse stomach drinking water, all the other are respectively organized mice and irritate the stomach relative medicine every day, and irritating the stomach amount is 0.2ml10g
-1, 6w (week) puts to death behind the 6w and respectively organizes mice continuously.Every mice gives 2% (2g/100ml) pentobarbital sodium according to 0.02ml10g
-1Intraperitoneal injection of anesthesia, eyeball is got blood.Blood is splashed into 1.5ml EP pipe, and room temperature leaves standstill 2hr, and with 3000rpm, 15min is centrifugal.Get serum, place-70 ℃ of refrigerators subsequent use.The dislocation of mice vertebra is put to death, open the abdominal cavity, get the liver lobus dexter, cut 0.6 * 0.6 * 0.2mm
3Organize 2, put into 10% (percentage by volume) formaldehyde fixed liquid respectively.The residue liver organization is put into 1.5ml EP pipe, places-70 ℃ of refrigerators subsequent use.
2.2 observation index
2.2.1 liver organization pathological observation
With the dehydration respectively of fixed liver organization BIAO and BEN, waxdip, embedding, slice thick 4 μ m, do HE dyeing (hematoxylin-eosin staining), light microscopic is observed hepatic pathology down and is changed.
2.2.2 serum biochemistry is learned index and is detected
The active reitman-frankel method that adopts of ALT (glutamate pyruvate transaminase); The active reitman-frankel method that adopts of AST (glutamic oxaloacetic transaminase, GOT); Alb (albumin) content adopts the bromocresol green method; TBiL (STB) content adopts sodium benzoate-caffeine colorimetry, is detected by SHUGUANG HOSPITAL clinical laboratory automatic biochemistry analyzer.
2.3 statistics
Utilize the ANOVA program in the SPSS12.0 software to carry out one factor analysis of variance, and compare in twos with the LSD program.
3. result
3.1 respectively organize the mouse liver microexamination
The liver hepatic tissue clear in structure of normal group mice and the spleen invigorating side of protecting the liver group mice, the liver plate is radial arrangement, and the hepatocyte form is normal.Behind high dose Fructus Gardeniae water extract modeling 6w, model group murine liver tissue structure disturbance, the general swelling and degeneration of hepatocyte; Mostly be that serious water sample becomes even balloon appearance becomes; Few sample becomes the master with fat, and the individual samples hepatocellular degeneration is lighter, and sinus hepaticus is narrow; The hyperplasia of withered Fou Shi group, lymphocytic infiltration.It is lighter that the high dose Fructus Gardeniae of intervening with the spleen invigorating side of protecting the liver is simultaneously closed the hepatic tissue structural deterioration of the spleen invigorating side of protecting the liver group mice; Have 8 lobules of liver structures normal in 15 samples, all the other mouse liver cell degeneration are lighter, are mostly that the loose water sample of endochylema becomes; Only 1 example becomes the master with fat; Get involved hepatocyte below half, and multidigit is with in I, the narrow rare expansion (see figure 1) that is mostly of sinus hepaticus.
3.2 respectively organize the variation of mice biochemical indicator
ALT of high dose group mice and AST activity and TBiL content all obviously raise than normal group, have compared significant difference (P<0.01, P<0.05) for 2 groups.It is little that spleen invigorating protects the liver the variation of organizing these indexs, with the normal group no significant difference.The amplitude that these indexs of high agent spleen invigorating group raise is lower than high dose group, with compared with normal, and there was no significant difference (seeing table 1).
Table 1 6w respectively organizes the variation (mean ± S) of mice ALT and AST active and TBiL and Alb content
Annotate: compare *: P<0.05, * *: P<0.01 with normal group;
Compare #:P<0.05 with model group.
Can be known that by table 1 and Fig. 2 behind high dose Fructus Gardeniae water extract continuous irrigation stomach mice 6w, high dose group has tangible hepatic injury, damage becomes the master with water sample, and few sample becomes the master with fat, and sinus hepaticus is narrow.The structural deterioration of high agent spleen invigorating group hepatic tissue is lighter, has 8 lobules of liver structures normal in 15 samples, and all the other mouse liver cell degeneration are lighter, is mostly that the loose water sample of endochylema becomes, and the prompting spleen invigorating protects the liver the liver injury that can prevent high dose Fructus Gardeniae water extract to cause.Behind the spleen invigorating side of the protecting the liver continuous irrigation stomach mice 6w, mouse liver pathology and normal group are as good as, and explain that spleen invigorating protects the liver the burden that can not increase the weight of liver, can relievedly use.
The preventive effect of 8 pairs of carbon tetrachloride induced mice of embodiment acute liver damage
1. material
1.1 laboratory animal
48 of healthy SPF level c57bl male mices, body weight 20 ± 2g is provided by Shanghai Univ. of Traditional Chinese Medicine's Experimental Animal Center.The animal quality certification number: SCKK (Shanghai) 2007-0011.
1.2 experiment medicine
Carbon tetrachloride solution (Carbon tetrachloride, CCl
4), analytical pure, molecular weight 153.82, Shanghai Chemical Reagent Co., Ltd., Sinopharm Group, lot number: T20070323.Olive oil (Oliveoil), chemical pure, Shanghai Chemical Reagent Co., Ltd., Sinopharm Group, lot number: F20070307.
1.2.1 middle dosage Fructus Gardeniae water extract and the preparation of the spleen invigorating side of protecting the liver, the side of plan certainly
Middle dosage Fructus Gardeniae water extract: get 250ml high dose Fructus Gardeniae water extract (referring to embodiment 7), add the equivalent distilled water diluting, concentration Fructus Gardeniae water extract 0.149gml in being prepared into
-1
The spleen invigorating side of protecting the liver: extracting liquorice 42g, Rhizoma Atractylodis Macrocephalae 42g, Poria 42g, Radix Astragali Preparata 84g, Fructus Schisandrae Chinensis 63g adds suitable quantity of water and decocts 2 times, is concentrated into concentration and is equivalent to crude drug 0.65gml
-1
1.2.2 Fructus Gardeniae is closed the preparation of the spleen invigorating side of protecting the liver
Get Radix Astragali Preparata 20g, Rhizoma Atractylodis Macrocephalae 10g, Poria 10g, Radix Glycyrrhizae 10g, Fructus Schisandrae Chinensis 15g, Fructus Gardeniae 18g adds suitable quantity of water and decocts 2 times, is concentrated into concentration 0.55gml
-1Chinese medicine extraction is accomplished by heart assist in Shanghai Univ. of Traditional Chinese Medicine's scientific experiments.
1.3 key instrument and equipment
Automatic dehydration machine (ASP300), rotary microtome (RM2035), embedding machine (EG1160), waxdip machine (HI1210), roasting sheet machine (HI1220) are German Leica company.Cryogenic refrigerator (70 ℃), Forma scientific product.The chromatography cabinet, Revco company.Desk centrifuge (microfuge lite), the Beckman Company products.The AA-200 analytical balance, Densere instrument company product.Olympus BX40 optical microscope, Japanese Olympus company.Decoct medicinal herbs equipment is provided by science and technology experimental center, school.
2. method
2.1CCl
4The foundation of model
Mice is divided into normal group, CCl at random
4Group (hereinafter to be referred as model group), middle dosage Fructus Gardeniae water extract group (hereinafter to be referred as middle dose groups), spleen invigorating protect the liver group, Fructus Gardeniae is closed the spleen invigorating side of protecting the liver group (protecting the liver group hereinafter to be referred as Fructus Gardeniae).Except that normal group was 8 mices, all the other every group was 10.Normal group mouse stomach drinking water, all the other mices are all irritated stomach with relative medicine, irritate the stomach amount and are 0.2ml10g
-1, continuous 5d.6d removes the normal group mice, and all the other are respectively organized mice and give 0.5% (percentage by volume) CCl
4/ olive oil solution, 0.1ml10g
-1Lumbar injection, behind the lumbar injection, all mice fasting 17hr, but still give drinking water.Behind the 17hr, according to 0.02ml10g
-1Dosage, mice give 2% pentobarbital sodium intraperitoneal injection of anesthesia, and eyeball is got blood.Blood is splashed into 1.5ml EP pipe, and room temperature leaves standstill 2hr, and with the EP pipe that blood is housed, with 3000rpm, 15min is centrifugal.Extract serum, place-70 ℃ of refrigerators subsequent use.The dislocation of mice vertebra is put to death, open the abdominal cavity, get the liver lobus dexter, cut 0.6 * 0.6 * 0.2mm
3Organize 2, put into 10% formaldehyde fixed liquid respectively.Surplus liver organization is put into 1.5ml EP pipe, places-70 ℃ of refrigerators subsequent use.
2.2 observation index
2.2.1 liver organization pathological observation
With the dehydration respectively of fixed liver organization BIAO and BEN, waxdip, embedding, slice thick 4 μ m, HE dyeing, light microscopic is observed hepatic pathology down and is changed, and intends hepatocellular degeneration grade scale (seeing table 2) certainly.
Table 2 hepatocellular degeneration grade scale
2.2.2 biochemistry detection
The active reitman-frankel method that adopts of ALT; The active reitman-frankel method that adopts of AST; Alb content adopts the bromocresol green method; TBiL content adopts sodium benzoate-caffeine colorimetry, is detected by SHUGUANG HOSPITAL clinical laboratory automatic biochemistry analyzer.
2.3 statistics
Utilize the ANOVA program in the SPSS12.0 software to carry out one factor analysis of variance, and compare in twos with the LSD program.Semi-quantitative analysis adopts the Ridit method statistic.
3. result
3.1 Fructus Gardeniae water extract and the spleen invigorating side of protecting the liver are to the pathological observation of the therapeutical effect of acute liver damage mice
3.1.1 respectively organize the mice gross examination of skeletal muscle
Injection CCl
4Before respectively organize the mice outward appearance and all do not have obvious different with mobility.5d lumbar injection CCl
4After, except that normal group, each energy of organizing mice reduces.The mice energy of 6d model group is starkly lower than other each groups.
3.1.2 respectively organize the mouse liver perusal
Normal group mouse liver perusal color and luster is scarlet, matter is soft, clear-cut margin, and model group mouse liver edge is blunt slightly, and liver has swelling slightly.All the other are respectively organized liver and change not obvious substantially.
3.1.3 respectively organize the mouse liver microexamination
The result is as shown in Figure 3, and normal group Mouse Liver leaflet structure is normal, and the liver plate is radial arrangement, and the hepatocyte form is normal.Model group liver plate arrangement disorder, the general degeneration of hepatocyte, water sample becomes and is that master's fat becomes all obvious with vesicle property, and part of hepatocytes balloon appearance becomes, and degeneration hepatocyte multidigit is with III in liver acinus I band, and severe patient II is with hepatocyte that degeneration is also arranged.The point-like or necrosis of kitchen range shape and the cell infiltration that are dispersed in are arranged.Middle dose groups hepatocyte injury situation is light than model group, is the water sample change that vesicle property fat becomes or II is with of being with at I, and change of balloon appearance and necrocytosis are rare, and sinus hepaticus is narrow.Spleen invigorating protects the liver group liver damage situation: hepatocyte fat change person is many at the I band, and observed balloon appearance becomes hepatocyte seldom in the II band.Fructus Gardeniae protects the liver group hepatocellular steatosis and hepatocellular ballooning degeneration seldom, and the hepatic necrosis situation do not see, is the lighter of pathological change in 4 groups.
3.1.4 respectively organize mouse liver cell degeneration semi-quantitative analysis
Table 3 shows that the normal group mouse liver cell does not have degeneration.The degeneration of model group mouse liver cell is serious, is mostly the IV level, with normal group comparing difference remarkable (P<0.001).The Fructus Gardeniae water extract of middle dosage and spleen invigorating protect the liver can be to alleviate CCl
4The hepatocellular degeneration that causes, this hepatocyte deformation extent of 2 groups is low than model group, but significant difference is not obvious.Fructus Gardeniae protects the liver the hepatocellular degeneration of group obviously than model group light (P<0.01), and the routine number of hepatocyte I, the degeneration of II level occupies the majority, and does not have the sample of IV level degeneration.
Table 3 is respectively organized the hepatocellular degeneration degree
Annotate: compare * * *: P<0.001 with normal group;
Compare ##:P<0.01 with model group.
3.2 Fructus Gardeniae water extract and the spleen invigorating side of protecting the liver are to the observation of therapeutical effect and the hepatotoxic biochemical indicator of acute liver damage mice
By table 4 and shown in Figure 4, CCl
4After the injection, model group mice ALT and AST activity and TBiL content significantly raise, and with compared with normal significant difference (all P<0.01) are arranged.The ALT and the AST activity of middle dose groups also raise, but lower than model group, and significant difference (P<0.05, P<0.01) is arranged, and TBiL content raises similar with model group.Spleen invigorating protects the liver group ALT activity and TBiL content raises obviously; With normal group significant difference (all P<0.01) is arranged; The active rising amplitude of AST then protects the liver group mice ALT and AST activity and TBiL content than model group obviously low (P<0.05) Fructus Gardeniae and also raises; But obvious lower than model group, wherein transaminase activity has been compared significant difference (all P<0.01) with model group.Alb content is no significant difference between 4 groups.
Table 4 is respectively organized the variation (mean ± S) of mice ALT and AST active and TBiL and Alb content
Annotate: compare *: P<0.05, * *: P<0.01 with normal group;
Compare #:P<0.05, ##:P<0.01 with model group
CCl
4Cause the chmice acute hepatic injury after the shot, ALT and AST activity and TBiL content obviously raise, and hepatocellular degeneration is serious, model group and compared with normal significant difference (all P<0.01).The Fructus Gardeniae water extract of middle dosage can suppress CCl effectively
4The transaminase activity that causes raises; All than model group group obviously low (P<0.05, P<0.01), pathological observation shows that also this group hepatocellular degeneration is lighter than model group for middle dose groups ALT and AST activity; But significant difference is not obvious, shows that the hepatic injury of Fructus Gardeniae prevention chmice acute has certain drug effect.The spleen invigorating side of protecting the liver has and reduces the active effect of acute liver damage aminotransferase in mice, this group AST activity and model group compared significant difference (P<0.05), still falling the enzyme amplitude is slightly poorer than Fructus Gardeniae, pathological change is similar with middle dose groups.The same usefulness of the Fructus Gardeniae of this side and middle dosage, the amplitude of falling enzyme is maximum in 3 medication groups, and with model group significant difference remarkable (P<0.01), this group hepatocellular degeneration is the lightest in 3 medication groups, with model group significant difference (P<0.01) is arranged equally.The effect of the spleen invigorating side of protecting the liver possibly be embodied in two aspects, with synergism of Fructus Gardeniae or the liver toxicity of minimizing Fructus Gardeniae.Though present embodiment can't differentiate that the Fructus Gardeniae of dosage can effectively prevent CCl in can pointing out to these two effects
4Due to the chmice acute hepatic injury, the spleen invigorating side of protecting the liver and Fructus Gardeniae compatibility can make preventive effect better.
Claims (6)
1. the Chinese medicine composition of a liver protecting, prevention hepatic injury is characterized in that: processed by following bulk drugs: 20 parts~30 parts of Radix Astragali Preparatas, 10 parts~15 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts~15 parts in Poria, 10 parts~15 parts in Radix Glycyrrhizae, 10 parts~15 parts of Fructus Schisandrae Chinensis.
2. the Chinese medicine composition of the liver protecting as claimed in claim 1, prevention hepatic injury, it is characterized in that: the weight portion of the crude drug of described Chinese medicine composition is: 20 parts of Radix Astragali Preparatas, 10 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts in Poria, 10 parts in Radix Glycyrrhizae, 15 parts of Fructus Schisandrae Chinensis.
3. the Chinese medicine composition of the liver protecting as claimed in claim 1, prevention hepatic injury; It is characterized in that: described Chinese medicine composition, process by following bulk drugs: 20 parts~30 parts of Radix Astragali Preparatas, 10 parts~15 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts~15 parts in Poria, 10 parts~15 parts in Radix Glycyrrhizae, 10 parts~15 parts of Fructus Schisandrae Chinensis, 9 parts~18 parts of Fructus Gardeniaes.
4. the Chinese medicine composition of the liver protecting as claimed in claim 3, prevention hepatic injury; It is characterized in that: the weight portion of the crude drug of described Chinese medicine composition is: 20 parts of Radix Astragali Preparatas, 10 parts of the Rhizoma Atractylodis Macrocephalaes, 10 parts in Poria, 10 parts in Radix Glycyrrhizae, 15 parts of Fructus Schisandrae Chinensis, 18 parts of Fructus Gardeniaes.
5. like the method for preparing of Chinese medicine composition of each described the liver protecting of claim 1 to 4, prevention hepatic injury, comprising:
With said bulk drugs decocte with water, be concentrated into concentration and be equivalent to crude drug 0.55~0.75gml
-1
6. the Chinese medicine composition like each described the liver protecting of claim 1-4, prevention hepatic injury prevents the application in the liver injury medicament in preparation.
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