CN101028282B - Use of low-molecular weight phaeophytin polyose sulfate in preparation of medicine for treating kidney disease - Google Patents
Use of low-molecular weight phaeophytin polyose sulfate in preparation of medicine for treating kidney disease Download PDFInfo
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- CN101028282B CN101028282B CN2007100643133A CN200710064313A CN101028282B CN 101028282 B CN101028282 B CN 101028282B CN 2007100643133 A CN2007100643133 A CN 2007100643133A CN 200710064313 A CN200710064313 A CN 200710064313A CN 101028282 B CN101028282 B CN 101028282B
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- algal polysaccharide
- polysaccharide sulfate
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- FQXBMKZVLSACGS-UHFFFAOYSA-N n,n-dimethylmethanamine;hexadecane;hydrobromide Chemical compound Br.CN(C)C.CCCCCCCCCCCCCCCC FQXBMKZVLSACGS-UHFFFAOYSA-N 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
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- BPATYLCKLFEYHC-UHFFFAOYSA-N oxolead;hydrate Chemical compound O.[Pb]=O BPATYLCKLFEYHC-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/02—Algae
- A61K36/03—Phaeophycota or phaeophyta (brown algae), e.g. Fucus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/734—Alginic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Alternative & Traditional Medicine (AREA)
- Urology & Nephrology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Botany (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
An application of the low-molecular polyalgose sulfate in preparing the medicines for treating the nephrosis, including renal failure, nephritis, nephrotic syndrome, etc, is disclosed.
Description
Technical field
The present invention relates to the pharmaceutical applications of low-molecular-weight algal polysaccharide sulfate, particularly the purposes of low-molecular-weight algal polysaccharide sulfate aspect preparation treatment kidney disease medicine.
Background technology
Sulfated fucan is one type of sulfated polysaccharides, is present in the Brown algae, is at first extracted from the palmate Thallus Laminariae (Thallus Eckloniae) with diluted acid in 1913 by Kylin.Kylin isolates the L-fucose after with the extract hydrolysis; He is with this polysaccharide called after fucoidin; Existing generally naming according to the nomenclature principle of polysaccharide is that fucoidan, Chinese are fucoidin, fucoidin, fucosan, fucoidan, fucoidan or sulfated fucan.Now people have comparatively clearly the composition of sulfated fucan and understand, and it is one type of chemical composition and the unusual polysaccharide of complicacy of structure, is main with fucose and sulfate, along with the kind difference of algae also contains other compositions such as galactose, xylose, alduronic acid.Thallus Laminariae (Thallus Eckloniae) Fucoidan is made up of monosaccharide such as fucose, galactose, xylose, glucuronic acid, arabinose.With fucose and galactose is main, and fucose and galactose are probably at 3:1.
The sulfated fucan chemical constitution is very complicated, and its structure of the sulfated fucan that is separated in the different Brown algaes has very big-difference.Up to the present; Structural research to the sulfated fucan that derives from Fucus Vesiculosus (Fucus vesiculosus) and yellow tang (Ascophyllum nodosum) is maximum; The Fucus Vesiculosus sulfated fucan mainly connects with α (1 → 3) glycosidic bond, and sulphation mainly occurs in C
4The position.The multinomial research of yellow tang sulfated fucan all shown wherein have a large amount of α (1 → 3) and α (1 → 4) glycosidic bond.Also have the structure of several kinds of Brown algae sulfated fucan to be in the news in addition.Thallus Laminariae (Thallus Eckloniae) (Ecklonia kurome) sulfated fucan is mainly α (1 → 3) and connects, and sulphation is at C
4The position.The sulfated fucan main chain that derives from tap algae (Cladosiphon okamuranus) and Chorda filum (L.) Stackh. (Chorda filum) is the fucose of α (1 → 3), and sulphation is at C
4The position, and the two all has a spot of 2-0-acetylation.
Fucus Vesiculosus, yellow tang sulfated fucan structure
Thallus Laminariae (Thallus Eckloniae) sulfated fucan structure
Chorda filum (L.) Stackh. sulfated fucan structure
About the structure of Thallus Laminariae (Thallus Eckloniae) sulfated fucan, most research datas shows that the Thallus Laminariae (Thallus Eckloniae) sulfated fucan mainly is to form with the L-fucose that α-(1 → 3) connects, and sulphation occurs in C
4Or C
2The position, and part Study shows that the L-fucose that has part (1 → 2) connection is as side chain.With Chorda filum (L.) Stackh. sulfated fucan structure among the last figure similarity is arranged.But the part acetyl group is arranged in the Chorda filum (L.) Stackh..And the shared ratio of different substituents group is also different.Certainly also have monosaccharide such as galactose, xylose, rhamnose in the molecule, galactose possibly participated in the composition of main chain, and xylose, rhamnose etc. to be form with side chain exist.
Existing many pieces of documents disclose the method for preparing and the pharmaceutical applications thereof of sulfated fucan and low-molecular-weight algal polysaccharide sulfate.The clear 46-2248 of Japan Patent adopts hexadecane pyridinium chloride or hexadecane trimethyl ammonium bromide and sulfated fucan to be reacted into the quaternary amine complex; Utilize the dissolubility difference of this complex again to salt; With ethanol, methanol and ion exchange resin treatment; Purification is removed Algin, neutral polysaccharide and other impurity, and obtains the fucoidin sulphuric acid acid esters of comparison purification.CN1129109A then discloses the alkali condensation method by dried kelp soaking, filtration for several times, secondary ethanol extraction, washing with alcohol, fit adjustment PH scope etc.CN1344565A then discloses another kind of method for preparing, comprises steps such as pretreatment of raw material, temperature control stirring and leaching, centrifugal, concentrated, ethanol precipitation, dehydrated alcohol dehydration.CN1517356A then is mixed with aqueous solution with fucoidan, adds hydrogen peroxide, hypochlorous acid or nitrous acid and salt thereof, and with the heating of gained mixed solution, the membrane ultrafiltration with molecular cut off 3000-5000 obtains fucoidan oligosaccharide.CN1560086A then discloses a kind of method for preparing of high sulfate radical content fucoidan, with hot water or sour water lixiviate Brown algae, makes the extracting solution that contains Brown algae polysaccharide sulfuric ester; The percetage by weight that this extracting solution is concentrated into polysaccharide is 2-10%, transfers PH5-8, adds chitosan solution and stirs; Centrifugal or cross collecting precipitation; Deposition is used for 5-10 times of saline solution extracts 2-4 time, clear liquid is collected in centrifugal or filtration, with this clear liquid dialysis or ultrafiltration desalination.CN1616494A is a raw material with the natural seaweed sulfated polysaccharide, with adding ascorbic acid and hydrogen peroxide, control reaction temperature in the Sargassum sulphuric acid sugar juice; The constant temperature degradation time is 0.5-3hr; Dialysis or ultrafiltration again, concentrating under reduced pressure makes the low molecular weight seaweed polysaccharide sulfate product of 4-100KDa.In addition, CN1670028A, CN1392160A, CN1197674A also disclose methods such as adopting flocculation respectively and have prepared Sargassum polysaccharides.In the present invention, above disclosure of invention is all introduced this paper as a reference in full.
In addition, more than invention also disclose sulfated fucan and low-molecular-weight algal polysaccharide sulfate and had anticoagulation, improved immunity, antitumor, antibiotic and antiviral, blood sugar lowering, radioprotective, inhibition ascites tumor isoreactivity.CN1228961A discloses the purposes of sulfated fucan aspect the treatment renal failure, and CN1206591A then discloses the purposes of sulfated fucan aspect the treatment renal glomerular disease.But there is not document to show the purposes of low-molecular-weight algal polysaccharide sulfate aspect kidney disease as yet.
Summary of the invention
The applicant finds that after deliberation low-molecular-weight algal polysaccharide sulfate is compared with sulfated fucan, aspect the treatment kidney disease, has beyond thought good therapeutic effect.Low-molecular-weight algal polysaccharide sulfate among the present invention; Be meant sulfated polysaccharides or oligosaccharide class material that sulfated fucan is made through certain suitable manner (the degraded mode includes but not limited to acid degradation method, alkaline degradation method, enzymatic degradation method, mechanical degradation method, free-radical oxidation edman degradation Edman etc.) degraded; Its molecular weight sulfated fucan molecular weight of not degrading is low; Concrete molecular weight ranges can be 1000～200000; Preferably 5000～100000, be more preferably 8000～80000.Sulfated fucan can derive from the Thallus Laminariae (Thallus Eckloniae) of artificial cultivation, also can be wild Brown algae Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, Mus tail algae, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae), yellow tang or Fucus Vesiculosus.Preferably derive from Thallus Laminariae (Thallus Eckloniae).
The purpose of this invention is to provide the purposes of low-molecular-weight algal polysaccharide sulfate aspect the treatment kidney disease.Kidney disease wherein includes but not limited to renal failure, nephritis and nephrotic syndrome etc.Wherein said nephritis includes but not limited to common clinically acute (glomerule) nephritis, chronic (glomerule) nephritis, pyelonephritis, latent nephritis, Henoch Schonlein purpura nephritis (henoch Schonlein purpura nephritis), lupus erythematosus nephritis (lupus nephritis) etc.
On the other hand, the invention provides the pharmaceutical composition that contains low-molecular-weight algal polysaccharide sulfate.Comprise the low-molecular-weight algal polysaccharide sulfate and at least a acceptable accessories of treating effective dose in the said compositions.The administering mode of said composition can be but be not limited to through mode administrations such as intravenous injection, oral, muscle, subcutaneous, skin surface, internal rectum, local injections; Its dosage form can but to be not limited to be injection, lyophilized injectable powder, injectable microsphere, liposome, tablet, capsule, water preparation, powder, paste, spray, granule, soft capsule, drop pill, gel, paster, unguentum etc., wherein preferred tablet, capsule, injection and lyophilized injectable powder.Those skilled in the art can prepare required dosage form easily according to the common practise of prior art and formulation art.
In the compositions of the present invention, the content of low-molecular-weight algal polysaccharide sulfate >=50% is preferably >=70%, and more preferably >=90%, the best is >=95%.The content of sulfated fucan can be 1mg～1000mg in the unit formulation, preferred 10mg～800mg, and more preferably 20mg～500mg, 20mg～300mg most preferably, the best is 30mg～100mg.
Sulfated fucan of the present invention can extract and purification, classification by following mode:
1. extract
Sulfated fucan can water, diluted acid or calcium chloride solution extract; In extracting solution, add Lead oxide hydrate, aluminium hydroxide, ethanol or quaternary ammonium salts cationic surfactant then; Sulfated fucan is precipitated out; In order to reduce the stripping of pigment, protein etc., can handle frond with high concentration alcohols or formalin earlier before extracting.
In recent years also successively the someone adopt methods such as microwave extraction, ultrasonic extraction and the extraction of flocculating polymer deposition.
2 purification
The thick sulfated fucan of preparation can contain part water solublity Algin, protein, laminaran, pigment etc. usually, need be further purified, and purification process has following several kinds:
The ethanol reprecipitation: the English one that exports to the west (the English first-class that exports to the west, Japan Fisheries Society's will, 1982,48 (12): 1771) to the thick algal polysaccharide sulfate aqueous solution of hot water extracting at 0.05M MgCl
2When existing, remove water-soluble algin as impurity with 20% precipitation with alcohol.Wang Zuoyun, Zhao Xuewu (Wang Zuoyun. Zhao Xuewu. separating and purification of the fucoidan of Sargassum phyllocystum Tseng et Lu,Sargassum horneri (Turn.) C. Ag. (Fucus horneri (Turn.)C.Ag.,Spongocarpus horneri Kutz.), laminaran and Algin. aquatic product journal .1985; 9 (1): 71) in the research Sargassum phyllocystum Tseng et Lu,Sargassum horneri (Turn.) C. Ag. (Fucus horneri (Turn.)C.Ag.,Spongocarpus horneri Kutz.) during sulfated fucan, with the thick sulfated fucan that makes water-soluble after, successively with 4M CaCl
2Remove Algin with 30% ethanol precipitation, then with the settle out sulfated fucan of purification of 80% ethanol.
The quaternary ammonium salts sedimentation method: utilize cationic surfactant such as hexadecylpyridinium chloride (CPC) or cetyl trimethyl ammonium bromide (CTAB) to produce sedimentary character sulfated fucan is precipitated with polyelectrolyte.
In extraction and purge process, for ion and the small-molecule substance of removing in the solution generally all adopts the method for dialysing.Also the someone adopts the ultra-filtration and separation method to get rid of the less material of molecular weight.Sometimes for to remove laminaran and the protein that is mingled in the extracting solution, can take enzyme digestion.(Fleury N andLahaye M.Studies on by-products from the industrlal extration of alglnate2.Chemieal structure analysis of fucans from the leach-water.J ApplPhycol such as Feury; 1993,5:605-610) when the side-product of the French Algin industry of research, just adopt glucanase and alcalase to remove wherein laminaran and protein.Separate laminaran and sulfated fucan and can also adopt ion-exchange-resin process, because the former is electroneutral, and the latter is the polyanion form.
3 classifications
Because the sulfated fucan chemical constituent is quite complicated; Chromatograph and electrophoretic examinations to the thick sulfated fucan prepared generally all present inhomogeneity, so people progressively use stage division that miscellaneous polysaccharide is divided into different fractions to further investigate.Stage division commonly used has two kinds: a kind of is ethanol precipitation, promptly utilizes different concentration of alcohol to be settled out different fraction, and another kind is a chromatography, utilizes gel filtration chromatography and ion-exchange chromatography to carry out classification.Ion exchange chromatography can be divided into the different fraction of charge with polysaccharide, and gel-filtration chromatography then carries out classification with polysaccharide according to the molecular weight size.Can also adopt hyperfiltration technique that sulfated fucan is carried out classification.
Below thereby the method that sulfated fucan degraded prepares low-molecular-weight algal polysaccharide sulfate then can be adopted several kinds:
1. acid degradation is under acid condition, and acid solution can cause the fracture of glycosidic bond in the polysaccharide, makes polysaccharide be degraded to low molecule fragment.Concentration, the temperature and time of control acid can obtain the catabolite of different molecular weight size.The difficult control of polysaccharide degradation products molecular weight distribution, sulfate radical content changes greatly.
2. the alkaline hydrolysis method often causes the modification of acidic polysaccharose and coming off of sulfate radical under alkali condition, influences product activity, therefore is not suitable for SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS.
3. the enzymatic isolation method enzymatic isolation method is to utilize the specificity glycosidase to reach the purpose of degraded through a certain glycosidic bond in the cracking polysaccharide.The enzymatic degradation reaction is easy to control because of specificity, high efficiency and degradation condition and the process of its height, and no side reaction etc. come into one's own in the polysaccharide degraded gradually.But because the specificity of enzyme is strong, therefore do not have extensive applicability, and the enzyme production cycle is long, loses activity easily, cost is high.These shortcomings all make this method can't apply at present.
4. the mechanical degradation method comprises methods such as ultrasound wave and microwave.These two kinds of methods are because energy consumption is high, and the instrument and equipment conditional request is high, and quantity of sample handling is little, can't be applied to commercial production at present.Ultrasound wave is radiating result show, radiated time length no matter, and the depolymerization molecular weight has individual lower bound; And depolymerizing substance has the molecular weight distribution that is rather narrow,
5. the free-radical oxidation degraded is for example with the hydrogen peroxide degradation method, and the degree of products therefrom is higher, and cost is lower, has bigger using value.
In addition, the disclosed method for distilling of institute's referenced patents and also can at random be adopted among this paper by those skilled in the art by the product of these methods preparation.
In a specific embodiment of the present invention, low molecule sulfated fucan is to adopt following method for preparing preparation: Thallus Laminariae (Thallus Eckloniae) is pulverized the back with the formalin soaked overnight, add the distilled water boiling water extraction; Extracting solution helps with kieselguhr and filters filter; Filtrating is earlier with tap water flowing water dialysis one day, with distill water dialysis one day, dialysis solution concentrated then; Adding ethanol to concentration is 75% deposition, precipitates the dry thick sulfated fucan that gets.Bullion is heavy water-soluble, at 0.05mol/L MgCl
220% ethanol precipitation is removed the water solublity Algin under existing, and back 75% ethanol precipitation is dialysed, concentrated to filtrating, promptly obtains the sulfated fucan of purification after the drying.Get an amount of Thallus Laminariae (Thallus Eckloniae) sulfated fucan, be dissolved in the distilled water; In this solution, add an amount of ascorbic acid and hydrogen peroxide, mix homogeneously, at room temperature stirring reaction is dialysed and ultrafiltration to reactant liquor, and ultrafiltrate is carried out concentrating under reduced pressure, with the concentrated solution lyophilization.
Below through the specific embodiment the present invention is further specified.Here want to be pointed out that; Below the specific embodiment only be used for explaining the present invention; Those skilled in the art are understanding under the prerequisite of spirit of the present invention; Can carry out corresponding conversion to the present invention according to the prior art and the common practise in present technique field, these technical schemes all fall within the scope of the present invention.
The specific embodiment
The preparation of embodiment 1 sulfated fucan
Sargassum is pulverized the back with 3.7% formalin soaked overnight; Add the distilled water boiling water extraction then, extracting solution helps with kieselguhr and filters filter, and filtrating is earlier with tap water flowing water dialysis one day; Then with distill water dialysis one day; Dialysis solution is concentrated, and adding ethanol to concentration is 75% deposition, precipitates the dry thick sulfated fucan that gets.Bullion is heavy water-soluble, at 0.05mol/L MgCl
220% ethanol precipitation is removed the water solublity Algin under existing, and back 75% ethanol precipitation is dialysed, concentrated to filtrating, promptly obtains the sulfated fucan of purification after the drying.Prepare four kinds of Sargassum sulfated fucan according to the method described above, its chemical constituent is analyzed as shown in the table:
The preparation of low-molecular-weight algal polysaccharide sulfate
1. take by weighing 150g Thallus Laminariae (Thallus Eckloniae) sulfated fucan, be dissolved in that to be made into concentration in the 10L distilled water be 1.5% solution; In this solution, add ascorbic acid and hydrogen peroxide, make their concentration reach 35mmol/L, mix homogeneously respectively; At room temperature stirring reaction is 2 hours, after reaction finishes, reactant liquor is dialysed and ultrafiltration; Ultrafiltrate is carried out concentrating under reduced pressure, again with the concentrated solution lyophilization; Making molecular weight ranges is the low-molecular-weight algal polysaccharide sulfate A of 8000-10000Da; Detecting this product weight average molecular weight through efficient gel permeation chromatography (HPGPC) method is 9.8KD.Chemical constituent analysis result: rock algae amount 28.4%, sulfate radical content 28.7%.
2. take by weighing 150g Thallus Laminariae (Thallus Eckloniae) sulfated fucan, be dissolved in that to be made into concentration in the 10L distilled water be 1.5% solution; In this solution, add ascorbic acid and hydrogen peroxide, make their concentration reach 2mmol/L, mix homogeneously respectively; At room temperature stirring reaction is 2 hours, after reaction finishes, reactant liquor is dialysed and ultrafiltration; Ultrafiltrate is carried out concentrating under reduced pressure, again with the concentrated solution lyophilization; Making molecular weight ranges is the low-molecular-weight algal polysaccharide sulfate B of 50KD-80KD; The weight average molecular weight that detects this product through efficient gel permeation chromatography (HPGPC) method is 72.2KD.Chemical constituent analysis result: fucose content 28.1%, sulfate radical content 29.2%.
3. take by weighing 150g Thallus Laminariae (Thallus Eckloniae) sulfated fucan, be dissolved in that to be made into concentration in the 10L distilled water be 1.5% solution; In this solution, add ascorbic acid and hydrogen peroxide, make their concentration reach 10mmol/L, mix homogeneously respectively; At room temperature stirring reaction is 2 hours; After reaction finishes, reactant liquor is dialysed and ultrafiltration, remove remaining ascorbic acid and hydrogen peroxide; Ultrafiltrate is carried out concentrating under reduced pressure, again with the concentrated solution lyophilization; Make the low-molecular-weight algal polysaccharide sulfate C of molecular weight ranges 15KD-30KD; The weight average molecular weight that detects this product through efficient gel permeation chromatography (HPGPC) method is 27KD.Chemical constituent analysis result: rock algae amount 28.5%, sulfate radical content 28.2%.
The preparation of embodiment 2 low-molecular-weight algal polysaccharide sulfate injections
Get low-molecular-weight algal polysaccharide sulfate 50g, add water for injection 500ml, mannitol 50g transfers pH value to 7.0, packing, lyophilization.
The preparation of embodiment 3 low-molecular-weight algal polysaccharide sulfate tablets
Get low-molecular-weight algal polysaccharide sulfate 50g, add microcrystalline Cellulose, polyvinylpyrrolidone mixes, and adds suitable quantity of water, and the system soft material is granulated drying.Particle adds cross-linking sodium carboxymethyl cellulose, magnesium stearate, mixes, and tabletting, every contains low-molecular-weight algal polysaccharide sulfate 10-200mg.
Embodiment 4 low-molecular-weight algal polysaccharide sulfates (sample B) are to the experimental study of rat chronic renal failure
1. low-molecular-weight algal polysaccharide sulfate partly excises the influence of the rat chronic renal failure that causes to kidney:
Choose 180～250 105 of male rats of gram (measuring no albuminuria before the art), be divided into normal control group and modeling group (95) at random, modeling group rat with pentobarbital sodium 30mg/kg intraperitoneal anesthesia after; Routine disinfection skin, the back left-hand cutout exposes left kidney; Peel off renal fascia, excise upper and lower utmost point excess of the kidney matter rapidly, immediately with the gelfoam hemostasis by compression; Drip several thrombin solutions in case of necessity, the kidney that resets then finishes operation.One Zhou Houzai undergos surgery, and excises right kidney.Postoperative is conventional raises; Get blood in 14 weeks of postoperative (rat is in heaven to die 16) and the rats in normal control group eye socket vein corner of the eyes; Measure rat blood serum creatinine, blood urea nitrogen, total protein, albumin level with 42 type Beckman biochemistry analyzer; According to measuring the result; The modeling rat is carried out random packet; Every group 14～15 (rejecting the following person of serum creatinine 176 μ mol/L), Thallus Laminariae (Thallus Eckloniae) sulfated fucan (being designated hereinafter simply as " the sample 1 ") 200mg/kg that promptly is used to prepare low-molecular-weight algal polysaccharide sulfate among model control group, low-molecular-weight algal polysaccharide sulfate B200, two dose groups of 100mg/kg and the embodiment 1 organizes, dexamethasone acetate 0.07mg/kg positive control drug group.The rat that other gets not operation is the normal control group, totally six groups, and the beginning experimental therapy; Gastric infusion once a day, continuous 30 days, normal all the filling once a day with model control group obeyed with the volume distilled water; Behind drug treatment, respectively organized the rat etherization in 10,20,30 days, eye socket is got blood, separation of serum; Carry out the mensuration of above-mentioned biochemical indicator, experimental result adopts t value method to carry out statistical procedures, and the result sees table 1.After getting blood for the last time,, get kidney and be fixed in 10% formalin routine paraffin wax embedding sacrifice of animal; Section; HE dyeing, the general nephridial tissue form of om observation is measured two mutually perpendicular vertical footpaths of glomerule (having vascular pole or/and urinary pole) and transverse diameter; The glomerule of 10 maximums is surveyed in every part of BIAO and BEN choosing, obtains the average area of each glomerule area and glomerule.To the renal tubules of each pathological section and ID range degree carry out pathological grading (,+, ++, +++) keep the score.The result adopts t check and WMW non parametric method to carry out statistical procedures, and the result sees table 2.
Table 1 is the result show: each is organized and rat 14 weeks behind kidney part resection operation forms chronic renal failures, with the normal control group relatively, serum creatinine, blood urea nitrogen all are significance and increase (p < 0.001), serum albumin obviously descend (p < 0.001).At drug treatment during 30 days, model control serum creatinine, urea nitrogen levels still are stable and increase (p < 0.001), the albumin state (p < 0.01～0.001) that obviously descends; After the low-molecular-weight algal polysaccharide sulfate treatment; Serum creatinine descends gradually, and treatment is when reaching 20～30 days, creatinine and blood urea nitrogen obviously descend (with model control group comparison P 0.05,0.01 or 0.001) or downward trend is arranged; And with the dosage increase, effect strengthens: serum albumin and total protein are not had obvious influence.
Table 2 pathological examination shows: the chronic kidney hypofunction rat due to kidney partly excises; Kidney tunicle connective tissue thickens, most of glomerule compensatory hypertrophy, glomerule area and the apparent in view increase of normal control group (p 0.001); Indivedual glomerule are sclerosis or glass transparent degeneration; Renal tubules cloudy swelling, fat change even atrophy, necrosis, interstitial fibers hamartoplasia, cell infiltration is obvious.With model control group relatively, low-molecular-weight algal polysaccharide sulfate can obviously reduce glomerule area and the tubulopathy branch that becomes, and relevant with dosage, change also has the trend of improvement to the kidney ID.Positive control drug dexamethasone acetate 0.07mg/Kg is suitable with low-molecular-weight algal polysaccharide sulfate B 200mg/Kg dose groups action intensity.
2 low-molecular-weight algal polysaccharide sulfates (sample B) are to the influence of the freezing rat chronic renal failure that causes
Choose 200～250 88 of male rats of gram (measuring no albuminuria before the art), be divided into normal control group (10) and modeling group (78) at random, modeling group rat with penta cling to must appropriate sodium 30mg/Kg intraperitoneal anesthesia after; Routine disinfection skin, the back left-hand cutout exposes left kidney; Peel off renal fascia, with immerse in advance cryoprobe in the liquid nitrogen bottle (0.6 * 1.4 * 25cm), totally four parts before and after the upper and lower utmost point of kidney and the outside successively; Freezing 45 seconds of every place, the kidney that resets then is after 2 weeks that finished to perform the operation; The right kidney of capable again excision, postoperative is conventional raises.Rat is got blood in 6 weeks of postoperative (so far dead 17) and the rats in normal control group eye socket vein corner of the eyes; Separation of serum; Measure rat blood serum creatinine, blood urea nitrogen, total protein and albumin with 42 type Beckman biochemistry analyzer; According to measuring the result modeling group rat is carried out random packet (rejecting the following person of serum creatinine 176 μ mol/L); Every group 9～10, i.e. model control group, low-molecular-weight algal polysaccharide sulfate B 200, two dose groups of 100mg/kg and sample 1 (200mg/kg) group, dexamethasone acetate 0.07mg/kg positive control drug group.The rat of getting not operation is the normal control group, totally 6 groups, and the beginning experimental therapy; Once a day, gastric infusion, continuous 40 days; Model control group, normal control group are all irritated clothes once a day with the volume distilled water, behind drug treatment, respectively organize the rat etherization in 10,20,30,40 days, and eye socket is got blood; Separation of serum carries out the mensuration of above-mentioned biochemical indicator, and the result sees table 3-1 and 3-2.Experiment finishes, with the rat sacrificed by decapitation.Get kidney, be fixed in 10% formalin, FFPE, HE dyeing, om observation.Nephropathy reason observation index and experimental result statistical procedures all are same as kidney and partly excise rat.The result sees table 4.
Table 3-1 and 3-2 result show, each is organized and rat 6 weeks after cryosurgery forms chronic renal failures, and with the comparison of normal control group, serum creatinine, blood urea nitrogen all are significance and increase (p < 0.001), serum albumin obviously descend (p < 0.05～0.001).At drug treatment during 40 days, model control group serum creatinine, urea nitrogen levels still are stable and increase (p < 0.001), the albumin state (p < 0.05～0.01) that obviously descends.After the low-molecular-weight algal polysaccharide sulfate treatment, serum creatinine descends gradually, when treatment reaches 30～40 days, creatinine, blood urea nitrogen obviously descend (with model control group relatively P 0.01 or 0.05) or downward trend is arranged, and increase with dosage, effect strengthens; Serum albumin and total protein there is not obvious influence.
Table 4 kidney pathological examination shows: the chronic kidney hypofunction rat pathological examination due to freezing is same as the chronic kidney hypofunction rat of kidney due to partly excising basically; Showing as kidney tunicle connective tissue thickens; Most of glomerule compensatory hypertrophy, glomerule area and the apparent in view increase of normal control group (p 0.001), indivedual glomerule are sclerosis or hyaloid hyaline degeneration; Renal tubules cloudy swelling, fat become, even atrophy, necrosis; Interstitial fibers hamartoplasia, cell infiltration is obvious.After low-molecular-weight algal polysaccharide sulfate treatment, high dose group can make glomerule area obviously descend (p < 0.05); The tubulopathy branch that becomes obviously reduces (p < 0.05), and low dose group also is as above variation tendency; The kidney ID is become the trend that improvement is also arranged.Positive control drug dexamethasone acetate 0.07mg/>kg is suitable with low-molecular-weight algal polysaccharide sulfate B200mg/kg dose groups action intensity.
The influence of the rat chronic renal failure that 3 low-molecular-weight algal polysaccharide sulfates (sample B) cause adenine
Choose 60 of body weight 160～200 gram Wistar rats, be divided into normal control group and modeling group immediately, modeling group rat oral gavage gives adenine 365mg/kg/ day; Totally 16 days, preparation chronic kidney hypofunction model, normal control group and modeling group rat freely ingest and drink water; In the 17th day etherization; Get blood by the eye socket vein corner of the eyes, separation of serum is measured rat blood serum creatinine, blood urea nitrogen, total protein and albumin with 42 type Beckman biochemistry analyzer.According to measuring the result, the moulding rat is divided into model control group, low-molecular-weight algal polysaccharide sulfate B 200, two dose groups of 100mg/kg and sample 1 (200mg/kg) group, dexamethasone acetate 0.07mg/kg positive control drug group, 10 every group at random; And the beginning gastric infusion, model control group waits capacity 0.5%CMC, except that the normal control group; All the other each groups continue to irritate stomach and give adenine, and dosage was 200mg/kg/ day, in administration the 40th day; Each organizes the rat etherization; The eye socket vein corner of the eyes is got blood, and separation of serum carries out the mensuration of above-mentioned biochemical indicator.Experimental result adopts the t check to carry out statistical analysis, and the result lists table 5 in.Experiment finishes, with the rat sacrificed by decapitation.Get fresh kidney immediately, observe mode of appearance and change, be fixed in 10% formalin, FFPE, section, HE dyeing, om observation.
Table 5 is the result show: the model control group rat oral gavage gives adenine after 16 days (before the treatment) serum creatinine, blood urea nitrogen and the comparison of normal control group, all is significance and increases (p < 0.001), albumin downward trend.Irritate the increase of stomach number of days with adenine, serum creatinine, urea nitrogen levels are the rising of carrying out property, albumin (p < 0.01) (the treatment back) that obviously descend.(being that adenine was irritated stomach after 16 days) its serum creatinine, blood urea nitrogen value and the more obviously increase of normal control group before each administration group rat treatment (p 0.001); Albumin obviously descends (p < 0.05) or downward trend is arranged; But compare with model control group, through statistical procedures, there was no significant difference.Behind low-molecular-weight algal polysaccharide sulfate (200 and 100mg/>kg) and the positive drug control group drug treatment serum creatinine and blood urea nitrogen obviously descend (with model control group relatively p 0.01 or 0.05); Low-molecular-weight algal polysaccharide sulfate (200mg/kg) and positive control drug also make serum albumin obviously rise, and total serum protein is not had obvious influence.
The tectology observed result
1) normal control group
Perusal: the kidney kermesinus, smooth surface, the kidney volume is not seen increase, the tangent plane skin, the medullary substance boundary is clear, does not see the expansion of calculus and renal pelvis.
Om observation: tunicle is smooth; Skin, medullary substance are high-visible, and renal corpuscle do not see and reduce or fibrosis, and far away, proximal convoluted tubule and collecting tubule epithelial cell are intact; Tube chamber is not seen calculus, cast and expansion; Matter does not see that cell infiltration and proliferation of fibrous tissue, renal pelvis divide a word with a hyphen at the end of a line between kidney, and epithelium is complete, and renal pelvis does not see that expansion, hydrops and JISHI form.
2) model control group:
Perusal: pale, the enlargement of kidney, smooth surface is graininess, and tangent plane is also pale, sees many large and small micropores that do not wait, and skin, medullary substance boundary are unclear, are dispersed in the tiny calculus of volume, the pyelectasis of minority kidney (2/8).
Light microscopy checking: tunicle is uneven, and the subregion thickens, and skin, medullary substance boundary are unclear; Matter was visible between glomerule, tubule reached is dispersed in irregularly shaped crystal aggregation in a large number, and the part renal tubules is blocked, and the part renal tubules is obviously expanded; Numbers of glomeruli obviously reduces, and the part atrophy, the nephron of no abnormality seen; Only island exists, and the part renal tubules is seen cast, and the part renal cells changes; Even necrosis comes off in the chamber matter visible foreign matters property granuloma and fibrosis between kidney, the renal pelvis expansion of part kidney.
3) sulfated fucan 200mg/kg group (1/8), the expansion of low-molecular-weight algal polysaccharide sulfate 100mg/kg group (0/8) renal pelvis, nephropathy only shows slightly lightly roughly like model control group.
4) low-molecular-weight algal polysaccharide sulfate 200mg/kg group:
Nephropathy and model control group are apparent in view to be alleviated, and most kidneys (5/9) skin, medullary substance boundary be not identification as yet, and numbers of glomeruli only slightly reduces, and remaining normal nephron zone is more obvious, does not see the renal pelvis expansion.
5) positive drug control group: dexamethasone acetate 0.07mg/kg.
Nephropathy is roughly organized like low-molecular-weight algal polysaccharide sulfate 200mg/kg, also compares with model control group obviously to alleviate.
The rat chronic renal failure pathological examination that above adenine causes shows: low-molecular-weight algal polysaccharide sulfate 200mg/kg and positive drug control group dexamethasone acetate all show the pathology that improve renal damage to be changed.
Experiment conclusion
1. low-molecular-weight algal polysaccharide sulfate 200,100mg/kg treat administration to rat; Irritate stomach every day once; Continuous 30 days; With model control group relatively, 200, the 100mg/kg dose groups obviously reduces kidney and partly excise induced CRF rat blood serum creatinine, urea nitrogen levels and glomerule area, improve renal tubules pathology and change degree.And with the dosage increase, effect strengthens.Show low-molecular-weight algal polysaccharide sulfate have the treatment kidney partly excise due to the effect of rat chronic renal failure.
2. low-molecular-weight algal polysaccharide sulfate 200,100mg/kg treat administration to rat, irritate stomach every day once, continuous 40 days; With model control group relatively, 200, the 100mg/kg dose groups obviously reduces freezing induced CRF rat blood serum creatinine, urea nitrogen levels; The 200mg/kg dose groups obviously reduces the glomerule area; Improve renal tubules pathology and change degree, and increase with dosage, effect strengthens.Show low-molecular-weight algal polysaccharide sulfate have treatment freezing due to the effect of rat chronic renal failure.
3. low-molecular-weight algal polysaccharide sulfate 200,100mg/kg irritate stomach every day once, continuous 40 days to rat prevention and treatment administration; Compare with model control group; 200, the 100mg/kg dose groups obviously reduces adenine induced CRF rat blood serum creatinine, urea nitrogen levels, and the 200mg/kg dose groups obviously increases albumin level, alleviates the kidney pathological change; And with the metering increase, effect strengthens.Show that low-molecular-weight algal polysaccharide sulfate has preventive and therapeutic effect to rat chronic renal failure due to the adenine.
More than the used positive control drug dexamethasone acetate 0.07mg/kg of experiment also has similar effect, and is suitable with 200mg/kg low-molecular-weight algal polysaccharide sulfate action intensity.
Embodiment 5 low-molecular-weight algal polysaccharide sulfates (sample A) are to the brightic observation of curative effect of rat Heymann
Choose Wistar kind rat, open the abdominal cavity after the stunning, insert syringe needle from renal artery, washing repeatedly to the kidney color with normal saline changes thick in vain; Take kidney, get renal cortex 5 grams and be ground into homogenate, be mixed into 10ml, add normal saline 20ml with Freund's complete adjuvant; Do intraperitoneal injection for rat of the same race, two weeks 1 time, each 2ml, about 3-6 time; Until albuminuria occurring, explain that model forms, and is divided into model control group with this rat; Positive drug control group is dexamethasone acetate 0.1mg/kg, low-molecular-weight algal polysaccharide sulfate A200, two dose groups of 100mg/kg and sample 1 (200mg/kg) group, and other establishes normal control group (not modeling).Gastric infusion, every day 1 time, model control group and normal control group wait the capacity distilled water; Altogether; Deliberated index: 1. urine protein quantitation: moulding before measurement 1 time, and remove the high person of albumen, confirm that surveying 1-3 time weekly after nephritis forms (puts into metabolic cage and get urine; Measure the twenty-four-hour urine protein quantification) 2. serum creatinine and blood urea nitrogen: before the moulding, confirm that nephritis forms that each was surveyed once before back and experiment finished.Adopt Beckman Model42 automatic biochemistry analyzer to measure.3. pathological examination.When experiment finishes, get kidney and carry out kidney segment (HE dyeing), light microscopy checking, every example is selected the glomerule of 5 maximum tangent planes, metering glomerule diameter and cell number.
Experimental result: 4 weeks of low-molecular-weight algal polysaccharide sulfate gastric infusion are listed table 6 in to the influence of rat Heymann nephritis urine protein, and the influence of serum creatinine, blood urea nitrogen is listed in table 7, and the pathological examination result lists table 8 in.
Table 6 low-molecular-weight algal polysaccharide sulfate is to the influence of rat Heymann nephritis urine protein
N: number of animals; △ p<0.05, △ △ p<0.01, △ △ △ p<(0.001 with comparing before the moulding);
*p<0.05,
*p<(0.01 comparing) with model control group.
Visible by table 6; Rat urine protein quantitation behind each assembly molding, all (P < 0.001) shows the model success before moulding, and urine protein raises and continues more than 4 weeks; Low-molecular-weight algal polysaccharide sulfate 200mg/>kg and 100mg/kg are in treatment 4 weeks of back; Make rat urine protein eliminating amount obviously reduce (with model control group relatively P value respectively 0.05 and 0.01), the positive control drug dexamethasone acetate also makes the rat urine protein that certain therapeutical effect is arranged, and strengthens with dosage increase effect.
Table 7 low-molecular-weight algal polysaccharide sulfate is to the influence of rat Heymann nephritis serum creatinine, blood urea nitrogen
N: number of animals; △ p<0.05, △ △ p<0.01, △ △ △ p<(0.001 with comparing before the moulding);
*p<(0.01 comparing) with model control group.
Table 7 is the result show; After the moulding rat blood serum creatinine and blood urea nitrogen obviously raise before than moulding (p 0.01 or p 0.001); Low-molecular-weight algal polysaccharide sulfate (200 and 100mg/kg) and positive drug control group treatment back serum creatinine obviously descend (with model control group relatively p 0.01), serum urea nitrogen is not had obvious influence.Experimental result shows that low-molecular-weight algal polysaccharide sulfate has certain therapeutical effect to rat Heymann nephritis serum creatinine, increases with dosage, and effect strengthens.
Table 8 low-molecular-weight algal polysaccharide sulfate is to the pathological influence of rat Heymann nephritis kidney
| Group | Dosage (mg/kg) | Number of animals (only) | Glomerule diameter (lattice number) | Messangial cell number (number) |
| Blank | 10 | 8.62±0.96 | 87.03±8.34 | |
| The model contrast | 8 | △△△11.12±0.83 | △△△116.10±15.37 | |
| Sample A | 200 | 8 | 9.71±1.05 ** | 109.99±38.97 |
| Sample A | 100 | 8 | 10.75±0.88 | 119.24±14.39 |
| Sample 1 | 200 | 8 | 10.35±1.09 | 117.30±16.80 |
| Dexamethasone acetate | 0.1 | 8 | 9.76±0.68 ** | 105.79±9.27 |
△ △ △ p<(0.001 with comparing before the moulding);
*p<(0.01 comparing) with model control group
Table 8 result shows that rat pathological examination glomerule diameter and messangial cell number average are apparently higher than the blank group after the moulding.Low-molecular-weight algal polysaccharide sulfate administration group has reduction trend to the glomerule diameter, and messangial cell is not had obvious influence.The positive control drug dexamethasone acetate can obviously reduce the glomerule diameter.
Experiment conclusion
Low-molecular-weight algal polysaccharide sulfate 200,100mg/kg in 4 weeks of gastric infusion, can reduce rat Heymann nephritis model urine protein output and serum creatinine value; And increase with dosage; Effect strengthens, and serum urea nitrogen is not had obvious influence, pathological examination; The glomerule diameter there is reduction trend, the messangial cell number is not had obvious influence.
Embodiment 6 low-molecular-weight algal polysaccharide sulfates (sample C) are to diuresis normal and water load rat
1, to the effect of water load animal
The male and healthy Wistar rat of body weight 150g～200g placed metabolic cage pre-adaptation 1, accessed 6 hours urines, and whether the urine amount of observing under the free conditions of water drinking is stable.Press 2.5ml/100g again and irritate the stomach normal saline, the urine amount of collecting in 2 hours reaches the selected test of animal of the amount of pouring into 40%.
By 6 hours voided volume animal is divided into 6 groups at random, i.e. low, the high dose group of low-molecular-weight algal polysaccharide sulfate (sample C), sample 1 (200mg/kg) group, hydrochlorothiazide group, model group, normal group.Administration group dosage is respectively low-molecular-weight algal polysaccharide sulfate C 100,200mg/kg, hydrochlorothiazide 4mg/kg, and model group gives the isometric(al) normal saline, and normal group does not process.
The animal fasting is 18 hours before the test.Administration treated animal medicine mixes in load water, (to irritate stomach by 2.5ml/100g and give 38 ℃ of normal saline) and gives during test.Model group is with isometric physiologic saline for substitute medicinal liquid.Animal is put into rapidly and gives the fully metabolic cage of drinking-water after the administration, places 1 of rat in every metabolic cage, gently presses the thing lower abdomen before putting into, and drains surplus urine.Per hour connect urine once behind the medicine, observed continuously 6 hours, observe the difference of respectively organizing urine amount in the certain hour.
The result representes with mean ± standard deviation, with t check carrying out test of significance between group.See table 9
Table 9 low-molecular-weight algal polysaccharide sulfate is to water load rat unit urine amount in the time (unit: (x ± s) of influence ml)
.vs normal group .p<0.05 ..p<0.01
*The vs model group
*p<0.05
*p<0.01
Visible by table 9, model group gives water load back animal urine amount obviously to be increased, and with normal group significant difference (p < 0.01 or 0.05) is arranged relatively during 0～3 hour.
Each group of low-molecular-weight algal polysaccharide sulfate after administration in 6 hours urine amount and model group increase is more all arranged, wherein 100mg/kg organizes during after the administration 3～4 hours the urine amount has significant difference (p < 0.05); 200mg/>kg group urine amount during after the administration 1～6 hour has significant difference (p < 0.01 or 0.05).Sulfated fucan 200mg/>kg group also has effect trend.Each dose groups effect has dose dependent.
Each group of low-molecular-weight algal polysaccharide sulfate 2～4 hours diuresis after administration are the strongest.
The hydrochlorothiazide group after administration in 6 hours urine amount and model group significant difference (p < 0.01 or 0.05) is more all arranged.
2. to intact animal's effect
The male and healthy Wistar rat of body weight 150～200g placed metabolic cage pre-adaptation 1, accessed 6 hours urines, observed the urine amount under the free conditions of water drinking, selected the selected test of the stable animal of urine amount.
By 6 hours voided volume animal is divided into 5 groups at random, promptly receives reagent object height, low dose group, 1 group in sample, hydrochlorothiazide group, normal group.Administration group dosage is respectively low-molecular-weight algal polysaccharide sulfate 100,200mg/kg, sulfated fucan (sample 1) 200mg/kg, hydrochlorothiazide 4mg/kg.The administration volume is 1ml/100g, and normal group gives the isometric(al) solvent.
The animal fasting is 18 hours before the test.Animal is put into the metabolic cage that gives sufficient drinking-water rapidly after the administration, places 1 of rat in every metabolic cage, gently presses the thing lower abdomen before putting into, and drains surplus urine.Per hour connect urine once behind the medicine, observed continuously 6 hours, observe the difference of respectively organizing urine amount in the certain hour.The result representes with mean ± standard deviation, carries out test of significance with t check between group, sees table 10.
Table 10 low-molecular-weight algal polysaccharide sulfate is to normal rat unit urine amount in the time (unit: (x ± s) of influence ml)
*The vs normal group
*p<0.05
*p<0.01
With normal group increase is arranged more all by each group of the visible low-molecular-weight algal polysaccharide sulfate of table 10 urine amount after administration, wherein 200mg/kg organizes during after the administration 0～5 hour urine amount and normal group relatively has significant difference (p < 0.01 or 0.05); 100mg/>kg group urine amount in the time of administration rear section also has significant difference (p < 0.01 or 0.05).Each dose groups effect has dose dependent.
Each group of low-molecular-weight algal polysaccharide sulfate 2～4 hours diuresis after administration are the strongest.
The hydrochlorothiazide group after administration in 6 hours urine amount and normal group significant difference (p < 0.01 or 0.05) is more all arranged.
Conclusion (of pressure testing): low-molecular-weight algal polysaccharide sulfate 100～200mg/kg single gastric infusion all has diuresis to normal and water load rat; 2～4 hours diuresis are the strongest after administration; Each dose groups difference between the effects is obvious, has the doses dependence.
Embodiment 7 low-molecular-weight algal polysaccharide sulfates (sample B) are to the influence of mice delayed hypersensitivity and humoral immunization
The healthy kunming mice of body weight 24～26g, the male and female dual-purpose.With 1.25% 2, the 4-dinitrochlorobenzene acetone solution subcutaneous 0.02ml/ in back only injects mouse sensitization.Sensitization one day after, animal is divided into 5 groups at random, promptly two dose groups of low-molecular-weight algal polysaccharide sulfate B and sample are 1 group, dexamethasone acetate group, model group.Irritate stomach respectively and give low-molecular-weight algal polysaccharide sulfate 150,300mg/kg, sulfated fucan 300mg/kg, dexamethasone acetate 0.1mg/kg; Model group gives the isometric(al) normal saline, continuous irrigation stomach 9 days, and the 10th day with 0.25%2; In the middle of the right foot pad of 4-dinitrochlorobenzene acetone solution subcutaneous injection mice, dosage is 0.02ml/, and parapodum injection isometric(al) acetone solution is compared; Cut two foots from ankle joint after 38 hours; Scales/electronic balance weighing according to computes swelling degree and suppression ratio, and carries out statistical procedures.
Swelling degree=right heavy sensation in the foot amount (mg)-left heavy sensation in the foot amount (mg)
It is centrifugal to get angular vein blood before the execution animal, rate scattering turbidimetry for Determination change of serum C 3, C4, IgG, IgM content.
Other gets 10 Kunming mouses as normal group, raise 10 days equally after, it is centrifugal that angular vein is got blood, the same survey change of serum C 3, C4, IgG, IgM content.The result sees table 11,12.
Table 11 low-molecular-weight algal polysaccharide sulfate is to 2, the influence of 4-dinitrochlorobenzene induced mice delayed hypersensitivity
| Group | Dosage (mg/kg) | Number of animals (only) | The swelling degree (
 |
Suppression ratio (%) | P value (vs model group) |
| Model | \ | 10 | 45.58±28.24 | ||
| Sample 1 | 300 | 8 | 30.92±22.25 | 36.36 | >0.05 |
| Sample B | 150 | 9 | 26.30±12.08 | 45.86 | <0.01 |
| Sample B | 300 | 9 | 23.10±4.43 | 52.45 | <0.01 |
| Dexamethasone | 0.1 | 9 | 15.96±11.20 | 67.15 | <0.01 |
Visible by table 11,4-dinitrochlorobenzene induced mice delayed hypersensitivity had inhibitory action to 2 in 9 days in low-molecular-weight algal polysaccharide sulfate ig administration, and 150,300mg/kg group swelling degree and model group comparison p < 0.01.The dexamethasone acetate group also has similar effect.
Table 12 low-molecular-weight algal polysaccharide sulfate is to change of serum C
3, immunoglobulin, total complement content influence
| Group | Dosage (mg/kg) | Number of animals (only) | C 3 | C 3 | 1gG | 1gM |
| Normally | \ | 8 | 35.63±4.94 | 24.59±10.27 | 27.33±12.92 | 81.78±40.44 |
| Model | \ | 8 | 51.79±26.71 | 30.92±7.07· | 36.89±8.60·· | 103.33±20.26· |
| Sample 1 | 300 | 8 | 54.96±29.49· | 28.58±5.51 | 37.43±10.14 | 92.75±25.58 |
| Sample B | 150 | 7 | 37.23±24.92 | 23.43±11.10 * | 37.96±6.19 | 112.12±29.74 |
| Sample B | 300 | 8 | 35.01±18.04 * | 16.02±10.56 ** | 30.21±15.43 * | 80.23±28.15 * |
| Dexamethasone | 0.1 | 9 | 30.03±7.85 ** | 17.95±9.82 ** | 31.08±7.37 * | 79.32±26.80 * |
Vs normal group p<0.05p<0.01
*The vs model group
*p<0.05
*p<0.01
Visible by table 12, the humoral immunization index all raises behind mouse subcutaneous injection 2,4 dinitrochlorobenzene.Give medicine ig after 9 days, low-molecular-weight algal polysaccharide sulfate 150mg/kg group has inhibitory action to the mouse humoral immune part index number; Low-molecular-weight algal polysaccharide sulfate 300mg/kg group and dexamethasone acetate group all have remarkable inhibitory action to mouse humoral immune each item index, with model group p < 0.05 or p < 0.01 relatively.
Conclusion (of pressure testing): (1) low-molecular-weight algal polysaccharide sulfate 150,300mg/kg administration ig9 days have obvious inhibitory action to 2,4 dinitrochlorobenzene induced mice delayed hypersensitivitys, and dose dependent is arranged; (2) low-molecular-weight algal polysaccharide sulfate 150,300mg/kg administration can obviously suppress the allergy mouse humoral immune in ig9 days.
Claims (9)
1. the purposes of low-molecular-weight algal polysaccharide sulfate in the medicine of preparation treatment kidney disease; Wherein said low-molecular-weight algal polysaccharide sulfate is sulfated polysaccharides or the oligosaccharide class material that sulfated fucan is made through degraded, and wherein said low-molecular-weight algal polysaccharide sulfate molecular weight is 1000～200000.
2. according to the purposes of claim 1, wherein said kidney disease is selected from one or more in renal failure, nephritis and the nephrotic syndrome.
3. according to the purposes of claim 2, wherein said nephritis is one or more in common clinically acute glomerulonephritis, chronic glomerulonephritis, pyelonephritis, latent nephritis, Henoch Schonlein purpura nephritis, the lupus erythematosus nephritis.
4. according to the purposes of each claim among the claim 1-3, wherein said low-molecular-weight algal polysaccharide sulfate molecular weight is 5000～100000.
5. according to the purposes of claim 4, wherein said low-molecular-weight algal polysaccharide sulfate molecular weight is 8000～80000.
6. according to the purposes of claim 5, wherein said low-molecular-weight algal polysaccharide sulfate molecular weight is 50000～80000.
7. according to the purposes of claim 5, wherein said low-molecular-weight algal polysaccharide sulfate molecular weight is 15000～30000.
8. according to the purposes of claim 1, sulfated fucan wherein derive from Thallus Laminariae (Thallus Eckloniae), wild Brown algae Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, Mus tail algae, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae), yellow tang or Fucus Vesiculosus one or more.
9. according to the purposes of claim 1, the dosage form of low-molecular-weight algal polysaccharide sulfate wherein is injection, oral formulations, local administration preparation or nasal cavity administrated prepn.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007100643133A CN101028282B (en) | 2007-03-12 | 2007-03-12 | Use of low-molecular weight phaeophytin polyose sulfate in preparation of medicine for treating kidney disease |
| PCT/CN2008/000850 WO2008122214A1 (en) | 2007-03-12 | 2008-04-25 | The use of low molecular weight fucoidan in preparing of medicine for treating kidney diseases |
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| CN2007100643133A CN101028282B (en) | 2007-03-12 | 2007-03-12 | Use of low-molecular weight phaeophytin polyose sulfate in preparation of medicine for treating kidney disease |
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| CN101306014B (en) * | 2008-04-03 | 2012-03-14 | 北京世纪博康医药科技有限公司 | Use of brown algae polysaccharide sulfuric acid ester of low molecular weight in preparing medicine for treating diabetic nephropathy |
| CN102532332B (en) * | 2011-09-09 | 2014-07-09 | 山东洁晶集团股份有限公司 | Method for extracting and preparing low molecular weight fucoidin from marine brown algae |
| CN107899005B (en) * | 2017-11-20 | 2021-02-26 | 北京颐方生物科技有限公司 | Composition containing fucoidan, tablet and application thereof |
| CN111494407A (en) * | 2020-01-08 | 2020-08-07 | 南京市儿童医院 | Use of trehalose in the preparation of a medicament for alleviating conditions associated with acute renal injury induced by ischemia reperfusion |
| CN115671118B (en) * | 2021-07-22 | 2024-02-27 | 海糖（江苏）生物医药科技有限公司 | Application of brown alginate oligosaccharides |
| CN113632987A (en) * | 2021-08-12 | 2021-11-12 | 威海威仕泰医疗科技有限公司 | Kidney disease non-total nutrient special medical food based on fucoidin and preparation method thereof |
| CN114984037B (en) * | 2022-08-04 | 2022-10-04 | 山东第一医科大学附属省立医院(山东省立医院) | Low-molecular-weight algin and preparation method and application thereof |
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| CN1129109A (en) * | 1995-12-16 | 1996-08-21 | 中国科学院海洋研究所 | Medicine of alginic polysaccharide sulfate for treatment of nephrasthenia and angiopathy and preparation method thereof |
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| CN1206591A (en) * | 1998-05-08 | 1999-02-03 | 毕爱芳 | Medicine for brown algae polyose sulfate for curing glomerular disease |
| CN1069523C (en) * | 1998-10-14 | 2001-08-15 | 中国科学院海洋研究所 | Application of alginic polysaccharide sulfate in preparation of medicine for treating renal failure |
| CN1267457C (en) * | 2004-10-18 | 2006-08-02 | 中国科学院海洋研究所 | Method for preparing low molecular weight seaweed polysaccharide sulfate |
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| CN1129109A (en) * | 1995-12-16 | 1996-08-21 | 中国科学院海洋研究所 | Medicine of alginic polysaccharide sulfate for treatment of nephrasthenia and angiopathy and preparation method thereof |
Non-Patent Citations (2)
| Title |
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| 吴茜茜等.功能性低分子量岩藻多糖的研究进展.生物学杂志23 3.2006,23(3),4-7. |
| 吴茜茜等.功能性低分子量岩藻多糖的研究进展.生物学杂志23 3.2006,23(3),4-7. * |
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