CN101530458B - Application and preparing method of Chinese milkvetch seed extractive - Google Patents
Application and preparing method of Chinese milkvetch seed extractive Download PDFInfo
- Publication number
- CN101530458B CN101530458B CN2009100977218A CN200910097721A CN101530458B CN 101530458 B CN101530458 B CN 101530458B CN 2009100977218 A CN2009100977218 A CN 2009100977218A CN 200910097721 A CN200910097721 A CN 200910097721A CN 101530458 B CN101530458 B CN 101530458B
- Authority
- CN
- China
- Prior art keywords
- extract
- ethanol
- milkvetch seed
- amount
- concentrated solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Abstract
The invention discloses an application and preparing method of Chinese milkvetch seed extractive. Triterpenes, flavonoids and their glycosides as effective components are extracted out from Chinese milkvetch seed as raw material and used for reducing the urokinase protein and treating the nephrotic syndrome. Chinese milkvetch seed is extracted by the ethanol reflux extraction method, the solvent is recycled under decompression, the triterpenes, flavonoids and their glycosides are obtained after the macroporous polymeric adsorbent column chromatography or the n-butanol extraction separation. The Chinese milkvetch seed extractive has the functions of reducing the urokinase protein and treating the nephrotic syndrome, and can be taken for a long time without obvious noxious and side effect; the process is reasonable and simple, the effective components are remained, the dosage is reduced.
Description
Technical field
The present invention relates to the natural drug technology, particularly relate to a kind of purposes and preparation method of Chinese milkvetch seed extractive.
Background technology
Kidney is the device of wanting of the five internal organs, the congenital foundation, its effect is to discharge the intravital metabolite of people, the useful component in the metabolite is reuptaked in the ex vivo, again produce refuse---urine is responsible for discharge by ureter, bladder.Simultaneously, kidney still is very important endocrine organ in the body, is responsible for making the erythropoietin that promotes hemopoietic, and activity of vitamin d3 also is responsible for the decomposition of insulin in the body.In case renal function suffers damage, refuse is accumulated in vivo, then can produce very big harm to health.
At present China kidney patient has reached 9,000 ten thousand people, and wherein, uremic patient every year is just with the speed increment of 5%-10%.According to data, since 2006, chronic nephropathy has become the fourth-largest disease killer after cardiovascular and cerebrovascular disease, tumor, diabetes, and from global general status, the control of chronic kidney disease is faced with stern challenge.And in chronic renal disease, nephrotic syndrome has accounted for very big proportion
[1]
(nephrotic syndrome NS) is the common disease of Urology Department, multiple disease to nephrotic syndrome! A lot of kidney diseases all show as nephrotic syndrome clinically
[2], it is not an independence disease, but by multiple reason, comprises one group of clinical syndrome in the renal glomerular disease that chronic nephritis causes.The typical case shows as a large amount of albuminuria, hypoalbuminemia, edema, hyperlipemia.Because hypoproteinemia, hyperlipemia and edema all are a large amount of albuminuretic consequences, therefore, the standard of diagnosis should be based on a large amount of albuminuria
[3]And a large amount of albuminuria are the important independently risk factors that cause renal function to worsen, no matter be constitutional or Secondary cases nephrotic syndrome, if a large amount of albuminuria can not get controlling timely and effectively, to cause nephridial tissue, particularly between kidney matter carry out fibrosis, carrying out property of renal function goes down, and develops into renal failure in latter stage at end gradually
[1]Therefore, its treatment key is to reduce or eliminate albuminuria
[2]
Aspect its treatment measure, adrenocortical hormone is the choice drug of treatment nephrotic syndrome, and as prednisone, prednisolone, dexamethasone etc., but it is a lot of to take such drug side effect for a long time, quite serious sometimes
[3]Simultaneously, the part patient exists hormone dependence, hormonal resistance, hormone not to tolerate and phenomenon such as outbreak repeatedly
[1]This a part of patient can try out cytotoxic drug, be situated between as cyclophosphamide and benzenebutanoic acid nitrogen, but the same toxic and side effects of this type of medicine is very big and how the recidivist does not advocate to carry out the medication second time to alleviating again after the medication, in order to avoid poison
[3]In addition, also can select cellular immunization inhibitor---Ciclosporin A for use, but this medicine also there is serious toxic and side effects, the most serious is liver, nephrotoxicity, its nephrotoxicity incidence rate is at 20%-40%, thus should not be for a long time with this medicine treatment nephrotic syndrome, more should not be easily with this medicine as choice drug
[3]Because of the limitation of above-mentioned various kinds of drug existence and serious toxic and side effects, used by people gradually in conjunction with the traditional Chinese medical herbal treatment nephrotic syndrome, but up till now, still there is not a species specific efficient medicine of low toxicity that is widely used in nephrotic syndrome and reduces urine protein.
Herba Astragali Melilotoidis (Herba Astragali Sinici) (Astragalus sinicus Linn.) is a pulse family Radix Astragali platymiscium, is distributed in each provinces and regions, the Yangtze river basin, is longer than the hillside sylvan life, limes marginis and meadow.Medicinal part is herb and seed, the function that has heat-clearing and toxic substances removing, makes eye bright and invigorate blood circulation
[4]Semen astragali sinici kidney shape or rhombus contain the flavone and the glycoside material thereof of Wheat Protein according to the literature
[5], triterpene glycoside material
[6], materials such as aspartic acid and polysaccharide.But up to now, do not see its modern pharmacological research report.
Above-mentioned unfavorable present situation in view of present nephrotic syndrome clinical application, the present invention utilizes the mRNA IN ADRIAMYCIN NEPHROPATHY rat model that multiple natural plant extracts has been carried out pharmacological experiment study, observe of the influence of these extracts to urine protein content, it is unexpected that the discovery Chinese milkvetch seed extractive have remarkable reduction urine protein, improve the pharmacological action of the nephrotic syndrome, effect and prednisone are suitable, and have no side effect.Content of the present invention is not seen relevant report both at home and abroad.
[1] Yuan Fahuan.Intractable nephrotic syndrome [J].Chongqing medical science, 2008,37 (8): 786-788.
[2] Jin Ping.The progress of nephrotic syndrome medicine [J].Foreign medical science urinary system fascicle, 2001,21 (4): 145-147.
[3] nephrotic syndrome.Nephropathy medical science net, http://www.shenbing.org.
[4] " Chinese Plants will ", the chief editor of Chinese Plants will editorial board of the Chinese Academy of Sciences, Science Press publishes, and 42 (1): 199.
[5]Yeom，SH；Kim，MK；Kim，HJ；Shim，JG；Lee，JH；Lee，MW.Phenoliccompounds?from?seeds?of?Astragalus?sinicus?and?their?antioxidativeactivities[J].Saengyak?Hakhoechi，2003，34(4)：344-351.
[6]Cui，B；Inoue，J；Takeshita，T；Kinjo，J；Nohara，T；Takeshita，T；Kinjo，J；Nohara，T.Triterpene?glycosides?from?the?seeds?of?Astragalussinicus?L[J].Chemical?&?Pharmaceutical?bul?let?in，1992，40(12)：3330-3.
Summary of the invention
The object of the present invention is to provide a kind of reduction urine protein, natural drug extract of treatment nephrotic syndrome and preparation method thereof.
The technical solution used in the present invention is as follows:
One, a kind of purposes of Chinese milkvetch seed extractive:
This extract is an effective ingredient with triterpene, flavone and their glycoside material, is used to reduce urine protein, the treatment nephrotic syndrome.
Two, a kind of preparation method of Chinese milkvetch seed extractive, the step of this method is as follows:
1) Semen astragali sinici parching to brown is pulverized, and crosses 10 mesh sieves, adds 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, and each 2h filters, merging filtrate, decompression recycling ethanol must extract concentrated solution;
2) should extract concentrated solution adsorbs with macroporous adsorptive resins, after washing post with 6 times of water gagings and 7 times of amount 30% ethanol successively, with 9 times of amount 70% ethanol elutions, the eluent decompression and solvent recovery, at 60 ℃ of following vacuum dryings, get dry extract, dry extract is broken into fine powder, just be prepared into extract of the present invention.
Described step 2) big pore adsorption resin comprises nonpolar, low pole and middle polarity macroporous adsorbent resin in.
Three, the preparation method of another kind of Chinese milkvetch seed extractive, the step of this method is as follows:
1) Semen astragali sinici parching to brown is pulverized, and crosses 10 mesh sieves, adds 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, and each 2h filters, merging filtrate, decompression recycling ethanol must extract concentrated solution;
2) should extract concentrated solution, successively with petroleum ether, the extraction of ethyl acetate equal-volume, after each extracts 3 times,, merge butanol extraction liquid with equal-volume n-butanol extraction 3 times, decompression and solvent recovery, at 60 ℃ of following vacuum dryings, in drying baker, place the residual n-butyl alcohol of activated carbon adsorption, get dry extract, dry extract is broken into fine powder, just has been prepared into extract of the present invention.
N-butyl alcohol extract and macroporous adsorbent resin 70% ethanol elution thing carry out chromogenic reaction:
(1) hydrochloric acid-magnesium powder reaction: the positive shows and contains flavone compound.
(2) Molish reaction: the positive shows and contains glycosides compound.
(3) L-B reaction: the positive shows and contains triterpenoid compound.
Above-mentioned chromogenic reaction in conjunction with document as can be known this Chinese milkvetch seed extractive be effective ingredient with flavone, triterpene and their glycoside material.
The beneficial effect that the present invention has is:
1. Chinese milkvetch seed extractive has remarkable reduction urine protein, the effect of treatment nephrotic syndrome;
2. Chinese milkvetch seed extractive does not have obvious toxic-side effects.Treat nephrotic syndrome at present clinically, reduce the urine egg and use hormone medicine and immunosuppressive drug more, toxic and side effects is big, and Chinese milkvetch seed extractive does not have obvious toxic-side effects, can take for a long time.
3. technology is rationally easy, can keep effective ingredient to greatest extent, reduces dosage.
The specific embodiment
Be embodiments of the invention below, but the present invention is not limited thereto.
Embodiment 1:
Semen astragali sinici 2.5kg, parching to brown is pulverized, cross 10 mesh sieves, add 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, each 2h filters merging filtrate with 2 layers of hospital gauze, decompression (0.08-0.10MPa) extracts respectively 3 times with equal-volume petroleum ether, ethyl acetate and n-butyl alcohol behind the recovery ethanol successively, decompression (0.08-0.10MPa) is reclaimed solvent, at 60 ℃ of following vacuum dryings, is placed the residual n-butyl alcohol of activated carbon adsorption in drying baker, get dry extract, dry extract is broken into fine powder preserves.
Embodiment 2
Semen astragali sinici 28kg, parching to brown, pulverize, cross 10 mesh sieves, add 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, each 2h, filter with 2 layers of hospital gauze, merging filtrate, decompression (0.08-0.10MPa) reclaim behind the ethanol with the D101 absorption with macroporous adsorbent resin, wash post with 6 times of water gagings and 7 times of amount 30% ethanol after, with 9 times of amount 70% ethanol elutions, wash post with 95% ethanol at last, the eluent decompression (0.08-0.10MPa) is reclaimed solvent, at 60 ℃ of following vacuum dryings, get dry extract, dry extract is broken into fine powder preserves.
Correlational study result of the present invention is as follows:
One, among the embodiment 1 each extracted component to the influence of mRNA IN ADRIAMYCIN NEPHROPATHY rat model
1, laboratory animal: the healthy SD rat, male, body weight 350 ± 20g is available from Zhejiang Province's Experimental Animal Center.The quality certification number: SCXK (Zhejiang 2003-0001).
2, experiment given the test agent and medicine:
1) each extracted component of Semen astragali sinici: each extracted component is that the ultrasonic suspendible of medium is irritated stomach with water among the embodiment 1.
2) prednisone: Zhejiang Province XianJu Pharmacy stock Co., Ltd produces, lot number 0704148, specification 5mg.
3) doxorubicin hydrochloride: the production of Mingzhi's medicine company limited, lot number 070101, specification 10mg.
3, animal grouping and dosage:
Rat is divided into 7 groups at random, 10 every group, is respectively each the extraction battery grouping of blank group, model group, positive controls (prednisone) and Semen astragali sinici; The prednisone dosage is 15mg/kg, and each group of Semen astragali sinici is grouped into and is equivalent to 10g raw medicinal herbs/kg, and it is 10ml/kg that model group gives isopyknic distilled water.
4, experimental technique:
Blank group rat tail vein injecting normal saline, other six groups of tail vein injection amycin 5mg/kg modelings began gastric infusion in modeling the same day, and once a day, continuous 28 days, blank group and model group were irritated stomach and are given the equivalent distilled water.Administration 28d claims each Mus weight, and rat is positioned over collection 24h urinary assay urine protein content in the metabolic cage.
5, urine protein content assay method: Coomassie brilliant blue method
1) preparation of bovine serum albumin standard solution: accurately take by weighing the 100mg bovine serum albumin, be dissolved in the 100mL distilled water, be the stock solution of 1000 μ g/mL.
2) preparation of protein reagent Coomassie brilliant blue G-250: take by weighing 100mg Coomassie brilliant blue G-250, be dissolved in 50mL 90% ethanol, add the phosphatase 11 00mL of 85% (W/V), use the distilled water standardize solution at last to 1000mL.
3) sample determination: urine to be measured centrifugal 20 minutes in 4000r/min, get supernatant 1ml and put in the tool plug test tube, add the 5ml protein reagent, fully mix, placed 10 minutes, and, checked in Protein content in the testing sample by standard curve with 1cm optical path cuvette colorimetric record under 595nm.
6, statistical procedures: experimental data adopts SPSS 12.0 softwares to handle with average (X) expression, whole experimental datas.
7, experimental result: each group rat urine protein content compared to find that n-butyl alcohol extract has the effect of the reduction mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine protein similar to prednisone, compare with model group all to have statistical significance.And all the other each components there is no any influence to the rat urine protein content.Concrete experimental result sees Table 1.
Table 1 Semen astragali sinici respectively extracts the influence (n=10) of component to mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine albumen amount
Annotate: P is the probability of comparing with model group
Two, among the embodiment 2 each component to the influence of mRNA IN ADRIAMYCIN NEPHROPATHY rat model
1, laboratory animal: the healthy SD rat, male, body weight 350 ± 20g is available from Zhejiang Province's Experimental Animal Center.The quality certification number: SCXK (Zhejiang 2003-0001).
2, experiment given the test agent and medicine:
1) each component of Semen astragali sinici: macroporous adsorptive resins washing post partly is designated as ASA among the embodiment 2, and 30% washes post partly is designated as ASB, and 70% eluting partly is designated as ASC, and 95% washes post partly is designated as ASD, and each several part is irritated stomach with the ultrasonic suspendible of water.
2) prednisone: Zhejiang Province XianJu Pharmacy stock Co., Ltd produces, lot number 0704148, specification 5mg.
3) doxorubicin hydrochloride: the production of Mingzhi's medicine company limited, lot number 070101, specification 10mg.
3, animal grouping and dosage:
Rat is divided into 7 groups at random, 10 every group, is respectively blank group, model group, positive controls (prednisone) and each component group of Semen astragali sinici; The prednisone dosage is 15mg/kg, and each group of Semen astragali sinici is grouped into and is equivalent to 10g raw medicinal herbs/kg, and it is 10ml/kg that model group gives isopyknic distilled water.
4, experimental technique:
Blank group rat tail vein injecting normal saline, other six groups of tail vein injection amycin 5mg/kg modelings began gastric infusion in modeling the same day, and once a day, continuous 28 days, blank group and model group were irritated stomach and are given the equivalent distilled water.Administration 28d claims each Mus weight, and rat is positioned over collection 24h urinary assay urine protein content (Coomassie brilliant blue method) in the metabolic cage.
5, statistical procedures: experimental data adopts SPSS 12.0 softwares to handle with average (X) expression, whole experimental datas.
6, experimental result: each group rat urine protein content compared to find that ASC is the effect that macroporous adsorbent resin 70% eluate has the reduction mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine protein similar to prednisone, compare with model group all to have statistical significance.And all the other each components there is no any influence to the rat urine protein content.Concrete experimental result sees Table 2.
Table 2 Semen astragali sinici each component is to the influence (n=10) of mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine albumen amount
Annotate: P is the probability of comparing with model group
Three, ASC is to the influence of mRNA IN ADRIAMYCIN NEPHROPATHY rat model
1, laboratory animal: the healthy SD rat, male, body weight 350 ± 20g is available from Zhejiang Province's Experimental Animal Center.The quality certification number: SCXK (Zhejiang 2003-0001).
2, experimental drug:
1) Chinese milkvetch seed extractive: ASC.
2) prednisone: Zhejiang Province XianJu Pharmacy stock Co., Ltd produces, lot number 0704148, specification 5mg.
3) doxorubicin hydrochloride: the production of Mingzhi's medicine company limited, lot number 070101, specification 10mg.
3, animal grouping and dosage:
Rat is divided into 4 groups at random, 10 every group, is respectively blank group, model group, positive controls (prednisone) and ASC group; The prednisone dosage is 15mg/kg, and ASC organizes to being equivalent to 10g raw medicinal herbs/kg, and it is 10ml/kg that model group gives isopyknic distilled water.
4, experimental technique:
Blank group rat tail vein injecting normal saline, other three groups of tail vein injection amycin 5mg/kg modelings began gastric infusion in modeling the same day, and once a day, continuous 28 days, blank group and model group were irritated stomach and are given the equivalent distilled water.Administration 28d claims each Mus weight, and rat is positioned over collection 24h urinary assay urine protein content (Coomassie brilliant blue method) in the metabolic cage.24h vena orbitalis posterior clump is got blood after the last administration, measures total protein, creatinine, blood urea nitrogen, T-CHOL and triglyceride in the blood in automatic biochemistry analyzer, gets kidney after disconnected neck is put to death and does histological examination.
5, renal tissue is learned inspection method: the tubulointerstitial injury degree is carried out semi-quantitative analysis (with reference to Journal American Society of Nephrology.1997,8 (2): method 199-207): to every part of specimen of each experimental group at low power lens 5 glomerule of blind observation that place an order, the matter visual field between renal tubules, be that observation is upper left under every specimen low power lens, upper right, the lower-left, the bottom right, renal tubules in middle 5 visuals field, matter between kidney, according to the renal cells vacuolar degeneration, tubular ectasia, interstitial fibrosis, between four indices such as matter cell infiltration to renal tubules, the kidney ID becomes marks, each index is divided into gently according to lesion degree, in, weigh three degree, remember 1 respectively, 2, calculated its average in 3 minutes.
6, statistical procedures: experimental data adopts SPSS 12.0 softwares to handle with average (X) expression, whole experimental datas.
7, experimental result: concrete experimental result sees Table 3-5.
Table 3ASC is to the influence (n=10) of mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine albumen amount
Annotate: P is the probability of comparing with model group
Relatively the data that go up in the table can determine further that Chinese milkvetch seed extractive has the effect of the reduction mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine protein similar to prednisone, compare with model group all to have statistical significance.
Table 4 blood physicochemical data result (n=10)
Annotate: * and model group compare, p＜0.05
Relatively go up data in the table and can find that Chinese milkvetch seed extractive can make in the blood of mRNA IN ADRIAMYCIN NEPHROPATHY rat model total protein content keep substantially normally, make creatinine Cr and content of triglyceride obtain to a certain degree reduction.
Table 5 histopathologic examination (n=10)
Annotate: * and model group compare, p＜0.05
Relatively the data that go up in the table can find that Chinese milkvetch seed extractive can make the pathological phenomenons such as cellular edema, tubular ectasia and interstitial fibrosis of mRNA IN ADRIAMYCIN NEPHROPATHY rat model be significantly improved.
Four, ASC is to excising the influence of mRNA IN ADRIAMYCIN NEPHROPATHY rat model behind the left kidney
1, laboratory animal: the healthy SD rat, male, body weight 350 ± 20g is available from Zhejiang Province's Experimental Animal Center.The quality certification number: SCXK (Zhejiang 2003-0001).
2, experiment is subjected to test product and medicine:
1) Chinese milkvetch seed extractive: ASC.
2) prednisone: Zhejiang Province XianJu Pharmacy stock Co., Ltd produces, lot number 0704148, specification 5mg.
3) doxorubicin hydrochloride: the production of Mingzhi's medicine company limited, lot number 070101, specification 10mg.
3, animal grouping and dosage:
Rat is divided into 4 groups at random, 10 every group, is respectively blank group, model group, positive controls (prednisone) and ASC group; The prednisone dosage is 15mg/kg, and ASC organizes to being equivalent to 10g raw medicinal herbs/kg, and it is 10ml/kg that model group gives isopyknic distilled water.
4, experimental technique:
After the anesthesia of lumbar injection pentobarbital sodium, under aseptic condition, with 30 rats underwent left side nephrectomys, postoperative is divided into 3 groups at random, presses 5mg/kg from its tail vein injection doxorubicin hydrochloride after 1 week earlier; In addition to the sham-operation of going ahead of the rest of 10 rats in normal control group, 1 week the back tail vein injection saline.Began gastric infusion the same day in modeling, once a day, continuous 35 days, blank group and model group were irritated stomach and are given the equivalent distilled water.Administration 35d claims each Mus weight, and rat is positioned over collection 24h urinary assay urine protein content (Coomassie brilliant blue method) in the metabolic cage.24h vena orbitalis posterior clump is got blood and measures total protein, creatinine, blood urea nitrogen, T-CHOL, triglyceride in the blood in automatic biochemistry analyzer after the last administration, gets kidney after disconnected neck is put to death and does histological examination (inspection method is the same).
5, statistical procedures: experimental data adopts SPSS 12.0 softwares to handle with average (X) expression, whole experimental datas.
6, experimental result: concrete experimental result sees Table 6-8.
Table 6ASC is to excising the influence (n=10) of mRNA IN ADRIAMYCIN NEPHROPATHY rat model urine albumen amount behind the left kidney
Annotate: P is the probability of comparing with model group
Relatively last table data Chinese milkvetch seed extractive as can be known have the urine protein reducing effect suitable with prednisone to mRNA IN ADRIAMYCIN NEPHROPATHY rat model after excising left kidney, and have statistical significance.
Table 7 blood physicochemical data result (n=10)
Annotate: * and model group compare, p＜0.05
Relatively go up in the blood that the visible Chinese milkvetch seed extractive of table data can make mRNA IN ADRIAMYCIN NEPHROPATHY rat model behind the left kidney of excision total protein content and keep substantially normally, make creatinine Cr and content of triglyceride obtain to a certain degree reduction.
Table 7 histopathologic examination (n=10)
Annotate: * and model group compare, p＜0.05
Relatively go up data in the table and can find that Chinese milkvetch seed extractive can make that the pathological phenomenons such as cellular edema, tubular ectasia and interstitial fibrosis of mRNA IN ADRIAMYCIN NEPHROPATHY rat model are significantly improved behind the left kidney of excision.
Five, extract is to acute toxicity test in mice
1, laboratory animal: healthy ICR mice, male and female half and half, body weight 20 ± 2g is available from Zhejiang Province's Experimental Animal Center.The quality certification number: SCXK (Zhejiang 2003-0001).
2, experiment is subjected to test product and medicine:
Chinese milkvetch seed extractive: n-butyl alcohol extract, ASC.
3, experimental technique:
1) trial test
Before formally testing, at first do trial test, so that provide foundation for formal test.18 mices are divided into three groups at random, n-butyl alcohol extract group, ASC group and blank group.Water 16h is can't help in fasting before irritating stomach.N-butyl alcohol extract group: irritating for 1 time to n-butyl alcohol extract for the Cmax (0.15g/mL) of filling stomach, the maximum volume (every 10g body weight 0.4mL) that mouse stomach can bear; ASC group: irritating for 1 time to ASC for the Cmax (0.15g/mL) of filling stomach, the maximum volume (every 10g body weight 0.4mL) that mouse stomach can bear; The conventional raising observed 3d after the administration.Blank group: irritate stomach equivalent distilled water, method is with the administration group.Mice all survives as a result, therefore, can't measure the LD of this compound preparation
50, point out this extract toxicity very low, be safe, so measure the maximum dosage-feeding of this medicine to mice.
2) mensuration of maximum dosage-feeding
60 mices are divided into 20 of blank groups, 20 of n-butyl alcohol extract groups, 20 of ASC groups, every group of male and female half and half at random.Fasting 16h before the test, normal drinking-water.Every mice of n-butyl alcohol extract group is pressed the n-butyl alcohol extract suspension of the dosage filling stomach 0.15g/mL concentration of 0.4mL/10g; Every mice of ASC group is pressed the ASC suspension of the dosage filling stomach 0.15g/mL concentration of 0.4mL/10g; The blank group is irritated the isopyknic distilled water of stomach, administration every day 2 times, 6h at interval.After the administration, observe 7d, take by weighing its body weight every day and observe reaction of animals.The conventional raising after the administration, natural lighting 12h, room temperature (20～24) ℃, humidity 60%.Test took by weighing body weight on the 8th day after, put to death mice, the tissue of major organs such as its heart of anatomic observation, liver, spleen, lung, kidney changes.
4, experimental result
1) the outward appearance sign no abnormality seen of two groups of mices behind the ordinary circumstance gastric infusion, activity freely, feed, drinking-water, feces are all normal, fur is bright and clean, mouth, nose, eye and Anal cleaning do not have secretions.
2) body weight change situation gastric infusion is 7 days, weight increase, body weight is (20.37 ± 0.51) g before the administration of n-butyl alcohol extract group, body weight is (20.39 ± 0.54) g before the ASC administration, blank group body weight is (20.35 ± 0.62) g, and the 7th day body weight, n-butyl alcohol extract group are (24.73 ± 1.31) g, the ASC group is (24.64 ± 1.28) g, and the blank group is (24.36 ± 1.19) g.N-butyl alcohol extract group and ASC group compare P＞0.05, the equal not statistically significant of difference respectively with blank group body weight.
40 mices that main organs dissection perusal experimentizes all survive, and do not have obvious toxic reaction.Dissect finding: thoracic cavity, abdominal cavity no abnormality seen liquid, intestinal tube is not seen flatulence, important organs such as the heart, liver, spleen, lung, kidney, brain are not seen color, paramophia, there are not petechia or other pathological changes, the gastric mucosa color is ruddy, no petechia or ulcer, and postmortem is no abnormal.
The mice maximum dosage-feeding is the 12g/kg body weight.
N-butyl alcohol extract and ASC do not measure LD for ICR strain mouse stomach
50, its maximum tolerated dose is the 12g/kg body weight, is about clinical patients plan 240 times with dosage, illustrates that this extract does not have the overt toxicity reaction, and is safe and reliable.
Claims (5)
1. the Chinese milkvetch seed extractive purposes in the medicine of preparation treatment nephrotic syndrome, described Chinese milkvetch seed extractive prepares by following steps:
1) Semen astragali sinici parching to brown is pulverized, and crosses 10 mesh sieves, adds 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, and each 2h filters, merging filtrate, decompression recycling ethanol must extract concentrated solution;
2) should extract concentrated solution adsorbs with macroporous adsorptive resins, after washing post with 6 times of water gagings and 7 times of amount 30% ethanol successively, with 9 times of amount 70% ethanol elutions, the eluent decompression and solvent recovery, at 60 ℃ of following vacuum dryings, get dry extract, dry extract is broken into fine powder, just be prepared into extract of the present invention.
2. the Chinese milkvetch seed extractive purposes in the medicine of preparation treatment nephrotic syndrome, described Chinese milkvetch seed extractive prepares by following steps:
1) Semen astragali sinici parching to brown is pulverized, and crosses 10 mesh sieves, adds 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, and each 2h filters, merging filtrate, decompression recycling ethanol must extract concentrated solution;
2) should extract concentrated solution, successively with petroleum ether, the extraction of ethyl acetate equal-volume, after each extracts 3 times,, merge butanol extraction liquid with equal-volume n-butanol extraction 3 times, decompression and solvent recovery, at 60 ℃ of following vacuum dryings, in drying baker, place the residual n-butyl alcohol of activated carbon adsorption, get dry extract, dry extract is broken into fine powder, just has been prepared into extract of the present invention.
3. the preparation method of a Chinese milkvetch seed extractive is characterized in that the step of this method is as follows:
1) Semen astragali sinici parching to brown is pulverized, and crosses 10 mesh sieves, adds 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, and each 2h filters, merging filtrate, decompression recycling ethanol must extract concentrated solution;
2) should extract concentrated solution adsorbs with macroporous adsorptive resins, after washing post with 6 times of water gagings and 7 times of amount 30% ethanol successively, with 9 times of amount 70% ethanol elutions, the eluent decompression and solvent recovery, at 60 ℃ of following vacuum dryings, get dry extract, dry extract is broken into fine powder, just be prepared into extract of the present invention.
4. the preparation method of a kind of Chinese milkvetch seed extractive according to claim 3 is characterized in that: big pore adsorption resin comprises nonpolar, low pole and middle polarity macroporous adsorbent resin described step 2).
5. the preparation method of a Chinese milkvetch seed extractive is characterized in that the step of this method is as follows:
1) Semen astragali sinici parching to brown is pulverized, and crosses 10 mesh sieves, adds 6 times of amount 60% soak with ethanol 2h, reflux, extract, 2 times, and each 2h filters, merging filtrate, decompression recycling ethanol must extract concentrated solution;
2) should extract concentrated solution, successively with petroleum ether, the extraction of ethyl acetate equal-volume, after each extracts 3 times,, merge butanol extraction liquid with equal-volume n-butanol extraction 3 times, decompression and solvent recovery, at 60 ℃ of following vacuum dryings, in drying baker, place the residual n-butyl alcohol of activated carbon adsorption, get dry extract, dry extract is broken into fine powder, just has been prepared into extract of the present invention.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100977218A CN101530458B (en) | 2009-04-16 | 2009-04-16 | Application and preparing method of Chinese milkvetch seed extractive |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009100977218A CN101530458B (en) | 2009-04-16 | 2009-04-16 | Application and preparing method of Chinese milkvetch seed extractive |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101530458A CN101530458A (en) | 2009-09-16 |
| CN101530458B true CN101530458B (en) | 2011-09-21 |
Family
ID=41101453
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2009100977218A Active CN101530458B (en) | 2009-04-16 | 2009-04-16 | Application and preparing method of Chinese milkvetch seed extractive |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101530458B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1634398A (en) * | 2004-11-22 | 2005-07-06 | 华东师范大学 | Method for applying complete puncturevine herb in preparing medicine for treating insulin resisting hyperglycemia and hypertension |
| JP2005220100A (en) * | 2004-02-06 | 2005-08-18 | Maruzen Pharmaceut Co Ltd | Anti-ageing agent, platelet aggregation inhibitor, antioxidant, anti-allergic agent, skin cosmetic, and food and drink |
-
2009
- 2009-04-16 CN CN2009100977218A patent/CN101530458B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005220100A (en) * | 2004-02-06 | 2005-08-18 | Maruzen Pharmaceut Co Ltd | Anti-ageing agent, platelet aggregation inhibitor, antioxidant, anti-allergic agent, skin cosmetic, and food and drink |
| CN1634398A (en) * | 2004-11-22 | 2005-07-06 | 华东师范大学 | Method for applying complete puncturevine herb in preparing medicine for treating insulin resisting hyperglycemia and hypertension |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101530458A (en) | 2009-09-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101161251B (en) | Teasel root total saponin as well as its extracting method and application | |
| CN100500160C (en) | Medicinal composition containing epimedium active constituent and its application | |
| CN101810715A (en) | New application of fructus evodiae and extracts and compounds thereof | |
| CN101011545A (en) | Method for extracting effective sites group of smilax China root | |
| CN101007157A (en) | Traditional Chinese medicine compound preparation for treating chronic cholecystitis | |
| CN105982970A (en) | Traditional Chinese medicinal composition for treating psoriasis and preparation method of traditional Chinese medicinal composition | |
| CN101850094B (en) | Chinese medicinal adhesive plaster | |
| CN101530458B (en) | Application and preparing method of Chinese milkvetch seed extractive | |
| CN101041037A (en) | Drug for curing diabetes and nephropathy and its preparing method | |
| CN109078156A (en) | It is a kind of to treat calculi in urinary system Chinese medicine and preparation method thereof | |
| CN1234407C (en) | Medicine for treating chronic prostatitis and its preparing method | |
| CN101406556B (en) | Chinese medicine preparation for treating acute prostatitis, prostatic hyperplasia and preparation method thereof | |
| CN104998085A (en) | Chinese herbal compound composition capable of enhancing immune function and preparation method thereof | |
| CN100493522C (en) | Medicinal composition of oxymatrine and polysaccharide | |
| CN105434915A (en) | Pharmaceutical composition for treating urolithiasis for dogs and cats and preparation method of pharmaceutical composition | |
| CN101721598A (en) | Traditional Chinese medicine composition for treating liver diseases and preparation method and use thereof | |
| CN100536899C (en) | Chinese medicine for treating and preventing early diabetes and nephropathy | |
| CN112386671B (en) | Traditional Chinese medicine composition for treating interstitial lung disease and application thereof | |
| CN103040939A (en) | Traditional Chinese medicine composition for treating cold, as well as preparation method and application thereof | |
| CN104095912A (en) | Traditional Chinese medicine formula for treating rheumatism bone disease and preparation method of Chinese patent medicine of traditional Chinese medicine formula | |
| CN102258593B (en) | Chinese medicinal composition for adjuvant treatment of tumors and preparation method and application thereof | |
| CN102895348B (en) | Pharmaceutical compositions for treating rheumatic joint disease, preparation method thereof and use thereof | |
| CN1231194A (en) | Capsule for treating pointed condyloma and preparation process | |
| CN101301403B (en) | Clerodendrum bungei extract and use thereof for treating airway hyperreactivity | |
| CN105326879A (en) | Fatty acid extract of rape pollen as well as preparation method and application of fatty acid extract |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 310053 No. 677, bin Kang Road, Hangzhou, Zhejiang, Binjiang District Patentee after: Zhejiang jinshiya Pharmaceutical Technology Co.,Ltd. Address before: 310053 No. 677, bin Kang Road, Hangzhou, Zhejiang, Binjiang District Patentee before: HANGZHOU CHUANGXIN TCM STANDARDIZATION INST. Co.,Ltd. |