CN104189044A - Application of spanishneedles herb extract in preparation of hypoglycemic agent - Google Patents
Application of spanishneedles herb extract in preparation of hypoglycemic agent Download PDFInfo
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- CN104189044A CN104189044A CN201410401686.5A CN201410401686A CN104189044A CN 104189044 A CN104189044 A CN 104189044A CN 201410401686 A CN201410401686 A CN 201410401686A CN 104189044 A CN104189044 A CN 104189044A
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Abstract
The invention relates to application of spanishneedles herb extract in preparation of a hypoglycemic agent, effectively solving a problem of hypoglycemic agent preparation. According to the application of the spanishneedles herb extract in preparation of the hypoglycemic agent, the spanishneedles herb extract is obtained by the following steps: taking dried spanishneedles herb, crushing the spanishneedles herb into crude powder, adding the crude powder into alcohol for soaking, refluxing and extracting twice, filtering, combining filtrate, recycling alcohol under reduced pressure until alcohol smell does not exist, concentrating the filtrate into concentrated liquor with relative density of 1.10-1.20 at 50 DEG C, adding distilled water into the concentrated liquor for dispersing the concentrated liquor into a dispersion liquid; placing the dispersion liquid onto a macroporous adsorption resin column, eluting by distilled water, discarding eluant, further eluting by 10% alcohol, discarding the eluant and then eluting with 70% alcohol, collecting the eluant, recycling alcohol under reduced pressure, drying and pulverizing to obtain the spanishneedles herb extract, wherein the content of the general flavones of the spanishneedles herb is over 55%. According to the invention, the raw materials are rich, the preparation method is simple, and the prepared products have hypoglycemic effects and can be used for preparing the hypoglycemic agent, so that the application of the spanishneedles herb extract in preparation of the hypoglycemic agent is realized.
Description
Technical field
The present invention relates to medicine, the particularly application of a kind of Herba Bidentis Bipinnatae extract in preparing hypoglycemic medicine.
Background technology
One of Herba Bidentis Bipinnatae, first appeared in < < Bencao Shiyi > >, is Compositae annual herb plant.Different name there's something fishy hairpin grass (< < Bencao Shiyi > >), SANYE Bidens bipinnata, bidens pilosa, terrible Hemerocallis citrina Baroni, Shandong lao-ya-tsao, Herba Bidentis Bipinnatae, terrible spicule, fork woman, terrible Fructus Tribuli, sticky body grass, steel fork pin, terrible muscle grass etc.Herba Bidentis Bipinnatae is distributed in the subtropical and tropical zones in Asia and America, and the East China, Central China, south China of China, each provinces and regions, southwest, grows in wasteland, roadside, hillside and the field of height above sea level 50-3100 rice, is the conventional medical herbs unisexuality gymnosperm of folks of china.Hardship, flat.Heat-clearing and toxic substances removing, promoting blood circulation by removing wind.For laryngopharynx swelling and pain, vomit and diarrhoea, dyspepsia, rheumatic arthritis, malaria, furuncle, venom, treating swelling and pain by traumatic injury.But the open report of the application so far there are no Herba Bidentis Bipinnatae extract in preparing hypoglycemic medicine.
Summary of the invention
For above-mentioned situation, for overcoming the defect of prior art, the present invention's object is just to provide the application of a kind of Herba Bidentis Bipinnatae extract in preparing hypoglycemic medicine, can effectively solve the problem of preparing hypoglycemic medicine.
The technical scheme that the present invention solves is, the application of Herba Bidentis Bipinnatae extract in preparing hypoglycemic medicine, this Herba Bidentis Bipinnatae extract is to get dry Herba Bidentis Bipinnatae herb, be ground into coarse powder, add soak with ethanol 2h, reflux, extract, 2 times, filters, merging filtrate, decompression recycling ethanol is to without alcohol taste, and is concentrated into the concentrated solution that 50 ℃ of relative densities are 1.10～1.20, and concentrated solution adding distil water is dispersed into dispersion liquid; Macroporous adsorptive resins on dispersion liquid, first use distilled water eluting, discard distilled water eluent, continue with the ethanol elution of mass concentration 10%, discard the ethanol elution of mass concentration 10%, use again the ethanol elution of mass concentration 70%, collect the ethanol elution of mass concentration 70%, decompression recycling ethanol, drying and crushing, obtain Herba Bidentis Bipinnatae extract, wherein Sticktight general flavone content is more than 55%.
Abundant raw material of the present invention, preparation method is simple, and gained prepared product has blood sugar reducing function, can be used for preparing hypoglycemic medicine, realize the application of Herba Bidentis Bipinnatae extract in preparing hypoglycemic medicine, and then improve the medical value of Herba Bidentis Bipinnatae and the new purposes of developing Herba Bidentis Bipinnatae, have good economic and social benefit.
The specific embodiment
Below in conjunction with concrete condition, the specific embodiment of the present invention is elaborated.
The application of Herba Bidentis Bipinnatae extract of the present invention in preparing hypoglycemic medicine, this Herba Bidentis Bipinnatae extract is to get dry Herba Bidentis Bipinnatae herb, pulverizing was the coarse powder of 20 mesh sieves, the soak with ethanol 2h that at every turn adds 10 times of weight mass concentrations 80%, reflux, extract, 2 times, each 1h, filter, merging filtrate, decompression recycling ethanol is to without alcohol taste, and is concentrated into the concentrated solution that 50 ℃ of relative densities are 1.10～1.20, concentrated solution adding distil water is dispersed to and is equivalent to obtain dispersion liquid containing crude drug amount 0.3g/ml, and adjusting pH is 4.0; AB-8 type macroporous adsorptive resins on dispersion liquid, first use 4 times of column volume distilled water eluting, discard distilled water eluent, continue with the ethanol elution of the mass concentration 10% of 4 times of column volumes, discard the ethanol elution of mass concentration 10%, use again the ethanol elution of the mass concentration 70% of 8 times of column volumes, collect the ethanol elution of mass concentration 70%, decompression recycling ethanol, 50 ℃ of drying and crushing, obtain Herba Bidentis Bipinnatae extract, wherein Sticktight general flavone content is more than 55%.
The existing blood sugar reducing function of Herba Bidentis Bipinnatae extract of the present invention, can be effective to prepare hypoglycemic medicine, and and then can prepare treatment diabetes medicament, and through test obtained sufficient proof, related tests data is as follows:
One, Herba Bidentis Bipinnatae extract causes the impact of mice hyperglycemia model on epinephrine
1. experiment material
1.1 experiment reagent
Herba Bidentis Bipinnatae extract of the present invention; Metformin hydrochloride tablet, commercially available; Normal saline, Henan Shuan Hehuali pharmaceutcal corporation, Ltd; Adrenalin hydrochloride, Shanghai Hefeng Pharmaceutical Co., Ltd.; Sodium carboxymethyl cellulose, Tianjin Heng Xing chemical reagent Manufacturing Co., Ltd; Glucose assays test kit, is produced and is provided by Zhongsheng Beikong Biological Science & Technology Co., Ltd.; Hepatic glycogen is measured test kit, builds up Bioengineering Research Institute produce by Nanjing; Concentrated sulphuric acid, Shanghai Zhen Qi chemical reagent company limited.
1.2 experimental apparatus
AL204 series electronic balance, prunus mume (sieb.) sieb.et zucc. Teller-holder benefit instrument (Shanghai) Co., Ltd.; VIS-7220N visible spectrophotometer: Beijing Rayleigh Analytical Instrument Co., Ltd; TGL-16G desk centrifuge, Anting Scientific Instrument Factory, Shanghai produces; HHS type electric-heated thermostatic water bath, Shanghai Medical Equipment Plant of Bo Xun Industrial Co., Ltd.; KDC-160HR High speed refrigerated centrifuge, Zhong Jia branch company of Keda Innovation Co., Ltd produces; Adjustable liquid-transfering gun, Shanghai Lei Bo Analytical Instrument Co., Ltd.
1.3 laboratory animal
Kunming mouse, male, body weight 18～22g, is provided by Lukang Medical Co., Ltd., Shandong; The quality certification number: 0014964; Laboratory quality certification SYXK (Henan) 2010-001.
2. experimental technique
2.1 modelings and administration
72 of Kunming mouses getting body weight and be 18-22g, male, adaptability is evenly divided into 6 groups at random according to body weight after raising, and is respectively blank group, model group, metformin group, the large, medium and small dosage group of Herba Bidentis Bipinnatae extract, 12 every group.According to group, gavage respectively relative medicine, blank group and model group gavage isopyknic 0.5%CMC-Na (0.1ml/10g), metformin hydrochloride tablet (375mg/kg, for 15 times of clinical dosage, CMC-Na preparation with 0.5%, 0.1ml/10g), Herba Bidentis Bipinnatae extract is large, in, small dose group (400, 200, 100mg/kg, CMC-Na preparation with 0.5%, 0.1ml/10g), successive administration 7 days, all lumbar injection adrenalin hydrochloride (0.2mg/kg) modelings except blank group after last administration 2h, the normal saline of blank group injection equivalent, respectively at 30min after modeling, during 60min, get blood, centrifugal after blood coagulation, get serum, press the description of glucose sugar test kit and survey blood glucose value.60min dissects mice after getting blood, gets the liver of suitable weight, by hepatic glycogen test kit description, measures hepatic glycogen content in liver.
3. statistical method
Data analysis carries out the statistical procedures of data information, average ± standard deviation for measurement data with the medical statistics bag of SPSS 17.0for windows
represent, relatively adopt one factor analysis of variance between each group, the neat person of variance test adopts Ridit check with least significant difference (LSD) method, heterogeneity of variance person with Games-Howell method check, ranked data.
4. experimental result and brief summary
Table 1 Herba Bidentis Bipinnatae extract causes the impact (x ± s) of diabetes mice model blood glucose, hepatic glycogen on epinephrine
* represent that with model group, than P<0.05, * * represents to compare P<0.01 with model group
From upper figure, to compare for blank group, the blood glucose value of model group significantly raises (P<0.01), and hepatic glycogen significantly reduces (P<0.01), and model copy success is described.Compare with model group, Herba Bidentis Bipinnatae extract small dose group can obviously reduce the blood glucose value (P<0.05) of 30 minutes, 60 minutes, metformin hydrochloride group, the big or middle dosage group of Herba Bidentis Bipinnatae extract all can remarkable 60 minutes blood glucose value (P<0.01), each medication group of Herba Bidentis Bipinnatae extract hepatic glycogen content (P<0.01) that all can significantly raise.The blood sugar reducing function that each dosage group of Herba Bidentis Bipinnatae extract is described is relevant with the decomposition of hepatic glycogen.
Two, Herba Bidentis Bipinnatae extract causes the impact of diabetes mice model on streptozotocin
1. experiment material
1.1 experiment reagent
Herba Bidentis Bipinnatae extract of the present invention; Metformin hydrochloride tablet, commercially available; Normal saline, Henan Shuan Hehuali pharmaceutcal corporation, Ltd; Sodium carboxymethyl cellulose, Tianjin Heng Xing chemical reagent Manufacturing Co., Ltd; Streptozotocin, sigma company provides; Citric acid, Tianjin Zhi Yuan chemical reagent company limited; Sodium citrate, Tianjin Zhi Yuan chemical reagent company limited; Glucose assays test kit, is produced and is provided by Zhongsheng Beikong Biological Science & Technology Co., Ltd.; Hepatic glycogen is measured test kit, builds up Bioengineering Research Institute produce by Nanjing; Insulin, R & D company produces; Glycated serum protein, builds up Bioengineering Research Institute by Nanjing and produces; Formaldehyde, Yantai City is Chemical Co., Ltd. in pairs; Concentrated sulphuric acid, Shanghai Zhen Qi chemical reagent company limited.
1.2 experimental apparatus
AL204 series electronic balance, prunus mume (sieb.) sieb.et zucc. Teller-holder benefit instrument (Shanghai) Co., Ltd.; VIS-7220N visible spectrophotometer, Beijing Rayleigh Analytical Instrument Co., Ltd; TGL-16G desk centrifuge, Anting Scientific Instrument Factory, Shanghai produces; HHS type electric-heated thermostatic water bath, Shanghai Medical Equipment Plant of Bo Xun Industrial Co., Ltd.; KDC-160HR High speed refrigerated centrifuge, Zhong Jia branch company of Keda Innovation Co., Ltd produces; TES-1380PH acidometer, TES Electronic Industrial Corporation; Microplate reader, U.S. BIO-RAD company, model 680; Adjustable liquid-transfering gun, Shanghai Lei Bo Analytical Instrument Co., Ltd; OLYMPUS BX61 electric microscope, Japanese OLYMPUS company produces; TES-1380PH acidometer, TES Electronic Industrial Corporation.
1.3 laboratory animal
Kunming mouse, male, body weight 18～22g, is provided by Lukang Medical Co., Ltd., Shandong, the quality certification number: 0017170; Laboratory quality certification SYXK (Henan) 2010-001.
2. experimental technique
2.1 modelings and administration
Get 108 of the Kunming mouses of body weight 18-22g, male, after adaptability is raised, according to blank group of 12 conducts of body weight random choose, all the other are modeling group, modeling group before experiment starts, weigh after the streptozotocin 80mg/kg that is mixed with of tail vein injection citric acid-sodium citrate buffer, after 7 days, afterbody blood sampling, measures blood glucose, choose 16.7mmol/L< blood glucose value <21mmol/L, and obviously there is 60 of the mices of polydipsia polyphagia polyuria symptom.Be divided at random 5 groups, be respectively model group, metformin group, the large, medium and small dosage group of Herba Bidentis Bipinnatae extract, 12 every group.According to group, gavage respectively relative medicine, blank group and model group gavage the 0.5%CMC-Na (0.1ml/10g) of same volume, metformin hydrochloride tablet group (375mg/kg, for 15 times of clinical dosage, CMC-Na preparation with 0.5%, 0.1ml/10g), the large, medium and small dosage group of Herba Bidentis Bipinnatae extract (400,200,100mg/kg, CMC-Na preparation with 0.5%, 0.1ml/10g), once a day, successive administration 30 days, during respectively at after administration the 10th, 20,30 days, afterbody blood sampling, measures blood glucose value.In the 31st day, 2h after gavage, got blood, separation of serum, and the content of survey serum insulin, glycation serum protein, gets liver appropriate, surveys hepatic glycogen content; Get pancreas, kidney formaldehyde is fixed, and does tissue pathological slice, observes Histopathologic change.
The preparation of citric acid-sodium citrate buffer: take respectively 500.4mg citric acid and 501.2 sodium citrates and be dissolved in the volumetric flask of 50ml, this solution is the citric acid-sodium citrate buffer of ph=4.2.For guaranteeing its accuracy, can proofread and correct with PH meter.Preparation before use (because streptozotocin is very unstable, and larger to vascular stimulation, with buffer preparation streptozotocin, can alleviate these negative effects
3. statistical method
Data analysis carries out the statistical procedures of data information with the medical statistics bag of SPSS 17.0for windows, average ± standard deviation for measurement data (x ± s) represents, between each group, relatively adopt one factor analysis of variance, the neat person of variance test uses least significant difference (LSD) method, heterogeneity of variance person adopts Ridit check with Games-Howell method check, ranked data.
Experimental result and brief summary
Table 2 Herba Bidentis Bipinnatae extract causes the impact (x ± s) of diabetes mice model blood glucose on streptozotocin
* represent that with model group, than P<0.05, * * represents to compare P<0.01 with model group
As can be seen from Table 2, compare with blank group, the equal significance of the blood glucose of model group in the time of the 10th, 20,30 days raises (P<0.01), and model copy success is described.Compare with model group, metformin group can significantly reduce (P<0.01) mice blood glucose value of the 10th, 20,30 days, and the large, medium and small dosage group of Herba Bidentis Bipinnatae extract can significantly reduce (P<0.01) mice blood glucose value of the 20th, 30 days.Illustrate that Herba Bidentis Bipinnatae extract can effectively reduce the blood glucose value of model.
The impact of table 3 Herba Bidentis Bipinnatae extract on diabetes model hepatic glycogen, insulin, glycated serum protein
* represent that with model group, than P<0.05, * * represents to compare P<0.01 with model group
As can be seen from Table 3: compare with blank group, model group hepatic glycogen content and insulin level all significantly reduce (P<0.01), glycated serum protein content significantly raises (P<0.01), and model copy success is described.Compare with model group, metformin group (P<0.01) hepatic glycogen content that significantly raises that can significantly raise, significantly reduces the content of (P<0.01) glycated serum protein; The big or middle dosage group of Herba Bidentis Bipinnatae extract all can significantly raise (P<0.01) hepatic glycogen content and insulin level, significantly reduce the content of (P<0.01) glycated serum protein, Herba Bidentis Bipinnatae extract small dose group can significantly raise (P<0.01) hepatic glycogen content and insulin level, can obviously reduce the content of (P<0.05) glycated serum protein.
The impact of table 4 Herba Bidentis Bipinnatae extract on streptozotocin induced mice diabetes model pancreatic tissue
In "-" pancreatic tissue, islets of langerhans is more than 4, and islet cells endochylema is abundant, and nucleus is loose; In "+" pancreatic tissue, islets of langerhans is more than 4, the atrophy of minority islet cells endochylema, and a few cell core is intensive; In " ++ " pancreatic tissue, islets of langerhans is individual at 2-3, the atrophy of part islet cells endochylema, and parts of fine karyon is intensive; In " +++ " pancreatic tissue, islets of langerhans is at 0-1, and all islet cells endochylema almost disappears, and all nucleus is intensive
By upper table and pathology photo, can be found out, blank group Mice Islet Cells is more open, sharpness of border, cytoplasm is compared with horn of plenty (P<0.01), the islet cells that model group occurs reduces, cytoplasm atrophy, some atrophy even completely, illustrates that the islet cells of model group animal is destroyed.Compare with model group, metformin group islet cells endochylema significantly improves, and the large, medium and small dosage group of Herba Bidentis Bipinnatae extract all can alleviate the atrophy of islet cells, and the intensive pathological change (P<0.01) of nucleus.
The impact of table 5 Herba Bidentis Bipinnatae extract on streptozotocin induced mice diabetes model nephridial tissue
"-" messangial cell is normal, and renal capsule is normal; The slightly atrophy of "+" messangial cell, renal capsule is slightly expanded; The atrophy of " ++ " messangial cell, capsular space expansion; The obvious atrophy of " +++ " messangial cell, capsular space is obviously expanded
From upper table and pathology photo, can find out, blank group glomerule, renal tubules is normal, and renal cells edema and the interstitial of model group mice have cell infiltration in various degree.Through Radit check, illustrate that model group mouse kidney damages.Compare with model group, metformin group, the heavy dose of group of Herba Bidentis Bipinnatae extract, middle dosage group, the slightly atrophy of small dose group glomerule, renal cells inflammation alleviates.
Test fully proves that Herba Bidentis Bipinnatae extract has blood sugar reducing function, be effective to prepare hypoglycemic medicine, and through diabetics 105 examples are carried out to clinical practice, take every day 3 times, each 0.5-1g, statistical result after 15 days, result shows, except 1 example finds no diabetes blood glucose unchanged rear (but also not increasing the weight of), other all have blood glucose in various degree to reduce and symptom on improvement, effective percentage is up to more than 99%, thereby prove that Herba Bidentis Bipinnatae extract of the present invention can be used for the medicine of preparation treatment diabetes, solve the medication problem for the treatment of diabetes, opened up the new purposes of Herba Bidentis Bipinnatae, there is huge economy and social meaning.
Claims (1)
1. the application of Herba Bidentis Bipinnatae extract in preparing hypoglycemic medicine, this Herba Bidentis Bipinnatae extract is to get dry Herba Bidentis Bipinnatae herb, pulverizing was the coarse powder of 20 mesh sieves, the soak with ethanol 2h that at every turn adds 10 times of weight mass concentrations 80%, reflux, extract, 2 times, each 1h, filter, merging filtrate, decompression recycling ethanol is to without alcohol taste, and is concentrated into the concentrated solution that 50 ℃ of relative densities are 1.10～1.20, concentrated solution adding distil water is dispersed to and is equivalent to obtain dispersion liquid containing crude drug amount 0.3g/ml, and adjusting pH is 4.0; AB-8 type macroporous adsorptive resins on dispersion liquid, first use 4 times of column volume distilled water eluting, discard distilled water eluent, continue with the ethanol elution of the mass concentration 10% of 4 times of column volumes, discard the ethanol elution of mass concentration 10%, use again the ethanol elution of the mass concentration 70% of 8 times of column volumes, collect the ethanol elution of mass concentration 70%, decompression recycling ethanol, 50 ℃ of drying and crushing, obtain Herba Bidentis Bipinnatae extract, wherein Sticktight general flavone content is more than 55%.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106389497A (en) * | 2016-11-24 | 2017-02-15 | 广东省农业科学院动物科学研究所 | Preparation method of sticktight soluble granules |
| CN111870649A (en) * | 2020-09-04 | 2020-11-03 | 杨鹏 | Traditional Chinese medicine composition for treating type II diabetes and preparation method thereof |
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| CN103652152A (en) * | 2012-09-26 | 2014-03-26 | 陈思宏 | Spanishneedles herb healthcare tea and preparation method thereof |
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- 2014-08-13 CN CN201410401686.5A patent/CN104189044A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103652152A (en) * | 2012-09-26 | 2014-03-26 | 陈思宏 | Spanishneedles herb healthcare tea and preparation method thereof |
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| 曲永胜: "鬼针草黄酮化合物的提取及工艺优化", 《山东医药》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106389497A (en) * | 2016-11-24 | 2017-02-15 | 广东省农业科学院动物科学研究所 | Preparation method of sticktight soluble granules |
| CN106389497B (en) * | 2016-11-24 | 2020-02-18 | 广东省农业科学院动物科学研究所 | Preparation method of bidens bipinnata soluble particles |
| CN111870649A (en) * | 2020-09-04 | 2020-11-03 | 杨鹏 | Traditional Chinese medicine composition for treating type II diabetes and preparation method thereof |
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