CN101229316B - Rhizoma anemarrhenae extrac - Google Patents
Rhizoma anemarrhenae extrac Download PDFInfo
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- CN101229316B CN101229316B CN2008100262051A CN200810026205A CN101229316B CN 101229316 B CN101229316 B CN 101229316B CN 2008100262051 A CN2008100262051 A CN 2008100262051A CN 200810026205 A CN200810026205 A CN 200810026205A CN 101229316 B CN101229316 B CN 101229316B
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Abstract
The invention discloses an anemarrhenae extract and the application as a medicine treating type II diabetes thereof. The invention uses membrane separation technology, macropore to absorb resin separation and refines anemarrhenae extract after extracting the hizome anemarrhenae by water and ethanol with water. The main activities of the anemarrhenae extract of the invention are three kinds or any two kinds among anemarrhenae saponins, anemarrhenae polysaccharides and double benzene pyrrole ketones, and the total content of the main active components is up to more than 50 percent. The pharmacological experiment of the anemarrhenae extract of the invention proves that the medicine has a big medical value and economic benefit as the medicine for treating or preventing type II diabetes and the complications thereof.
Description
Technical field
The present invention relates to the diabetes medicament field, be specifically related to a kind of Rhizoma Anemarrhenae extract that is used for the treatment of diabetes.
Background technology
The Rhizoma Anemarrhenae (Rhizoma Anemarrhenae) is the dry rhizome of liliaceous plant (Anemarrhenaasphodeloides), its nature and flavor bitter cold, and the effect that clearing away heat-fire is arranged, promote the production of body fluid and moisturize is a clearing up internal heat by using drugs of bitter in taste and cold in nature medicine commonly used.
Modern study shows, the timosaponin that contains in the Rhizoma Anemarrhenae (timosaponin), chimonin (chimonin, mangiferin) and glucoside and anemaran (anemaran) from different machine-processed blood sugar lowering, can be used for treating non-insulin-dependent diabetes mellitus.
Timosaponin can strengthen the sensitivity of peripheral tissues to insulin by recovering the beta Cell of islet function, suppresses the control of three kinds of approach realizations of alpha-glucosidase activity to blood sugar level.
Anemaran can reduce normal mouse and by the blood glucose target of alloxan type mice.The hypoglycemic activity of anemaran is synthetic with its increase liver glycogen, the minimizing liver glycogen decomposes, it is relevant to factors such as glucose uptakes to increase skeletal muscle.In addition, aldose reductase (AR) plays pivotal role in the morbidity of sugared cataract, and also cornea, optic nerve and the peripheroneural pathological changes that causes with diabetes is relevant.Anemaran can be used as aldose reductase (AR) inhibitor, the syndrome of diabetes-alleviating.
Chimonin and chimonin-7-O-β-glucosides does not have influence to normal mouse blood sugar, but can reduce the KK-Ay mouse blood sugar and the trend that reduces serum insulin levels is arranged, and its blood sugar lowering mechanism may be to reduce insulin resistance.And chimonin is in type 2 diabetes mellitus, and is useful and can be used as the treatment that anticoagulant is used for myocardial infarction to hyperlipemia.
Summary of the invention
The object of the present invention is to provide a kind of Rhizoma Anemarrhenae extract that type 2 diabetes mellitus is had therapeutical effect.
Another object of the present invention is to provide the application of above-mentioned Rhizoma Anemarrhenae extract in preparation treatment or prevention type 2 diabetes mellitus medicine.
Rhizoma Anemarrhenae extract of the present invention is prepared from by following method:
(1) after the Rhizoma Anemarrhenae is pulverized, extract 1～3 time with 6～12 times of volume 10%～90% alcoholic solution, each 1～3h is put cold after-filtration, merging filtrate, concentrated thick paste 1; (10%～90% alcoholic solution is meant percentage by volume, and following alcoholic solution all is meant percentage by volume)
(2) get thick paste 1, with 6～12 times of volume water dissolutioies, last macroporous resin column with 3～5 column volume distilled water eluting, discards eluent; With 4～8 column volumes of 40%～80% alcoholic solution eluting, eluent is concentrated into does not have alcohol, filters, and will precipitate drying and crushing, gets extract 1, is Rhizoma Anemarrhenae extract.
In order to improve extraction efficiency, in above-mentioned Rhizoma Anemarrhenae extract preparation method, the residue after the alcohol extraction further can be used water extraction, concrete steps are as follows:
(1) after the Rhizoma Anemarrhenae is pulverized, extract 1～3 time with 6～12 times of volume 10%～90% alcoholic solution, each 1～3h is put cold after-filtration, merging filtrate, concentrated thick paste 1; Filter the back residue and add 6～12 times of volume water extraction 1～3 time, each 1～3h, filtered while hot, merging filtrate, concentrate thick paste 2;
(2) get thick paste 1, with 6～12 times of volume water dissolutioies, last macroporous resin column with 3～5 column volume distilled water eluting, discards eluent; With 4～8 column volumes of 40%～80% alcoholic solution eluting, eluent is concentrated into does not have alcohol, filters, and will precipitate drying and crushing, gets extract 1;
(3) get thick paste 2, add 10～20 times of volume water dissolutioies, trapped fluid is got in ultrafiltration, and concentrate drying gets extract 2; Described ultrafiltration is to adopt the ultrafilter membrane of ultrafilter membrane device and molecular cut off 10000～300000 to carry out ultrafiltration;
(4) the united extraction thing 1,2, promptly get Rhizoma Anemarrhenae extract.
The Rhizoma Anemarrhenae extract 1 that above-mentioned preparation method obtains mainly contain saponins and two benzene pyrrone two constituents, and content is greater than 50%.Rhizoma Anemarrhenae extract 2 mainly contain the anemaran constituents, and content is greater than 50%.
Rhizoma Anemarrhenae extract of the present invention mainly contains at least two classes in following three constituents, this three constituents comprises: saponinss such as 1-timosaponin A-1 I, 1-timosaponin A-1 II, 1-timosaponin A-1 III, 1-timosaponin A-1 IV, timosaponin BI, timosaponin BII, timosaponin BHI, polysaccharide and two benzene pyrrones such as chimonin and chimonin-7-O-β-glucosides such as anemaran A, B, C, D, and these effective ingredient total contents are between 50% and 100%.
Rhizoma Anemarrhenae extract of the present invention can be made into the medicine of various dosage forms such as tablet, capsule, granule, slow releasing agent, injection, is used for treatment or prevent diabetes and complication thereof.Confirm through zoopery, this Rhizoma Anemarrhenae extract has the activity that suppresses the small intestine epithelium alpha-glucosidase, increases target cell Insulin receptor INSR number, improves target cell to insulin sensitivity, alleviate insulin resistant, promote effects such as insulin secretion and blood sugar lowering.
Compared with prior art, the present invention has following beneficial effect: Rhizoma Anemarrhenae extract of the present invention increases serum insulin concentration by protection, reparation pancreatic islet endocrine, and increase liver glycogen content, the activity that suppresses the small intestine epithelium alpha-glucosidase competitively, thereby delay the absorption of intestinal, play the effect of blood sugar lowering sugar; By increasing target cell Insulin receptor INSR number, improve target cell to insulin sensitivity, thereby alleviate insulin resistant; And the diabetic complication to cataract, hyperlipidemia and cardiovascular aspect has preventive and therapeutic effect, therefore can be prepared into the medicine of treatment or prevention type 2 diabetes mellitus and complication thereof, has very big pharmaceutical value and economic benefit.
Description of drawings
Fig. 1 is the curve chart of Rhizoma Anemarrhenae extract to the alpha-glucosidase activity influence.
The specific embodiment
Embodiment 1 preparation Rhizoma Anemarrhenae metformin
(1) rhizoma ane marrhenae is pulverized, with 80% ethanol extraction of 8 times of volumes 2 times, each 1h is put cold after-filtration, merging filtrate, concentrated thick paste 1.
(2) get thick paste 1,,, discard eluent with 5 column volume distilled water eluting with 8 times of volume water dissolutioies, last macroporous resin column.With 8 column volumes of 50% alcoholic solution eluting, eluent is concentrated into does not have alcohol, leaves standstill, and separates out precipitation, filters, and will precipitate drying and crushing, gets extract 1, i.e. Rhizoma Anemarrhenae extract.
(3) take by weighing the above-mentioned Rhizoma Anemarrhenae extract of 250g, add 250g calcium sulfate, 10g polyvinylpolypyrrolidone mixing.Make soft material with an amount of 80% ethanol as wetting agent, granulate with 14 mesh sieves, 12 mesh sieve granulate are crossed in dry back under 60 ℃ of temperature, behind adding 10g polyvinylpolypyrrolidone and the 3g magnesium stearate mixing, make 1000.
Embodiment 2 preparation Rhizoma Anemarrhenae metformins
(1) rhizoma ane marrhenae is pulverized, with 8 times of volumes, 80% ethanol extraction 2 times, each 1h is put cold after-filtration, merging filtrate, concentrate thick paste 1; Filter the back residue and add 8 times of volume water extraction 2 times, each 1, filtered while hot, merging filtrate, concentrate thick paste 2.
(2) get thick paste 1, with 8 times of volume water dissolutioies, last macroporous resin column with 5 column volume distilled water eluting, discards eluent.With 8 column volumes of 50% ethanol elution, eluent is concentrated into does not have alcohol, leaves standstill, and separates out precipitation, filters, and will precipitate drying and crushing, gets extract 1.
(3) get thick paste 2, add 20 times of volume water dissolutioies, the ultrafilter membrane of using ultrafilter membrane device and molecular cut off 300000 filters, and gets trapped fluid, and concentrate drying gets extract 2.
(4) united extraction thing 1 and extract 2 promptly get Rhizoma Anemarrhenae extract.
(5) take by weighing the above-mentioned Rhizoma Anemarrhenae extract of 250g, add 250g calcium sulfate, 10g polyvinylpolypyrrolidone mixing.Make soft material with an amount of 80% ethanol as wetting agent, granulate with 14 mesh sieves, 12 mesh sieve granulate are crossed in dry back under 60 ℃ of temperature, behind adding 10g polyvinylpolypyrrolidone and the 3g magnesium stearate mixing, make 1000.
Embodiment 3 preparation Rhizoma Anemarrhenae Jiangtang capsules
(1) rhizoma ane marrhenae is pulverized, with 10 times of volumes, 80% ethanol extraction 2 times, each 1h is put cold after-filtration, merging filtrate, concentrate thick paste 1; Filter the back residue and add 10 times of volume water extraction 2 times, each 1, filtered while hot, merging filtrate, concentrate thick paste 2.
(2) get thick paste 1, dissolve fully with 8 times of volume water gagings, on the macroporous resin column handled well.With 3 column volume distilled water eluting, discard eluent.With 6 column volumes of 60% ethanol elution, eluent is concentrated into does not have alcohol, leaves standstill, and separates out precipitation, filters, and will precipitate drying and crushing, gets extract 1.
(3) get thick paste 2, add 10 times of volume water dissolutioies, after the ultrafilter membrane of application ultrafilter membrane device and molecular cut off 100000 filters, get trapped fluid, concentrate drying gets extract 2.
(4) united extraction thing 1 and extract 2 promptly get Rhizoma Anemarrhenae extract.
(5) take by weighing the above-mentioned Rhizoma Anemarrhenae extract of 250g, add 250g calcium sulfate, 10g polyvinylpolypyrrolidone mixing.Make soft material with an amount of 80% ethanol as wetting agent, granulate with 14 mesh sieves, 12 mesh sieve granulate are crossed in dry back under 60 ℃ of temperature, insert in the capsulae vacuus, promptly.
Embodiment 4 Rhizoma Anemarrhenae extracts are to the influence of glucose induced hyperglycemia mice blood glucose
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), glibenclamide (Three Jins, Shanxi pharmaceutcal corporation, Ltd, lot number 20021016), glucose injection (Dongya Medicine Co., Ltd., Jiangxi Prov., lot number 2003092732), blood sugar detection test kit (Great Wall, Baoding clinical reagent company, lot number 991514).
2, method
Get 80 of healthy mices, male and female half and half are divided into normal control group, model group, glibenclamide group, the basic, normal, high dosage group of Rhizoma Anemarrhenae extract at random and (are respectively 50,100,150mg/kg).Continuous gastric infusion 7d, normal control group and model group wait the dosage normal saline, glibenclamide group jar stomach 50mg/kg.Fasting 3h before the last administration, 30min lumbar injection glucose injection after the administration, dosage is 2g/kg, causes hyperglycemia model, the 30min posterior orbit is got blood, uses the determination of glucose oxidase blood glucose value, the results are shown in Table 1.
Table 1 Rhizoma Anemarrhenae extract is to the influence of glucose induced hyperglycemia mice blood glucose
| Group | Dosage (mg/Kg) | Blood glucose (mmol/L) |
| Normal control group model matched group Rhizoma Anemarrhenae extract group glibenclamide group | - - 50 100 150 50 | 5.30±0.94 * 11.81±1.03 7.65±1.77 ** 7.32±2.87 ** 6.61±1.11 ** 8.91±2.84 * |
Annotate: compare with model control group,
*P＜0.01,
*P＜0.001.
Embodiment 5 Rhizoma Anemarrhenae extracts are to the influence of glucose induced hyperglycemia mice blood glucose
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), glibenclamide (Three Jins, Shanxi pharmaceutcal corporation, Ltd, lot number 20021016), glucose injection (Dongya Medicine Co., Ltd., Jiangxi Prov., lot number 2003092732), blood sugar detection test kit (Great Wall, Baoding clinical reagent company, lot number 991514).
2, method
Get 80 of healthy mices, male and female half and half are divided into normal control group, model group, glibenclamide group, the basic, normal, high dosage group of Rhizoma Anemarrhenae extract at random and (are respectively 50,100,150mg/kg).Continuous gastric infusion 7d, normal control group and model group wait the dosage normal saline, glibenclamide group jar stomach 50mg/kg.Fasting 3h before the last administration, 30min lumbar injection glucose injection after the administration, dosage is 2g/kg, causes hyperglycemia model, the 30min posterior orbit is got blood, uses the determination of glucose oxidase blood glucose value, the results are shown in Table 2.
Table 2 Rhizoma Anemarrhenae extract is to the influence of glucose induced hyperglycemia mice blood glucose
| Group | Dosage (mg/Kg) | Blood glucose (mmol/L) |
| Normal control group model matched group Rhizoma Anemarrhenae extract group glibenclamide group | - - 50 100 150 50 | 5.30±0.94 * 11.81±1.03 6.94±1.52 ** 6.31±2.14 ** 6.01±1.09 ** 8.91±2.84 * |
Annotate: compare with model control group,
*P＜0.01,
*P＜0.001.
Embodiment 6 Rhizoma Anemarrhenae extracts are to the influence of the blood glucose of diabetic mice due to the streptozotocin
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), (Lik-Sang pharmaceutical factory in Tianjin produces glyburide, lot number 990203), streptozotocin (production of Sigma company), blood sugar detection test kit (Great Wall, Baoding clinical reagent company, lot number 991516).
2, method
Get 80 of healthy male mices, 10 of picked at random are as the normal control group, all the other 70 fasting 12h, press the dosage lumbar injection streptozotocin of 50mg/kg body weight, normal control group injection citrate buffer solution, 1 week was measured fasting blood sugar after modeling, and the mice that the screening blood glucose value surpasses 13mmol/L is the experimental model Mus.50 diabetes experimental model Mus are divided into diabetic model group at random, glyburide (irritating stomach 20mg/kg), the basic, normal, high dosage group of Rhizoma Anemarrhenae extract (is respectively 50,100,150mg/kg).Normal control group and diabetic model group are irritated stomach normal saline, 3 weeks of successive administration.Fasting 12h after the last administration, eye socket is got blood, surveys the change of blood sugar value, the results are shown in Table 3.
Table 3 Rhizoma Anemarrhenae extract is to the influence of the blood glucose of diabetic mice due to the streptozotocin
| Group | Dosage (mg/Kg) | Blood glucose (mmol/L) |
| Normal control group model matched group Rhizoma Anemarrhenae extract group glyburide group | - - 50 100 150 20 | 3.35±0.49 * 13.78±4.67 11.31±9.01 ** 10.64±7.89 ** 9.12±5.98 ** 9.15±5.78 ** |
Annotate: compare with model control group,
*P＜0.01,
*P＜0.05.
Embodiment 7 Rhizoma Anemarrhenae extracts are to the influence of the blood glucose of alloxan diabetes mice
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), (Lik-Sang pharmaceutical factory in Tianjin produces glyburide, lot number 990203), alloxan (production of Sigma company), blood sugar detection test kit (Great Wall, Baoding clinical reagent company, lot number 991514).
2, method
80 of male mice in kunming, fasting 12 hours before the experiment, all the other tail veins inject alloxan 90mg/Kg to 10 of picked at random as the normal control group, 72 hours rear side blood glucose values, choosing wherein 50 blood glucose value＞13m mol/L persons as diabetic mice.Diabetic mice be divided at random high, medium and low three the dosage groups of Rhizoma Anemarrhenae extract (50mg/Kg, 100mg/Kg, 150mg/Kg).Model control group, glyburide positive controls (20mg/Kg) makes there was no significant difference between each blood glucose value mean of organizing, 10 every group.Each organizes gastric infusion or normal saline every day, and continuous 21 days, after the last administration, got blood from mouse orbit in 1 hour, glucose oxidase method is surveyed blood glucose value, the results are shown in Table 4.As shown in Table 4, administration group blood glucose is compared remarkable reduction (P＜0.01) with model control group, and blood glucose descends and Rhizoma Anemarrhenae extract is dose dependent.
Table 4 Rhizoma Anemarrhenae extract is to the blood sugar influence of model induced by alloxan diabetic mice
| Group | Dosage (mg/Kg) | Blood glucose (mmol/L) |
| Normal control group model matched group Rhizoma Anemarrhenae extract group glyburide group | _ _ 50 100 150 20 | 5.29±0.58 23.72±1.51△△ 21.18±1.88 * 19.45±1.63 ** 18.32±1.56 ** 18.94±1.74 ** |
Annotate: compare △ △ P＜0.01 with the normal control group; Compare with model control group,
*P＜0.05,
*P＜0.01.
Embodiment 8 Rhizoma Anemarrhenae extracts are to the influence of the liver glycogen content of alloxan diabetes mice
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), glyburide (Lik-Sang pharmaceutical factory in Tianjin produces, lot number 990203), alloxan (production of Sigma company).
2, method
80 of male mice in kunming, fasting 12 hours before the experiment, all the other tail veins inject alloxan 90mg/Kg to 10 of picked at random as the normal control group, 72 hours rear side blood glucose values, choosing wherein 50 blood glucose value＞13.8m mol/L persons as diabetic mice.Diabetic mice be divided at random high, medium and low three the dosage groups of Rhizoma Anemarrhenae extract (50mg/Kg, 100mg/Kg, 150mg/Kg).Model control group, glyburide positive controls (20mg/Kg) makes there was no significant difference between each blood glucose value mean of organizing, 10 every group.Each organizes gastric infusion or normal saline every day, continuous 21 days, in the last administration after 1 hour disconnected marrow put to death, press sulphuric acid-anthrone method, mensuration liver glycogen content, the result is as shown in table 5:
Table 5 Rhizoma Anemarrhenae extract is to the influence of the liver glycogen content of alloxan diabetes mice
| Group | Dosage (mg/Kg) | Liver glycogen content (mg/g) |
| Normal control group model matched group Rhizoma Anemarrhenae extract group | - - 50 100 150 | 38.21±1.65 18.67±1.47△△ 21.28±2.28 * 23.37±4.18 ** 27.49±4.24 ** |
| The glyburide group | 20 | 26.67±4.75 ** |
Annotate: compare △ △ P＜0.01 with the normal control group; Compare with model control group,
*P＜0.05,
*P＜0.01.
Embodiment 9 Rhizoma Anemarrhenae total extracts are to blood insulin and the morphologic influence of islet tissue of alloxan diabetes mice
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), alloxan (production of Sigma company), anthrone reagent (5-linked chemical plant, Shanghai, lot number 9806).
2, method
60 of male mice in kunming, fasting 12 hours before the experiment, all the other tail veins inject alloxan 90mg/Kg to 10 of picked at random as the normal control group, 72 hours rear side blood glucose values, choosing wherein 30 blood glucose value＞13.8m mol/L persons as diabetic mice.Diabetic mice be divided at random two dosage groups of Rhizoma Anemarrhenae extract height (50mg/Kg, 150mg/Kg) and model control group, 10 every group.Each organizes gastric infusion or normal saline every day, continuous 21 days, in the last administration after 1 hour disconnected marrow put to death, from each group, get 3 mice pancreatic afterbody tissues at random respectively, use Gormori aldehyde-fuchsin and H.E colouring method, the observation islet tissue.Observed result sees Table 5.As seen Rhizoma Anemarrhenae extract is to being had certain repair by the destructive insulin secreting cells of alloxan, and increased the secretory granule of beta Cell of islet.The results are shown in Table 6:
The morphological change of table 6 islet tissue
| Group | The Gormori aldehyde fuchsine stain | H.E dyeing |
| Normal control group model matched group | The secretory granule of β cell are dense and be aubergine, and big or small homogeneous, the β cell degranulation that is evenly distributed are serious, and the kernel color is light, the fraction karyopyknosis | The β cellular morphology is normal, structural integrity, cell is girder shape or strand and arranges pancreatic islet endocrine swelling, arranges sparse, the endochylema minimizing |
[0085]
| Rhizoma Anemarrhenae extract group low dose group Rhizoma Anemarrhenae extract group high dose group | The secretory granule of β cell increase, but granule is sparse, and the secretory granule of arranging inhomogeneous β cell increase, and granule is dense, and kernel is clear and bigger | β cellular morphology structure makes moderate progress than model group, and kernel is bigger and less β cellular morphology structure is approaching normal, and endochylema is even, and no cavity still has the swelling phenomenon |
Embodiment 10 Rhizoma Anemarrhenae extracts are to the active influence of alpha-glucosidase
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), acarbose (Bayer medicines and health protection company limited, lot number 101176), 4-nitrophenols-α-D-pyranglucoside (PNPG, Merck company) alpha-D-glucose glycosides enzyme (Fluka company).
2, method
Rhizoma Anemarrhenae extract and acarbose are dissolved in the DMSO buffer, and dilution is the medicinal liquid of variable concentrations (10-400mg/ml), with 2U/ml glucosidase 10 μ l mixings, in 37 ℃ of incubation 15min, add 10mmol/LPNPG 90 μ l,, under the 410nm wavelength, measure the A value in 37 ℃ of incubation 15min, more than be reflected on 96 orifice plates and finish, the reaction cumulative volume is 200 μ l.Each test sample is done 3 multiple holes simultaneously, averages, and repeats 3 experiments.Acarbose is set blank and negative control simultaneously as the positive control of this law.Calculate the suppression ratio of enzymatic activity: suppression ratio=(A negative control-A test sample)/(A negative control-A blank) * 100%.Logarithm with concentration (mg/ml) is done abscissa, and suppression ratio is done the vertical coordinate mapping, and the result as shown in Figure 1.
Embodiment 11 Rhizoma Anemarrhenae extracts are to the influence of normal mouse blood sugar
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), blood sugar detection test kit (Great Wall, Baoding clinical reagent company, lot number 991518).
2, method
40 of normal mouses, male and female half and half are divided into 4 groups at random: the normal control group, irritate the stomach normal saline; High, medium and low dosage group (50mg/Kg, 100mg/Kg 150mg/Kg) irritate the stomach Rhizoma Anemarrhenae extract respectively, and 1 time/d, 3 weeks of continuous use.Last administration 3h posterior orbit is got blood, and glucose oxidase method is surveyed blood glucose value, and the result is as shown in table 7, and the basic, normal, high dosage group of Rhizoma Anemarrhenae extract all can make blood glucose reduce,
Table 7 Rhizoma Anemarrhenae extract is to the influence of normal mouse blood sugar
| Group | Dosage (mg/Kg) | Blood sugar concentration (mmol/L) |
| Normal control group Rhizoma Anemarrhenae extract group | - 50 100 150 | 9.57±1.75 8.08±3.34 * 7.95±2.37 * 7.14±3.64 ** |
Annotate: compare with the normal control group,
*P＜0.05,
*P＜0.01.
Embodiment 12 Rhizoma Anemarrhenae extracts are to the influence of normal mouse carbohydrate tolerance
1, material
Rhizoma Anemarrhenae extract (pressing embodiment 2 preparations), acarbose (Bayer medicines and health protection company limited, lot number 101176), Kunming mouse (pharmacology teaching and research room of Guangdong Pharmaceutical University provides), blood sugar detection test kit (Great Wall, Baoding clinical reagent company, lot number 991516).
2, method
Get 50 of healthy adult male mices, fasting 2 hours is divided into 5 groups at random: high, normal, basic three dosage group (50mg/Kg of normal control group, acarbose group and Rhizoma Anemarrhenae extract, 100mg/Kg, 150mg/Kg), 10 every group, the blood glucose there was no significant difference that each is organized.Respectively give corresponding dosage once after, irritate stomach with 10kg/kg dosage starch, give starch after 3 hours, eye socket blood sampling side fasting glucose.The result is as shown in table 8:
Table 8 Rhizoma Anemarrhenae extract is to the influence of normal mouse blood sugar
| Group | Dosage (mg/Kg) | Blood glucose (mmol/L) |
| Normal control group Rhizoma Anemarrhenae extract group acarbose group | - 50 100 150 20 | 5.96±1.07 5.70±1.38 4.79±0.84 * 4.95±0.57 ** 4.03±1.42 * |
Annotate: compare with the normal control group,
*P＜0.01,
*P＜0.05.
Claims (1)
1. Rhizoma Anemarrhenae extract is characterized in that being prepared from by following method:
(1) after the Rhizoma Anemarrhenae is pulverized, extract 1～3 time with 6～12 times of volume 10%～90% alcoholic solution, each 1～3h is put cold after-filtration, merging filtrate, concentrated thick paste 1; Filter the back residue and add 6～12 times of volume water extraction 1～3 time, each 1～3h, filtered while hot, merging filtrate, concentrate thick paste 2;
(2) get thick paste 1, with 6～12 times of volume water dissolutioies, last macroporous resin column with 3～5 column volume distilled water eluting, discards eluent; With 4～8 column volumes of 40%～80% alcoholic solution eluting, eluent is concentrated into does not have alcohol, filters, and will precipitate drying and crushing, gets extract 1;
(3) get thick paste 2, add 10～20 times of volume water dissolutioies, trapped fluid is got in ultrafiltration, and concentrate drying gets extract 2; Described ultrafiltration is to adopt the ultrafilter membrane of ultrafilter membrane device and molecular cut off 10000～300000 to carry out ultrafiltration;
(4) the united extraction thing 1,2, promptly get Rhizoma Anemarrhenae extract.
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| CN101805766A (en) * | 2010-04-05 | 2010-08-18 | 苏州瑞蓝博中药技术开发有限公司 | Process for extracting active polysaccharide |
| EP2595633A4 (en) * | 2010-07-19 | 2014-01-22 | Remedy Pharmaceuticals Inc | Methods of intravenous administration of glyburide and other drugs |
| CN102166294B (en) * | 2011-04-08 | 2013-04-17 | 匡海学 | Common Anemarrhena asphodeloides extract with immunosuppressive activity and preparation method and medicinal purposes thereof |
| CN103191289B (en) * | 2013-04-08 | 2015-05-13 | 广州中医药大学 | Synchronous preparation method of four effective parts in medicine pair of common anemarrhena rhizome and amur corktree bark and application thereof |
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