CN102895342A - Medicinal composition for preventing or/and treating diabetes mellitus and application of medicinal composition - Google Patents
Medicinal composition for preventing or/and treating diabetes mellitus and application of medicinal composition Download PDFInfo
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Abstract
本发明提供了一种预防或/和治疗糖尿病的药物组合物，它是由原料药葛根、黄连制备而成的制剂。本发明还提供了该药物组合物的用途。本发明药物原料葛根、黄连配伍使用，用于预防或/和治疗2型糖尿病，具有协同增效的作用，将药材中的有效部位葛根总黄酮和黄连总生物碱配比使用，也能达到较佳的协同增效的作用，为临床提供了一种新的用药选择。The invention provides a pharmaceutical composition for preventing and/or treating diabetes, which is a preparation prepared from raw materials Pueraria Radix and Coptis Rhizoma. The invention also provides the application of the pharmaceutical composition. The medicinal raw materials Pueraria lobata and Coptis chinensis are used in combination to prevent or/and treat type 2 diabetes, and have a synergistic effect. The combination of the effective parts of Pueraria lobata total flavonoids and Coptis chinensis total alkaloids in the medicinal materials can also achieve a higher The best synergistic effect provides a new drug option for clinical practice.
Description
Technical field
The present invention relates to a kind of prevention or/and the pharmaceutical composition for the treatment of diabetes belongs to drug world.
Background technology
Diabetes (diabetes mellitus, DM) definition: the metabolism disorder take chronic hyperglycemia as feature that is caused by Different types of etiopathogenises, unusual with the sugar, fat and the protein metabolism that cause because of insulin secretion and/or effect defective.This disease main symptom clinically is " three-many-one-little ", namely polydipsia, polyphagia, polyuria, lose weight.Type 2 diabetes mellitus: take insulin resistant as main, companion's hypoinsulinism is to accompanying insulin resistant (WHO1999 standard) take hypoinsulinism as the master.
Type 2 diabetes mellitus also is adult's morbidity type diabetes, how to fall ill after 35 ~ 40 years old, accounts for diabetics more than 90%.The ability that produces insulin in the type 2 diabetes mellitus ward mate body is not to completely lose, insulin even produce too much in the patient body that has, but the action effect of insulin has a greatly reduced quality, so the insulin in the patient body may be in a kind of state of relative shortage.Can stimulate by some oral drugs the secretion of insulin in the body.But still have the part client need as type 1 diabetes, to carry out insulinize to the later stage.In a word, the short-term goal of type 2 diabetes mellitus treatment is control blood glucose, and long term object is generation and the development of prevention related complication.Its Primary Care scheme mainly is made of motion and diet, but Drug therapy and blood sugar monitoring are often also very crucial.
GEGEN QINLIAN TANG for curing holy Zhang Zhongjing recipe, is recorded in during treatise on Febrile Diseases distinguishes sun abnormal pulse card and control, and is comprised of Radix Puerariae, Radix Scutellariae, Rhizoma Coptidis and Radix Glycyrrhizae four flavor Chinese medicines, has the effect of the cleaning of inducing sweat.Put down in writing in the book: " half jin of Radix Puerariae, two liang in Radix Glycyrrhizae is processed three liang of Radix Scutellariaes, three liang of Rhizoma Coptidis.Upper four flavors with eight liters in water, boil first Radix Puerariae, subtract two liters, receive all medicines, boil and get two liters, go dregs, minute secondary decoction being taken warmly ".Clinical and modern study shows in a large number, GEGEN QINLIAN TANG is widely used in the treatment of diabetes, effect is similar with metformin, especially diabetics is had dry mouth with bitter taste mostly, scorching hot under dysphoria with smothery sensation or the heart in the heart, or the acid regurgitation belch, red tongue with yellow fur, the symptoms such as rapid pulse have preferably therapeutical effect, and are consistent with prescription structure and the effect of GEGEN QINLIAN TANG, and this has also embodied the clinical syndrome differentiation thought of the traditional Chinese medical science " treating different diseases with the same therapeutic principle ".
Summary of the invention
Technical scheme of the present invention has provided a kind of prevention or/and treat the pharmaceutical composition of diabetes, and another technical scheme of the present invention has provided preparation method and the purposes of this pharmaceutical composition.
The invention provides a kind of prevention or/and the pharmaceutical composition for the treatment of diabetes, it is the preparation that the crude drug by the following weight proportioning is prepared from:
Radix Puerariae 1-10 part, Rhizoma Coptidis 1-10 part.
Further preferably, it is the preparation that the crude drug by the following weight proportioning is prepared from:
Radix Puerariae 2-9 part, Rhizoma Coptidis 2-9 part.
Still more preferably, it is the preparation that the crude drug by the following weight proportioning is prepared from:
8 parts of Radix Puerariaes, 3 parts of Rhizoma Coptidis.
Wherein, described Rhizoma Coptidis comprises living product Rhizoma Coptidis and processed product thereof.
Medicine of the present invention is to be active component by the water of Radix Puerariae, Rhizoma Coptidis or extractive with organic solvent, adds the preparation that pharmaceutically acceptable adjuvant or complementary composition are prepared from.
Wherein, described preparation is oral formulations.
A kind of prevention also is provided in the present invention or/and the pharmaceutical composition for the treatment of diabetes, it is to be active component by the effective site Rhizoma Coptidis total alkaloids in effective site Radix Puerariae total flavones, Rhizoma Coptidis and the processed product thereof in the Radix Puerariae, add that pharmaceutically acceptable adjuvant or complementary composition are prepared into preparation pharmaceutically commonly used, wherein, the weight proportion of Radix Puerariae total flavones and Rhizoma Coptidis total alkaloids is:
Radix Puerariae total flavones 1-100 part, Rhizoma Coptidis total alkaloids 1-100 part.
Wherein, the weight proportion of described Radix Puerariae total flavones and Rhizoma Coptidis total alkaloids is:
Radix Puerariae total flavones 1.68-7.56 part, Rhizoma Coptidis total alkaloids 2.513-11.31 part.
Further preferably, the weight proportion of described Radix Puerariae total flavones and Rhizoma Coptidis total alkaloids is:
5.6 parts of Radix Puerariae total flavoness, 2.6385 parts of Rhizoma Coptidis total alkaloidss.
The present invention also provide described pharmaceutical composition in preparation prevention or/and treat diabetes, improve insulin resistant, the purposes in the medicine of blood fat reducing diabetes.
Wherein, described medicine is that prevention is or/and the medicine for the treatment of type 2 diabetes mellitus.
Medicine material Radix Puerariae of the present invention, Rhizoma Coptidis compatibility use, be used for prevention or/and the treatment diabetes, effect with Synergistic, effective site Radix Puerariae total flavones in the medical material and Rhizoma Coptidis total alkaloids proportioning are used, also can reach the effect of better Synergistic, provide a kind of new medication to select for clinical.
The specific embodiment
The preparation of embodiment 1 medicine of the present invention
Get crude drug Radix Puerariae 40g, Rhizoma Coptidis 15g, decoct with water, be prepared into decoction.
The preparation of embodiment 2 medicines of the present invention
Get crude drug Radix Puerariae 1g, Rhizoma Coptidis 10g adds 70% ethanol extraction, and is concentrated, adds starch, and granule processed gets granule.
The preparation of embodiment 3 medicines of the present invention
Get crude drug Radix Puerariae 10g, Rhizoma Coptidis 10g, decoct with water, concentrated, add starch, to granulate, tabletting gets tablet.
The preparation of embodiment 4 medicines of the present invention
Get Radix Puerariae total flavones 100mg, Rhizoma Coptidis total alkaloids 100mg, mixing adds starch, granulates, and tabletting gets tablet.
Wherein, the extraction process of Radix Puerariae flavone constituents, Rhizoma Coptidis alkaloid constituents
Get the Radix Puerariae medical material, adopt 30% ethanol, 7 times of amounts, supersound extraction 30min, supersound extraction once, extracting solution is concentrated into 1:2(medical material weight and medicine liquid volume ratio) for subsequent use, get Radix Puerariae total flavones.General flavone content is about about 42mg/mL in the extracting solution after measured.
Get Rhizoma Coptidis, take water as extracting solvent, use respectively 12,10,10 times of amounts, extract 3 times, soak time is 0.5h, and reflux extracting time is 1.5h, 1.5h, 1h respectively.Decocting liquid is concentrated into 1:10(medical material weight g and medicine liquid volume ml ratio) for subsequent use, get Rhizoma Coptidis total alkaloids.Yield of extract 25.13%, content of berberine hydrochloride are 2.36% with this understanding after measured.
The preparation of embodiment 5 medicines of the present invention
Get Radix Puerariae total flavones 11.2g alkaloid 5.277g, add starch, direct compression gets tablet.
The preparation of embodiment 6 medicines of the present invention
Get Radix Puerariae total flavones 16.8g, Rhizoma Coptidis total alkaloids 113.1g, add starch, granulate, get granule.
The preparation of embodiment 7 medicines of the present invention
Get Radix Puerariae total flavones 7.56g, Rhizoma Coptidis total alkaloids 2.513g, mix, encapsulated, get capsule.
The method for extraction and purification of Radix Puerariae total flavones of the present invention, Rhizoma Coptidis total alkaloids not only is confined to said extracted technique, also can adopt other method preparation of present bibliographical information.
Below prove beneficial effect of the present invention by concrete pharmacodynamics test.
The best compatibility test of test example 1 medicine material Radix Puerariae Rhizoma Coptidis of the present invention
1, the design of proportioning
Based on baseline geometric ratio increase and decrease method, press Radix Puerariae Rhizoma Coptidis total amount 55g and calculate, design respectively Radix Puerariae and Rhizoma Coptidis following proportioning: 11:0,9:2,8:3,6:5,4:7,2:9,0:11 totally 7 groups carry out effectiveness and test,
Preparation method: take by weighing each single medicine in said medicine composition and ratio, soak 20min, according to the method for " Radix Puerariae that is decocted first, then receive all medicines " in the treatise on Febrile Diseases, Radix Puerariae 30min is decocted first, add again other medical materials and decoct 30min, filter, decoct again 30min, filter, merge medicinal liquid, concentrated ,-20 ℃ save backup.
With the centrifugal 10min of each drug extract 8000r/min, draw supernatant before the experiment, 0.22 μ m microporous filter membrane positive press filtration degerming, packing, 4 ℃ save backup.
Cell experiment calculates by above-mentioned effective dose 82.5mg crude drug/L, is grouped as follows, and sees Table 1.
Table 13T3-L1 test cell line treated in vitro dosage
2, based on the Radix Puerariae Rhizoma Coptidis proportioning test of adipose cell before the 3T3-L1
2.1, before the 3T3-L1 adipose cell cultivation and induce differentiation
Adipose cell suspension before the 3T3-L1 is pressed 5 * 10
5The density of/ml is inoculated into 48 well culture plates, in the DMEM high glucose medium that contains 10% calf serum, at 37 ℃, 5%CO
2Cultivate under the saturated humidity condition, 2d changes liquid 1 time.After cell fusion 2d reaches contact inhibition, DMEM complete culture solution with the Dex that contains 1 μ mol/L, 0.5mmol/LIBMX and 5mg/L insulin is induced differentiation, remove Dex and IBMX behind the 48h, with 10mg/L insulin continuation effect 48h again, change normal complete culture solution and cultivate 8 ~ 12d, change every other day liquid, until 3T3-L1 cell more than 90% is used for test when being the adipose cell phenotype.2.2 medicine is on the impact of 3T3-L1 adipose cell insulin resistant (IR) model glucose utilization
After cell broke up fully, except blank group adds normal culture medium, all the other each groups all added the dexamethasone of 1 μ mol/L, the insulin of 1 μ mol/L carries out the insulin resistant of adipose cell, effect 3d.After the adipose cell insulin resistant is set up, use instead without phenol red DMEM high glucose medium, add respectively the tested pharmaceutical intervention 3d of variable concentrations, medicine is observed medicine to the improvement effect of insulin resistant with table 1.Get the content of cell culture supernatant 505nm colorimetric determination glucose behind the 72h, calculate grape cell sugar and utilize the relative change rate.
Glucose utilization rate of change (%)=(1 ﹣ experimental group/model group) * 100%
Adipose cell is adipose cell and identifies before inducing respectively 3T3-L1.Adopt the dexamethasone of 1 μ mol/L, the insulin-induced adipose cell of 1 μ mol/L to set up insulin resistant model.Add respectively the tested pharmaceutical intervention 3d of variable concentrations by table 1, observe medicine to the improvement effect of insulin resistant.Get the content of cell culture supernatant 505nm colorimetric determination glucose behind the 72h, calculate grape cell sugar and utilize the relative change rate.
Glucose utilization rate of change (%)=(1 ﹣ experimental group/model group) * 100%
The different proportionings of table 2 Radix Puerariae Rhizoma Coptidis are to 3T3-L1 adipose cell IR model
The impact of glucose utilization
Annotate: compare with model group,
*P＜0.05,
*Same under P＜0.01(); The dosage unit of rosiglitazone is μ mol/L.
By table 2 result as can be known, compare with the blank group, the 3T3-L1 adipose cell is under the dexamethasone and insulin effect, and model group obviously reduces the sensitivity of insulin, the picked-up of glucose obviously reduces in the culture fluid, shows adipose cell insulin resistant model modeling success.Compare with model control group, the different proportionings of Radix Puerariae Rhizoma Coptidis all can be obviously or part reduce the content (all P＜0.05) of glucose in the culture fluid, improve the utilization rate of adipose cell glucose, improve insulin resistant, act on similar to the positive drug rosiglitazone, especially be better than single Radix Puerariae and single Rhizoma Coptidis with two medicine compatibility curative effects, take the Radix Puerariae Rhizoma Coptidis by the proportioning of 8:3 as optimal dose, utilization rate reaches 24.0% relatively.
Test example 2 medicine material compatibility preliminary study tests of the present invention
The setting of this research dosage, the Eastern Han Dynasty of again investigating according to nineteen eighty-three professor Ke Xuefan and the conversion relation of modern medicines dosage, in conjunction with Chinese department of Chinese medicine institute with holt [Zhao Linhua, Lian Fengmei, Ji Hangyu, Deng. professor Tong Xiaolin uses various dose GEGEN QINLIAN TANG treatment type 2 diabetes mellitus to test case. Chinese experimental pharmacology of Chinese medical formulae magazine .2011,17 (4): 249.] and Liu An [Liu Yuzheng, chapter army. Wang Yuesheng, Deng. GEGEN QINLIAN TANG dosage Discussions of The Related Issues. Chinese experimental pharmacology of Chinese medical formulae magazine .2011,16 (16): 216.] etc. to the discussion of GEGEN QINLIAN TANG dosage relevant issues, and " Chinese pharmacopoeia and the party's modern study, determine that the ratio between the Radix Puerariae Radix Astragali Rhizoma Coptidis Radix Glycyrrhizae is 8:3:3:2 in the GEGEN QINLIAN TANG, the Coming-of-Age Day of GEGEN QINLIAN TANG takes dosage and is: Radix Puerariae 40g, Radix Scutellariae 15g, Rhizoma Coptidis 15g, Radix Glycyrrhizae 10g.So it is that 80g/d(becomes body weight for humans to calculate by 60kg that the Coming-of-Age Day of GEGEN QINLIAN TANG takes dosage), i.e. 1.3g/kg.
The 1 GEGEN QINLIAN TANG side of tearing open the research based on adipose cell before the 3T3-L1
1.1 the preparation of medicine and grouping
Take by weighing each single medicine in said medicine composition and ratio, soak 20min, according to the method for " Radix Puerariae that is decocted first, then receive all medicines " in the treatise on Febrile Diseases, Radix Puerariae 30min is decocted first, add again other medical materials and decoct 30min, filter, decoct again 30min, filter, merge medicinal liquid, concentrated ,-20 ℃ save backup.
With the centrifugal 10min of each drug extract 8000r/min, draw supernatant before the experiment, 0.22 μ m microporous filter membrane positive press filtration degerming, packing, 4 ℃ save backup.According to the cell viability preliminary experiment result of GEGEN QINLIAN TANG perfect square in early stage, grouping and dosage are as follows.
Table 33T3-L1 test cell line treated in vitro dosage
Annotate: each group is carried out compatibility by Radix Puerariae 40g, Radix Scutellariae 15g, Rhizoma Coptidis 15g, Radix Glycyrrhizae 10g, adopts said method to be prepared into decoction and carries out the test of pesticide effectiveness.
1.23T3-L1 the cultivation of front adipose cell and induce differentiation
Adipose cell suspension before the 3T3-L1 is pressed 5 * 10
5The density of/ml is inoculated into 48 well culture plates, in the DMEM high glucose medium that contains 10% calf serum, at 37 ℃, 5%CO
2Cultivate under the saturated humidity condition, 2d changes liquid 1 time.After cell fusion 2d reaches contact inhibition, DMEM complete culture solution with the Dex that contains 1 μ mol/L, 0.5mmol/LIBMX and 5mg/L insulin is induced differentiation, remove Dex and IBMX behind the 48h, with 10mg/L insulin continuation effect 48h again, change normal complete culture solution and cultivate 8 ~ 12d, change every other day liquid, until 3T3-L1 cell more than 90% is used for test when being the adipose cell phenotype.
1.3 medicine is on the impact of 3T3-L1 adipose cell insulin resistant (IR) model glucose utilization
After cell broke up fully, except blank group adds normal culture medium, all the other each groups all added the dexamethasone of 1 μ mol/L, the insulin of 1 μ mol/L carries out the insulin resistant of adipose cell, effect 3d.After the adipose cell insulin resistant is set up, use instead without phenol red DMEM high glucose medium, add respectively the tested pharmaceutical intervention 3d of variable concentrations, medicine is observed medicine to the improvement effect of insulin resistant with table 1.Get the content of cell culture supernatant 505nm colorimetric determination glucose behind the 72h, calculate grape cell sugar and utilize the relative change rate.
Glucose utilization rate of change (%)=(1 ﹣ experimental group/model group) * 100%
Table 4 GEGEN QINLIAN TANG and the side of tearing open thereof are to 3T3-L1 adipose cell IR model
The impact of glucose utilization
Annotate: compare with model group,
*P＜0.05,
*Same under P＜0.01(); The dosage unit of rosiglitazone is μ mol/L.
By table 5 result as can be known, compare with the blank group, the 3T3-L1 adipose cell is under the dexamethasone and insulin effect, and model group obviously reduces the sensitivity of insulin, the picked-up of glucose obviously reduces in the culture fluid, shows adipose cell insulin resistant model modeling success.Compare with model control group, Radix Puerariae and Rhizoma Coptidis mutual reinforcement between compatibility, the content that full side more can obviously reduce glucose in the culture fluid improves the glucose utilization rate, improves insulin resistant, and its relative utilization rate reaches 31.09%; Simultaneously, Radix Scutellariae and Radix Glycyrrhizae have to a certain degree antagonism to the blood sugar reducing function of Rhizoma Coptidis, and this effect may be relevant with the change of generation precipitation or extracting solution pH in the decoction process, and on not obviously impact of Radix Puerariae; In addition, monomer medicine Radix Puerariae, Rhizoma Coptidis can obviously reduce the content (all P＜0.05～0.01) of glucose in the culture fluid, but effect is weaker than 2 compatibilities or full side, and Radix Glycyrrhizae shows certain minimizing glucose utilization rate effect, with each drug action reversal.The above results prompting, Radix Puerariae, Rhizoma Coptidis compatibility more are better than or are equal to full side aspect the insulin resistant improving.
Test example 3 Radix Puerariae Rhizoma Coptidis compatibilities of the present invention are on the impact (demonstration test) of 3T3-L1 adipose cell IR model
According to the experimental result of test example 1 and 2, take the effective dose of Radix Puerariae and Rhizoma Coptidis compatibility as middle dosage, adopt 5 times of dilution methods that medicinal liquid is carried out the gradient preparation, see Table 6.Experimental technique is observed the Radix Puerariae Rhizoma Coptidis extract of variable concentrations to the impact of 3T3-L1 adipose cell insulin resistant model with 1.3.
Table 6 Radix Puerariae Rhizoma Coptidis compatibility is on the impact of 3T3-L1 adipose cell IR model glucose utilization
Annotate: compare with model group,
*P＜0.05,
*Same under P＜0.01(); The dosage unit of rosiglitazone is μ mol/L.
By table 6 result as can be known, compare with the blank group, the 3T3-L1 adipose cell is under the dexamethasone and insulin effect, and model group obviously reduces the sensitivity of insulin, the picked-up of glucose obviously reduces in the culture fluid, shows adipose cell insulin resistant model modeling success.Compare with model control group, Radix Puerariae Rhizoma Coptidis compatibility can be obviously or part reduce the content (all P＜0.05) of glucose in the culture fluid, improve the utilization rate of adipose cell glucose, improve insulin resistant, act on similar to the positive drug rosiglitazone, wherein especially act on the most obviously under the concentration of 412.5mg/L with the Radix Puerariae Rhizoma Coptidis, relatively utilization rate reaches 32.77%.
In sum, Radix Puerariae Rhizoma Coptidis compatibility (8:3) in that to improve aspect the insulin resistant (IR) effect more obvious, and is better than each single drug use than the full side of GEGEN QINLIAN TANG.
The drug efficacy study of test example 4 Radix Puerariae Rhizoma Coptidis compatibility control type 2 diabetes mellitus of the present invention
According to above-mentioned cell and best compatibility proportioning test result of study, this research is intended further observing Radix Puerariae, Rhizoma Coptidis and processed product thereof to the impact of type 2 diabetes mellitus mouse model and euglycemia thereof, determines the effect of compatibility of drugs from the angle of whole animal pharmacodynamics.
1, medicine is on the impact of normal mouse blood sugar
60 of ICR mices are divided into 8 groups by the weight stratified random during experiment, press table 7 design grouping and dosage gastric infusion, every day 1 time, continuously 7d.Water 8h is can't help in the 45min(fasting after the last administration), Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured mice fasting glucose (FBG).
Table 7 is on the impact of mice fasting glucose
Compare P with normal group〉0.05.
By table 7 result as can be known, compare with the blank group, no matter Radix Puerariae is with to give birth to product Rhizoma Coptidis or Chinese goldthread processed product (processed with wine, vinegar system) compatible, all normal mouse fasting glucose (FBG) obviously do not affected (P〉0.05), but respectively organizes the trend that high dose all has blood sugar lowering.
.2 medicine is on the impact of alloxan diabetes mouse model
The ICR mice, male and female half and half, weight 18～22g.Be divided into 2 groups by the weight stratified random during experiment, except 10 be the blank group, all the other mices are the model discrimination group, are used for the screening of mice zoic model with hyperglycemia.Mice fasting 18h before the test.Next day, the equal tail vein injection of all the other mices (iv) alloxan solution 60mg/kg caused mice hyperglycemia model (normal saline of the capacity such as blank group injection) except the blank group, and 4h gavage 50% glucose solution after the injection, 0.4ml/ are only.Behind the modeling 72h, Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured mice fasting glucose (FBG), selects the qualified mice of blood glucose to include formal test in, and the model success standard of fasting glucose is 〉=11.0mmol/L.
Select above-mentioned one-tenth mould mice and blank group mice, be divided into 9 groups by blood glucose and weight stratified random, every group 10, i.e. blank group, model control group, metformin hydrochloride matched group, Ge Lian (life) high and low dose group, Ge Lian (processed with wine) high and low dose group and Ge Lian (vinegar system) high and low dose group.Each organizes mice by table 8 design dosage gastric infusion, every day 1 time, continuously 14d.40min after the administration of 14d last, Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured the fasting glucose (FBG) of mice; Simultaneously, mouse orbit is got blood, the EDTA anticoagulant, and the centrifugal 10min of 800r/min abandons supernatant, after the normal saline washing precipitation, the preparation packed red cells.Add cold distilled water in the long-pending erythrocyte of the ratio pressure of 2:3 and prepare hemolysate, press the content that the kit measurement description is measured glycolated hemoglobin (GHb).
Table 8 is on the impact of mice FBG and GHb
Annotate: compare with model group,
*P＜0.05,
*P＜0.01.
As shown in Table 8, compare with the blank group, behind the model control group mouse tail vein injection alloxan 72h, the symptoms such as polyphagia, polydipsia, polyuria all appear, the content of model control group mice fasting glucose (FBG) and glycolated hemoglobin (GHb) obviously raises, and shows that alloxan causes diabetes mice model modeling success.Compare with model control group, no matter Radix Puerariae and the product of giving birth to Rhizoma Coptidis or compatible with Chinese goldthread processed product (processed with wine, vinegar system), all can be obviously or part reduce the content of mice fasting glucose (FBG) and glycolated hemoglobin (GHb), act on and almost being equal to.
In sum, the inside and outside model of coalition, the treatment " quenching one's thirst " sick aspect, the compatible use of Radix Puerariae and Rhizoma Coptidis and processed product thereof, curative effect is clear and definite, has the glycometabolic effect of clear and definite improvement.
The effective substance development test that test example 5 medicine material Radix Puerariaes of the present invention and Rhizoma Coptidis " are only quenched one's thirst "
Theoretical according to the herbal medicine efficacy component, seminar further carries out compatible combination with the active component isoflavone position of Radix Puerariae and the active component alkaloid position of Rhizoma Coptidis, observes the Different therapeutical effect of component compatibility and crude drug compatibility.
1 material
1.1 medical material and reagent
Determining and grouping of the dosage of raw material
For ease of comparing the Different therapeutical effect of Radix Puerariae Rhizoma Coptidis component compatibility and crude drug compatibility, the clinical every daily dose of the adult of crude drug is Radix Puerariae 40g, Rhizoma Coptidis 15g, simultaneously, the dosage of Radix Puerariae total flavones take crude drug still as 40g, the dosage of Rhizoma Coptidis total alkaloids is primary crude drug 15g, Radix Puerariae Rhizoma Coptidis ratio (by crude drug) is 8:3, is grouped as follows:
Table 9 mouse experiment dosage
Annotate: wherein Ge Lian component Raw Radix Puerariae, Rhizoma Coptidis weight proportion are 8:3, press the method for embodiment 4 and extract Radix Puerariae total flavones and Rhizoma Coptidis total alkaloids.
Metformin hydrochloride tablet, the Zhonghui Pharmaceutical Co.,Ltd., Beijing produces, the accurate word of traditional Chinese medicines: H19983069, lot number: 20110415.All prepare with distilled water during the said medicine experiment.Alloxan, streptozotocin (STZ) are all available from Sigma company.
1.2 laboratory animal ICR mice, cleaning level, male and female half and half, weight 18～22g is produced by Sichuan Academy of Medical Sciences institute of lab animals, the animal quality certification number: the SCXK(river) 2008-24.
1.3 doubly easy type (the One Touch UltraEasy) blood glucose meter of the steady person of outstanding talent of experimental apparatus, Johnson ﹠ Johnson (China) medical apparatus and instruments company limited; Thermo Multiskan MK3 type microplate reader, Thermo Formo company.
2 methods
2.1 on normal fasting glucose (FBG) affect 60 of ICR mices, be divided into 6 groups by the weight stratified random during experiment, press table 9 design grouping and dosage gastric infusion, every day 1 time, continuously 7d.Water 8h is can't help in the 45min(fasting after the last administration), Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured mice fasting glucose (FBG).
2.2 the alloxan diabetes mouse model affected ICR mice, male and female half and half, weight 18～22g.Be divided into 2 groups by the weight stratified random during experiment, except 10 be that all the other mices are the model discrimination group the blank group, be used for the screening of mice hyperglycemia model.Mice fasting 18h before the test.The equal tail vein injection of mice in morning next day (iv) alloxan solution 60mg/kg causes mice hyperglycemia model (normal saline of the capacity such as blank group injection).Behind the modeling 72h, Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured mice fasting glucose (FBG), and qualified mice is included formal test in to select blood glucose, and the model success standard of fasting glucose is 〉=11.0mmol/L.Select the qualified mice of above-mentioned one-tenth mould, be divided into 7 groups by blood glucose and weight stratified random, press table 9 design grouping and dosage gastric infusion, every day 1 time, continuously 14d.40min after the administration of 14d last, Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured the fasting glucose (FBG) of mice; Simultaneously, mouse orbit is got blood, and the EDTA anticoagulant is surveyed respectively the content of insulin (Ins) and glycolated hemoglobin (GHb) by the test kit description, and calculates insulin sensitivity index (IAI).
2.3 the impact on the compound type 2 diabetes mellitus mouse model of the high fat of STZ
The ICR mice, male and female half and half, weight 18～22g.Except modeling changes the blank group injection of mouse peritoneal injection STZ buffer 100mg/kg(equivalent 0.1mol/L citric acid-sodium citrate buffer into), all the other model discrimination methods are same 1.5.2 all.The qualified mice of above-mentioned one-tenth mould is divided into 7 groups by blood glucose and weight stratified random, presses table 9 design grouping and dosage gastric infusion, and every day 1 time, 14d gives the high sugared high salt emulsion 0.4ml/ of high fat and only copies the type 2 diabetes mellitus mouse model in administration continuously.40min after the administration of 14d last, Mouse Tail-tip is got blood, and One Touch UltraEasy type blood glucose meter is measured the fasting glucose (FBG) of mice; Mouse orbit is got blood simultaneously, and part blood EDTA anticoagulant is surveyed respectively the content of insulin (Ins) and glycolated hemoglobin (GHb) by the test kit description, and calculates insulin sensitivity index (IAI); The centrifugal 10min of another part blood 3000r/min, separation of serum is measured respectively the content of mice serum cholesterol (TC) and triglyceride (TG) by the kit measurement description.
3 results
3.1 the impact on normal fasting glucose (FBG)
Table 10 is on the impact of normal mouse FBG
Compare P＜0.05 with normal group.
By table 10 result as can be known, compare with the blank group, can obviously reduce fasting glucose (FBG) content (P＜0.05) of normal mouse behind Radix Puerariae isoflavone and the wine steaming Rhizoma Coptidis alkaloid compatibility.
3.2 the impact on the alloxan diabetes mouse model
Table 11 is on the impact of alloxan mice FBG and GHb
Annotate: compare with model group, * P＜0.05, same under * * P＜0.01().
Table 12 is on the impact of alloxan mice Ins and IAI
By table 11～12 as can be known, compare with the blank group, behind the model group mouse tail vein injection alloxan 72h, the symptoms such as polyphagia, polydipsia, polyuria all appear, the content of model group mice fasting glucose (FBG) and glycolated hemoglobin (GHb) obviously raises, plasma insulin (Ins) content and insulin sensitivity index (IAI) obviously reduce, and show that alloxan causes diabetes mice model modeling success.Compare with model group, all can be obviously behind Radix Puerariae Rhizoma Coptidis and the Radix Puerariae Rhizoma Coptidis component compatibility or part reduce the content of mice FBG and GHb, the content of rising Ins improves the sensitivity (P＜0.05～0.01) of IAI, especially more excellent with Radix Puerariae component blood sugar lowering effect, the blood sugar lowering rate reaches 36.12%.
3.3 the impact on the compound type 2 diabetes mellitus mouse model of the high fat of STZ
Table 13 is on the impact of STZ diabetic mice FBG and GHb
Table 14 is on the impact of STZ diabetic mice Ins and IAI
Table 15 is on the impact of STZ diabetic mice TC and TG
By table 13～15 as can be known, compare with the blank group, the content of model group mouse blood sugar FBG, GHb obviously raises, blood plasma Ins and IAI obviously reduce, the content of blood fat TC, TG also obviously raises, the disorder of glycolipid metabolism appears in model mice, shows that STZ causes the compound type 2 diabetes mellitus model of mice modeling success.Compare with model group, Radix Puerariae Rhizoma Coptidis and Radix Puerariae Rhizoma Coptidis component compatibility all can be obviously or part reduce the content of mice FBG, GHb, TC and TG, the content of rising Ins, improve the sensitivity (P＜0.05～0.01) of IAI, it is disorderly to demonstrate certain adjusting glycolipid metabolism, improves the effect of insulin resistant.
The result of consolidated statement 10～15 as can be known, compare with the blank group, the content of model control group mouse blood sugar FBG, GHb obviously raises, and the content of blood fat TC, TG obviously raises, the disorder of glycolipid metabolism appears in model mice, shows that STZ causes the compound type 2 diabetes mellitus model of mice modeling success.Compare with model control group, Radix Puerariae Rhizoma Coptidis and Radix Puerariae Rhizoma Coptidis component compatibility all can be obviously or part reduce the content of mice FBG, GHb, TC and TG, demonstrate the effect of certain adjusting glycolipid metabolism disorder, especially aspect blood fat reducing, Radix Puerariae Rhizoma Coptidis component curative effect is more excellent.
In sum, Radix Puerariae total flavones and Rhizoma Coptidis total alkaloids compatibility have the obvious glycolipid metabolism disorder that improves, and improve the effect of insulin resistant, and the blood fat reducing drug effect is better than the former medicine compatibility of Radix Puerariae Rhizoma Coptidis.
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