Experimental Study On The Effects And Mechanism Of Chinese And Western Medicine Hypoglycemic Compound

Objective:To explore the optimal blood glucose assessment point in T2 DM mice;Establish a T2 DM composite mouse model with insulin resistance(IR)and partial ?cell damage;Study the efficacy of the Chinese and Western medicine compound which composed of Liuwei Dihuang soup,Yuquan San,and western medicine;Develop the dismantlement research on traditional Chinese compound prescription and evaluate the hypoglycemic effect of the compatibility of traditional Chinese medicine and explore related mechanism.Method:1.T2 DM mice induced by high-fat diet and streptozotocin(STZ)were used as the experimental group and normal C57BL/6 mice as the control group.Continuous fasting blood glucose and continuous random blood glucose were measured in the two groups,and the blood glucose rules of the mice were analyzed.To evaluate the scientificity of 2-hour postprandial blood glucose as the evaluation point of blood glucose intervention in T2 DM mice.By observing 2-hour postprandial blood glucose of T2 DM mice for 7 weeks to verify the stability of this blood glucose assessment point,and it was selected as the evaluation point of the T2 DM mice modeling evaluation and drug intervention in following study.2.T2 DM composite mouse model: Male C57BL/6 mice were fed with high fat diet for 4 weeks,during which each mouse was subcutaneously injected with 1 mg/kg dexamethasone,7 times every other day,and 5 times intraperitoneally injected with 50 mg/kg STZ.The stability of T2 DM composite mice model was evaluated by long-term observation of water and food intake,weight,and 2-hour postprandial blood glucose.By measuring serum insulin,calculating HOMA-IR and HOMA-? values,to evaluate IR and ? cell function of T2 DM mice.3.On the basis of Liuwei Dihuang soup and Yuquan San,supplemented with hypoglycemic agent I(compound I)and hypoglycemic agent II(compound II)respectively.Xiaoke pill was used as the positive drug.Observe the main effect of Chinese and western medicine hypoglycemic compound on T2 DM mice.The observation period of pharmacodynamics was 7 weeks.The body weight,water and food intake,and 2-hour postprandial blood glucose were observed every week.At the end of the experiment,serum insulin,HOMA-IR and HOMA-? were measured.4.Hypoglycemic effect and the mechanism of Chinese medicine compatibility medicine pair: total of 4 groups of prescriptions included(Pueraria & Trichosanthes,Radix Rehmanniae & Fructus corni,Astragalus & Radix Rehmanniae,Pueraria&Astragalus).Metformin was used as the positive control group.The changes of body weight,water intake and 2-hour postprandial blood glucose were observed during 7 weeks of intervention.Serum,pancreas and liver tissues were collected after administration.Parts of pancreatic tissue was fixed and sectioned for HE staining.Serum insulin(INS),glucagon(GC),tumor necrosis factor-?(TNF-?),interleukin-6(IL-6),liver tissue glucose transporter-4(GLUT-4),insulin receptor substrate-1(IRS-1),Phosphatidylinositol 3-kinase(PI3K),Protein Kinase B(AKT/PKB),signal transducer and activator of transcription(STAT5)and pancreatic STAT5 protein were detected by ELISA.Result:1.The continuous fasting blood glucose of mice in T2 DM showed that it was significantly reduced after fasting for more than 4 hours.In contrast,blood glucose changes in T2 DM at 2-3 hours after meal were relatively stable,which could reflect the stability of the model for a long time.2.High fat diet,1 mg/kg dexamethasone combined with low dose STZ for 5 times can induceT2 DM model.Mice in this model had obvious insulin resistance(IR)and partial damage of ? cells.The blood glucose of the model group was significantly higher than control group(P < 0.01).HOMA value suggested that the model group mice had significant IR and ? cell damage.3.The blood glucose of mice in compound?group decreased to 11.1 ± 3.94 mmol/L at the 4th week of administration;The blood glucose of compound ? group decreased to 11.5 ± 4.68mmol/L at the 5th week of administration,close to normal control group(P> 0.05).Compared with the positive drug Xiaoke pill,the compound ?group was better than Xiaoke pill in maintaining the weight of T2 DM mice,improving insulin resistance and ? cell function.The HOMA-IR value of the compound?group(3.87 ± 1.13)and the compound ? group(3.98 ± 1.73)was lower than Xiaoke Pill group(5.17 ± 1.83).The HOMA-? value of the compound ? group(32.4 ± 10.9)and the compound ? group(22.9 ± 4.3)was higher than the Xiaoke pill group(23.5 ± 6.8).In T2 DM model group,the food and water intake was increased,but with the weight decreased significantly,and has a mortality rate of 22% after 4 weeks.The water and food intake of the compound groups were relatively stable during treatment,and the weight increased significantly after 3 weeks of treatment(P<0.05).It is suggested that this Chinese and western medicine compound has significant effects on alleviating the weight loss of T2 DM mice,and Compound ? is significantly better than Xiaoke Wan in improving IR in diabetic mice.4.Chinese medicine compatibility medicine pairpharmacodynamic test result: In second week of Pueraria & Trichosanthes administration,the blood glucose was significantly decreased than model group(P < 0.05).In fourth week,the blood glucose was no different from control group(P > 0.05).In second week of Radix Rehmanniae & Fructus corni administration,the blood glucose was significantly lower than model group(P< 0.05),and the trend of blood glucose change during administration was similar to metformin group.The HOMA-IR value of Pueraria & Trichosanthes group(5.47± 0.99),Radix Rehmanniae & Fructus corni group(6.59± 1.07)was significantly lower than metformin group(7.83± 1.21).The HOMA-? value of Pueraria & Trichosanthes group(54.38± 11.31)was comparable to the metformin group(54.18± 7.90),and Radix Rehmanniae & Fructus corni group(45.60± 9.07)was significantly lower than metformin group.At the third week of administration,water and food intake in each group were significantly reduced,compared with the model group(P< 0.05).The body weight of Astragalus & Radix Rehmanniae group was significantly reduced compared with control group(P< 0.05)in the second week,and significantly decreased compared with model group(P< 0.05)in the fifth week.The results showed that Chinese medicine compatibility medicine paircan obviously reduce the blood sugar of T2 DM mice and improve the symptoms of diabetes.5.The results of immune cytokines and insulin signal transduction protein showed that serum TNF-? in T2 DM group was significantly higher than control group(P< 0.05).Serum IL-6 of metformin group,Radix Rehmanniae & Fructus corni,Astragalus & Radix Rehmanniae,Pueraria& Astragalus group were significantly higher than control group(P< 0.05).Liver GLUT-4 in each administration group was significantly higher than control group(P< 0.05).Liver IRS-1 of Astragalus & Radix Rehmanniae group was significantly higher than control group(P < 0.05).Liver PI3 K and AKT protein of model group and each administration group was significantly higher than control group(P< 0.05)?Conclusion:1.This study was the first to observe the changes of blood glucose in T2 DM mice within 12 hours through scientific and reasonable experimental design.It was found that 2 hours postprandial blood glucose stably reflect the hyperglycemia of T2 DM mice for a long time,which can be used as the best observation point for model evaluation and drug intervention.2.This study is the first attempt to use high-fat diet,1 mg / kg of dexamethasone combined with low-dose multiple STZ to induce T2 DM mice compound model.In this T2 DM model,hyperglycemia status of mice can be stable for 4 weeks.It also has the characteristics of IR and partial ? cell damage.This T2 DM model is a mouse model that can reflect the characteristics of T2 DM completely.3.The traditional Chinese and Western compound prescription and its basic Chinese medicine can effectively reduce the blood sugar of T2 DM mice,relieve diabetes related symptoms,and to some extent,correct the IR and improve the ? cell function.4.The main mechanism of Chinese medicine compatibility medicine may be related to the up regulation of GLUT4 and IRS-1 / PI3 K / AKT channel protein in the liver and the alleviation of IR in the liver.5.In this study,it was found that TNF-? and IL-6,as the upstream promoter of insulin signaling pathway and JAK / STAT signaling pathway,may play a role in the proliferation and repair of hepatocytes in T2 DM mice,but their biological functions in the pathogenesis and efficacy of T2 DM still need further study.