Studies On The Insulin Resistance And Glucose Transporter Protein Expression Of Type 2 Diabetes By Wenyang Huoxue Recipe

Purpose:In traditional chinese medicine,diabetes features excessive discharge of urine and persistent thirst improper,which is caused by improper diet, emotional disorders and weakness.Insulin resistance is the important cause of the occurrence and development of type 2 diabetes.Traditional Chinese medicine believes that the basic pathogenesis of diabetes is Yang deficiency and blood stasis.Spleen kidney yang deficiency is the initiating factor in diabetes.Blood stasis is the basic pathological changes of diabetes and persists in the whole diseases process,with is the reasons for deterioration and complications of diabetes.This theory attributes the initial and deteriorating factors of diabetes to organ dysfunction,which conforms to the modern view of diabetes:a disease of ineffective energy use of body.The research and treatment targeted Yang deficiency and blood stasis will help to prevent many complications and truly improve the quality of life in diabetes patients.Moreover,through studies on the constitution of vulnerable groups, it will do benefit to early prevention of diabetes.We established the Yang deficiency and blood stasis rat model of type 2 diabetes,focusing on the molecules related to Yang deficiency and blood stasis.Identification of these molecules can provide a basis for screening effective components from Chinese herbal medicine.Methods:We established the wistar rat model of type 2 diabetes by application of streptozotocin and high-fat diet.We choose rats with fasting blood glucose (FBG) more than 11.lnmol/L,more drink,more food,overweight,high blood lipids,elevated blood insulin levels and declined insulin sensitivity as successful model of insulin resistance and diabetes.We selected 60 rats for modeling from 70 rats,the left remained as normal controls.The successful models were randomly divided into 6 groups:Group A:normal control;Group B:diabetes model control;Group C:low-dose Wenyang Huoxue recipe,0.101g/kg; Group D:moderate-dose Wenyang Huoxue recipe,0.203g/kg;Group E:high-dose Wenyang Huoxue recipe,0.406g/kg;Group F:Metformin,0.1g/kg.During the experiment each rats was fed water freely.In addition to the normal control group,the remaining rats in each group were injected subcutaneously insulin 1.5IU/d to enhance the survival of animals.Eight weeks after delivery,urine and blood were collected from rats in each group and serum were separated. After rats were killed,quadriceps tissues were separated for cryopreservation. Urinary albumin,urinaryβ2-MG,plasma cAMP and cGMP content and serum insulin were determined by radioimrnunoassay.Blood glucose were determined by glucose oxidase method.RT-PCR and western blot were used to detect the GLUT4 gene expression of skeletal muscle in mRNA and protein levels,respectively.Serum TNF-αand adiponectin were determined by Enzyme-linked immunosorbent assay.Results:Wenyang Huoxue recipe significantly reduced blood glucose levels in diabetic rats,but didn't increase serum insulin.It significantly improved insulin sensitivity index and lowered insulin resistance index.Diabetic rats showed albumin andβ2-MG in urine,which is an indication of early diabetic nephropathy.Wenyang Huoxue recipe showed the protective effect for kidney by lowering urinary albumin andβ2-MG.Diabetes models had lower plasma cAMP and cAMP/cGMP than normal controls,indicating the existence of Yang deficiency.Wenyang Huoxue recipe could increase these two indicators and improve the body's basal metabolic rate.In skeletal muscle cells,rats treated by Onyang Huoxue recipe had higher mRNA and protein expression of GLUT4,with is a molecule promoting uptake and usage of glucose in skeletal muscles. Diabetic rats treated by Onyang Huoxue recipe also had lower TNF-αand higher adiponectin in plasma,indicating the antagonistic role of insulin resistance and micro-inflammatory response.Micro-inflammation can lead to high blood viscosity,endothelial dysfunction and other symptoms of blood stasis in diabetic patients.Conclusions:Huoxue Onyang recipe can reverse insulin resistance and effectively lower blood glucose level in diabetic rats,the mechanism may lie in the promotion of glucose uptake in skeletal muscle cells.It can also change the level of fat cytokines to improve blood stasis and protect kidney.The key molecules regulated by Huoxue Onyang recipe provide a clue to find the more effective components of Chinese herbal medicine.