Regulatory Efiect Of XiaoYao Powder On HPA Axis In Impaired Glucose Tolerance Rats And Clinical Smdy Of Relation Between Salivary Cortisol And The Glycolipid Metabolism In Patients With T2DM

Objective1.To study the effect of XiaoYao powder (XYW) on glucolipid metabolism in rats treated with high fat feed and chronic stress.2. To screen the optimal dose of4.0g/kg,2.4g/kg,0.8g/kg,0.4g/kg,0.2g/kg XiaoYao powder on HPA axis and glucolipid metabolism in impaired glucose tolerance rats.3. To study the mechanism of XiaoYao powder on glucolipid metabolism in rats treated with high fat feed and chronic stress.4. To assess the hypothalamic-pituitary-adrenal axis function in patients with type2diabete milutes by study the morning salivary cortisol response to awakening, and the glycometabolism lipid metabolism.Methods1. Rats were fed with high fat feed and treated with one of four stressors concluding hanging, constrainting with gauze, constrainting with PVC barrel and fixing with anchor. Rats were divided into five groups:normal group (normal feed), stress model group (distilled water2ml/L), stress and fluoxetion group (10mg/kg).stress and XYW group(4.0g/kg). stress and St John’s wort (300mg/kg). Recording the food weights everyday,body weights of the rats even week and testing GT (Glucose Tolerance Test) after3,5,7weeks administration. Administrate rats with decapitation in8weeks. CRH,ACTH,CORT,INS were determined by ELISA method, TC, TG,HDL-C were determined by GPO-PAP、COD-CE-PAP and PTA-Mg2+method, respectively. Glycogen was assayed with anthrone colorimetry.2. Rats were fed with high fat feed and treated with one of four stressors concluding hanging, constrainting with gauze,constrainting with PVC barrel and fixing with anchor. Rats were divided into six groups:normal group (normal feed), tress model group(distilled water2ml/L), stress and fluoxetion group (10mg/kg),stress and XYW group (4.0g/kg), stress and XYW group(2.4g/kg). stress and XYW group (0.8g/kg). Recording the food weights every day, body weights of the rats every week and testing GT (Glucose Tolerance Test) after1,3,5, weeks administration. Administrate rats with decapitation in6weeks. TC, TG, HDL-C, LDL-C and glycogen were determined.3. Rats were divided into six groups:normal group (normal feed), stress model group (distilled water2ml/L),stress and fluoxetion group (10mg/kg), stress and XYW group(0.8g/kg), stress and XYW group(0.4g/kg), stress and XYW group(0.2g/kg). Other steps are same as step2.4. To collect the trunk blood of rats administrated with five kinds of doses, such as4.0g/kg,2.4g/kg,0.8g/kg,0.4g/kg,0.2g/kg, CRH, ACTH, CORT were determined by ELISA, GRmRNA was tested by RT-PCR. To investigate the effect of XiaoYao powder on HPA axis by gene regulation.5.119cases of type2diabetes patients were collected according to the internalizing standardization. The blood samples were collected, and the serum FBS, FINS, C-P, HbAlc and plasma lipid level including TC, TG, HDL-C, LDL-C, apoA1,apoB, apoAl/B. salivary cortisol samples were collected immediately at awakening and15min there after. The15min salivary cortisol level minus the salivary eortisol at awakening calculated the morning cortisol response. All of these data were computed with statistics. To assess the hypothalamic-pituitary-adrenal axis function in type2diabete patients.ResultsFundamental research1. Compared with weight loss of model group, xiaoyao powder, st John’s wort and fluoxetine can improve it and xiaoyao powder’s effect was better. In oral glucose test, plasma glucose level of xiaoyao powder group at the time of30min was decreased12%, at fourth week, compared with model group, and was decreased8%,20%, respectively, at sixth and eighth week. St John’s wort and fluoxetine groups also had the effect but not better than xiaoyao powder. Meanwhile, xiaoyao powder group could improve spleen index,abnormal blood lipid level and St John’s wort group could improve liver index.2. Three dose of xiaoyao powder such as4.0g/kg、2.4g/kg、0.8g/kg all had effect on abnormal glucose tolerance at the time of30min and the effect had negative correlation with dose.0.8g/kg was the optimum dose. Meanwhile,0.8g/kg of xiaoyao powder group could improve spleen index, liver index, body weight and food weight. Xiaoyao powder’s dose was positive related with the effect on abnormal blood lipid and insulin.3. Three dose of xiaoyao powder such as0.8g/kg、0.4g/kg、0.2g/kg all had effect on abnormal glucose tolerance at the time of30min but it had not dose-effect relationship.0.8g/kg was the optimum dose and it could improve spleen index, liver index, body weight, food weight, abnormal blood lipid and insulin 4. Preliminary pharmacodynamics experiment showed that GR mRNA level was increased40%,30%,55%respectively by three doses of xiaoyao powder such as4.0g/kg、2.4g/kg、0.8g/kg. Meanwhile, CRH was decreased16%,7%,2%respectively by three doses; ACTH was decreased27%,20%,12%respectively; CORT was decreased14%by4.0g/kg. Pharmacodynamic test showed that GR mRNA level was increased70%,55%,60%respectively by three doses of xiaoyao powder such as0.8g/kg、0.4g/kg、0.2g/kg. Meanwhile, CRH was decreased6%,5%.4%respectively by three doses; ACTH was decreased14%,7%,14%respectively; CORT was decreased10%,10%,6%respectively. Clinical researchMorning salivary cortisol levels in T2DM patients were positively correlated with HbAlc levels (P<0.01). The better the blood glucose be controlled,the higher morning salivary cortisol levels which stand for function of HPA axis. HPA axis sensitivity was negative related with BM1and WRI, and not related with insulin. HPA axis sensitivity was related with lipid metabolism, but the correlation coefficient was not high.Conclusion1. Model group have lower food weights, body weights, hepatic glycogen, abnormal blood lipid level and GT. all of this were improved by XYW.2.0.8g/kg is the optimal dose.3. XiaoYao powder ameliorates the expression of GR by inhibiting the expression of GR mRNA, so that HPA axis accentuation is improved,influencing CRH level.4. There have associatively between the morning salivary cortisol response to awakening, and the glycometabolism lipid metabolism.