| Objective:In this study, our aim is to observe the effect of Dan-gua prescription on the blood sugar blood lipids and anti-oxidative capacity of genetically induced diabetic mice by the level of ROS, GSH, SOD, SRF and other indicators observed in diabetic ratS. To investigate the mechanism of Dan-gua prescription against oxidative stress, thus to provide a theoretical basis for the clinical guide of diabetes medicine medication.Methods:l.Take8-week-old male mice, weighing at (20±0.5) g of C57BL/6J as normal control group, fed with normal diet, without any treatment. Then we picked35ApoE-/-mice with the same age and weight, continueously injected with STZ40mg/(kg.d) for3days to induce diabetic rats model. Select32mice from the successful induced models, randomly divided them into model group, Dan-gua prescription group, the combined treatment group (Dan-gua prescription+pioglitazone) and pioglitazone group,8in each group. Each group were given medicine respectively, and fed with normal diet for12weeks. Monitoring blood glucose, body weight, skin temperature, once every two weeks.2.Take blood from tail vein to test fasting glucose (FPG), glycosylated hemoglobin (HbAlc), and lipids,after12weeks.3.Take the liver and heart homogenates production to measure related indicators. The Coronary was observed under light microscopy.The content of ROS in liver homogenates was detected by Chemical fluorescence methed, The content of GSH、SOD in liver homogenates were detected by Colorimetry.The expression of SRF in heart homogenates was detected by immunohistochemical.4.All the experimental data were analyzed by using SPSS18.0software. Difference comparison of measurement data among groups were determined with single factor variance analysis.Results:1. Blood glucose and lipid(1) Fasting blood glucose:Four weeks after intervention,the blood sugar of ApoE-/-mice groups were slightly lower than before. While the blood sugar of the model group rised gradually. Dan-gua prescription group, the combined treatment group and pioglitazone group had large fluctuations in blood sugar level from four weeks to eight weeks.Dan-gua prescription group and the combined group of blood glucose values have not changed much after the first eight weeks,while pioglitazone group blood glucose levels tend to rise.Compared with normal control group, There is no significant difference in both of the Dan-gua prescription group and the combination group, but both are significantly lower than that in the model group in12weeks (P<0.01); and the blood sugar level in the pioglitazone group was higher than that in normal control group (P<0.01).(2) Hemoglobin:Model group was significantly higher than the normal control group (P <0.01), there is no significant difference among the treatment groups,but they were significantly lower than that in the model group (P<0.01).(3) Blood lipids:The level of LDL、TC in Dan-gua prescription group were significantly lower than that of model group (P<0.05).2. Oxidative factors:(1) ROS activity was measured in liver homogenates:The level of ROS of the Dan-gua prescription group, pioglitazone group and combined group was significantly lower than that in model group(P<0.05, P<0.05, P<0.05). Compared with normal control group, the pioglitazone group and model group were significantly differences (P<0.05, P=0.014).There is no significant difference between the Dan-gua prescription group and combination group (P=0.798)(2) SOD activity was measured in liver homogenates:Compared with normal control group, the model group and pioglitazone group were significantly differences (P<0.05, P=0.002) The SOD of Dan-gua prescription group, combined treatment group and pioglitazone group was significantly higher than that of model group (P<0.05, P<0.05, P<0.05). There is no significant different between the Dan-gua prescription group and combination group (P=0.336).(3) GSH activity was measured in liver homogenates:There is no significant difference among the normal group, model group and treatment group (P=0.767, P=1), the content of GSH in both combination group and Dan-gua prescription group were significantly higher than that in the model group (P=0.011, P=0.002). There is no significant difference between the pioglitazone group and model group (P=0.999). Compared with normal control group, combined group and Dan-gua prescription group were significantly increased (P=0.034P=0.014).(4) The expression of SRF content in the heart:The positive expression of SRF was mainly located in nucleus.①There is negative expression of SRF in the normal group;②strong positive Brown nucleus and dense patches of staining granules can be found in model group;③Weak positive expression of SRF and scattered staining granules in brown can be found in both combination group and Dan-gua prescription group. compared with the model group, There is significant difference of SRF positive rate between combination group and Dan-gua prescription group (P<0.01);④There is positive expression of SRF in Pioglitazone group.The Pioglitazone group showed more visible brown staining granules in the nucleus but slightly weaker than the model group.3.morphology:①Control group:coronary artery intima is flat and smooth, continuous and complete. Endothelial cells is flat, most have integral structure, regularly spread, the elastic plate of tunica intima and tunica media are conherent and complete.②Model group: Coronary artery intimal was thick, unevenly breaked, the endotheial cells swellened apparently, the nucleus of the endothelium cells become larger, upright and disorganized.we can see infiltion of subcutaneous cells.③The morphology of Dan-gua prescription group and the combination group was significantly improved:most of the tunica intima of the coronary artery cannot be found thickening.④Compared with the control group, Pioglitazone group’s effect is weaker, but better than that of the untreated group:we can see thrombus and Congestion, tunica intima of the coronary artery were roughly injured,apparently thickened;of which there is hyperplasia in the smooth muscle;the elastic plates were disarranged and blured.Conclusion:1. Dan-gua prescription could decrease blood glucose, regulate lipid metabolism2.Dangua prescription has obvious antioxidant effect.3.Dangua prescription has obvious antioxidant activity. |
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