| Objectives To observe the effect of Shizidaiping formula(SZDP) on cell apoptosis, insulin secretion, the expression of BCL-2 and Caspase3 in MIN6 cell under the high glucose and free fatty acid condition, to explore the protection effect of SZDP in MIN6 cell and the related mechanism.Methods 1 Selected 45 SPF male GK rats,T2 DM model was established by feeding high fat feed for 4 weeks after adaptive feeding 1 week, detected 2h plasma glucose(PG2h), PG2h>11.1 mmol·L-1 was considered succeed T2 DM model. And the T2 DM rats were randomly divided into model group, metformin group, SZDP groups with high, medium and low dose(SZDP-H, SZDP-M and SZDP-L) respectively. The normal group and model group were administered intragastrically with purified water. Other groups were administered intragastrically with corresponding drugs for 8 weeks. The levels of PG2 h and FBG were detected at the beginning of treatment, and at the end of the 8th week. Oral glucose tolerance test(OGTT)was detected on 0 min,60 min,120min,calculated the area under the curve(AUC).Hb Alc was detected at the end of 8th week. The rats were sacrificed after 8 week, detected the level of TG, CHO, HDL-c, LDL-c, Fins, C-P, SOD, MDA, and calculated islet cell function index(HOMA-?).Took GK rat pancreas, histopathological examination was stained by HE,and then detect Bcl-2 and Caspase3 expression with Western blot method.2 To establish MIN6 cell damage model by glucose(21mmol?L-1)+ palmitic acid(0.37mmestablishol?L-1) and then divided the MIN6 cell into four groups, they are the control group, model group, SZDP group with different concentration and metformin group. The cell activity was examined by cck-8; the insulin secretion were measured by ELISA;the rate of apoptosis were measured by Annexin VFITC&PI and the level of Bcl-2 and Caspase3 were examined by Western blotting after SZDP Intervention.Results 1 compared with model group, SZDP can reduce the levels of FBG and PG2 h, each does group has significant different(P<0.05,P<0.01). Compared with the model group, the content of Hb Alc in SZDP group was significantly decreased(P<0.05). Compared with model group, AUC result in 4 weeks, SZDP-H, SZDP-M and SZDP-L group have no significant difference(P>0.05), no differences were found in 8 weeks compared with model group(P>0.05), but has a trend to decrease AUC. SZDP-H,SZDP-M and SZDP-L group can reduce the levels of TG, and significant differences compared with model group(P<0.01), SZDP-H group compared with model group can reduce the level of CHO and LDL-c(P<0.05), and HDL-c, each dose group has no statistical difference(P > 0.05) compared with model group, but has a trend to improve HDL-c. SZDP group C-P, FINS, and HOMA-? level has no statistical difference compared with model group(P > 0.05), but has a trend to increase the level of FINS and C-P and improve HOMA-?. SZDPH, SZDP-M and SZDP-L group can significantly increase the levels of SOD and reduce the levels of MDA compared with model group(P<0.05,P<0.01), The HE staining results showed that the islet morphology of SZDP group was more regularly than model group, and the number of islet cells increased. The level of BCL-2 was improved and the level of Caspase3 was decreased after SZDP treatment(P<0.01).2 SZDP can significantly improve MIN6 cells' activity under high glucose and free fatty acid damage(P<0.05), decrease the rate of apoptosis, increase the level of insulin stimulated by high glucose in cell supernatant(P<0.01), decrease the level of insulin stimulated by low glucose in cell supernatant(P<0.01), improve the expression level of BCL-2 and decrease the expression level of Caspase3(P<0.05,P<0.01).Conclusions 1 SZDP can improve glucose and lipid metabolism in T2 DM rats to a certain extent; 2 SZDP can protect pancreatic islet cells by inhibiting the early apoptosis of islet cells and regulating insulin secretion; 3 One of the mechanisms of SZDP inhibition of cell apoptosis may be realized by increase BCL-2 expression and reduce Caspase3 expression; 4 The protective effect of SZDP on islet cells may be related to the inhibition of oxidative stress. |
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