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目的:观察菩人丹(Puren Dan,PRD)对高糖诱导的INS-1细胞凋亡、Bal-xl/Bcl-2相关死亡促进因子(BAD)和叉头框转录因子O亚族1(FOXO1)蛋白表达的影响,探讨PRD恢复INS-1细胞分泌功能的相关分子机制。方法:采用血清药理学方法制备菩人丹含药血清,建立高浓度葡萄糖(33.3 mmol·L-1)诱导INS-1细胞损伤模型,以菩人丹药物血清干预24 h;实验分为5组,即正常对照组(control组)、高糖模型组(HG组)、菩人丹药物血清高剂量组(H-PRD组)、菩人丹药物血清低剂量组(L-PRD组)和二甲双胍药物血清对照组(MF组);采用CCK8试剂盒检测细胞活力,流式Annexin V-FITC/PI双染法考察细胞凋亡水平,Western blotting assay检测BAD和FOXO1蛋白表达水平及其磷酸化水平。结果:与正常组比较高糖能够显著降低INS-1细胞活力、诱导细胞凋亡、降低INS-1细胞BAD,FOXO1丝氨酸磷酸化水平;与模型组比较菩人丹含药血清(终体积分数10%)则能抑制INS-1细胞凋亡,增加细胞活力,下调FOXO1表达,促进BAD和FOXO1丝氨酸磷酸化。结论:菩人丹药物血清能够减少高糖诱导的INS-1细胞凋亡,这种作用与上调BAD和FOXO1蛋白丝氨酸磷酸化水平直接相关。
Objective: To observe the effect of Puren Dan (PRD) on the apoptosis of INS-1 cells induced by high glucose, the expression of Bal-xl / Bcl-2 related death promoting factor (BAD) and forkhead box transcription factor O subfamily 1 ) Protein expression, explore the PRD restore INS-1 cell secretory function related molecular mechanism. Methods: Bodam Dan serum was prepared by serum pharmacology. The injury model of INS-1 cells induced by high concentration of glucose (33.3 mmol·L-1) was established. The serum was injected for 24 h. The experiment was divided into 5 groups (Control group), high glucose group (HG group), Bodhidong drug serum high dose group (H-PRD group), Bodam Dan drug serum low dose group (L-PRD group) and metformin (MF group). The viability of cells was detected by CCK8 kit and the apoptosis rate was analyzed by flow cytometry Annexin V-FITC / PI double staining. The expression of BAD and FOXO1 protein and the phosphorylation of Bcl-2 protein were detected by Western blotting assay. Results: Compared with the normal group, high glucose could significantly reduce the INS-1 cell viability, induce apoptosis and decrease the serine phosphorylation of BAD and FOXO1 in INS-1 cells. Compared with the model group, %) Inhibited INS-1 cell apoptosis, increased cell viability, down-regulated FOXO1 expression, and promoted serine phosphorylation of BAD and FOXO1. Conclusion: BUDAMAN serum can reduce high glucose-induced apoptosis of INS-1 cells, which is directly related to the up-regulation of serine phosphorylation of BAD and FOXO1.