菩人丹对高糖诱导的INS-1细胞凋亡、BAD和FOXO1蛋白表达的影响

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。

Effect of Puren Dan Recipe Containing Drug-serum on the Apoptosis of INS-1 Cell and Expressions of BAD and FOXO1 in High Glucose Condition

Objective: To observe the effect of Puren Dan (PRD) recipe on the apoptosis of INS-1 cell and expression of Bal-xl/Bcl-2 associated death promoter (BAD) and forkhead box O1 (FOXO1) in high glucose condition, and explore the molecular mechanism of PRD on restoring secretion function of INS-1 cell. Method: Experimental injured INS-1 cell model was induced by high glucose (containing 33.3 mmol·L^-1), and intervened by PRD drug-serum. INS-1 cells were divided into 5 groups:the control group, high glucose group, high PRD drug-serum group, low PRD drug-serum group and metformin drug-serum group. Cell viability, INS-1 cell apoptosis and expression level of related proteins and its phosphorylation were detected by CCK-8 assay, flow cytometry and AnnexinV-FITC/PI staning and Western blotting assay, respectively. Result: High glucose could decrease cell viability and induce INS-1 cell apoptosis, and it also downregulatedf phosphorylation level of BAD and FOXO1. PRD can increase cell viability, reduce apoptosis of INS-1 cell downregulated expression of FOXO1 and improved phosphorylation level of BAD and FOXO1. Conclusion: PRD drug-serum shows a protective effect on apoptosis INS-1 cell which induced by high glucose, and the mechanism may be associated with increase of the phosphorylation levels of BAD and FOXO1.