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  

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

作　　者：杜子亮[1] 鲁碧楠[1] 陈书[1] 白永飞[1] 德力格玛[1] 庞宗然[1] 

机构地区：[1]中国少数民族传统医学国家民委-教育部重点实验室,北京100081

出　　处：《中国实验方剂学杂志》2013年第22期173-177,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072963)

摘　　要：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） 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.

关 键 词：菩人丹 高糖 大鼠胰岛素瘤细胞系细胞 凋亡 Bal-xl Bcl-2相关死亡促进因子 叉头框转录因子O亚族1 

分 类 号：R285.5[医药卫生—中药学]