菩人丹调控IRS2、BAD、FOXO1蛋白表达和磷酸化抑制INS-1细胞凋亡的分子机制  

Effects of Pu-Ren-Dan Formula on the Apoptosis of INS-1 Cell through Regulating Expressions of IRS2,BAD and FOXO1 and Its Mechanism

作　　者：陈书[1] 鲁碧楠[1] 杜子亮[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：《中国中医基础医学杂志》2013年第11期1341-1344,1356,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金资助项目(81072963)

摘　　要：目的:阐明菩人丹抑制软脂酸诱导INS-1细胞凋亡的作用,并从分子水平揭示其作用机制。方法:构建软脂酸(PA,0.5mmol/L)损伤INS-1细胞模型,给予菩人丹含药血清干预24 h后,采用CCK-8法、Hoechst荧光染色法、流式Annexin V-FITC/PI双染法、caspase-3和caspase-8活性检测试剂盒检测INS-1细胞凋亡;利用Western blot法分析IRS2、BAD和FOXO1的蛋白质表达及磷酸化水平。结果:PA可显著降低INS-1细胞活力,升高caspase-3、-8活性,诱导细胞凋亡。菩人丹则抑制INS-1细胞凋亡,增加细胞活力,上调IRS2表达,下调FOXO1表达,促进BAD和FOXO1磷酸化,降低IRS2磷酸化。结论:菩人丹通过调控胰岛素信号传导系统中凋亡相关蛋白IRS2、BAD、FOXO1的表达和磷酸化,抑制PA诱导的胰岛β细胞凋亡,发挥保护胰岛β细胞的作用。Objective： To clarify the inhibition effect of PRD on the apoptosis of INS-1 cell in palmitic acid condition, and explore the molecular mechanism. Method： Experimental injured INS-1 cell models were induced by palmitic acid （containing 0.5 mmol/L） , and intervened 24 h by PRD drug-serum. Cell viability, INS-1 cell apoptosis and expression levels of related proteins and its phosphorylations were detected by CCK-8 assay, Hoechst 33342 staining, Flow Cytometry and AnnexinV-FITC/PI staining, caspase-3, -8 Activity Assay Kit and Western Blotting assay, repeetively. Results： Palmitic acid can decrease cell viability, increase activities of easpase-3 and caspase-8, and induce INS-1 cell apoptosis. After PRD treatment, protein expressions of IRS2, BAD, and FOXO1 were increased, while protein expressions of FOXOI and phosphorylations of IRS2 were reduced significantly. Conclusion： PRD drug-serum shows a protective effect on apoptosis of islet 13 cell which induced by palmitic acid, and attains the objective by regulating the expression level of related proteins and its phosphorylations in insulin signaling system.

关 键 词：菩人丹 INS-1细胞 软脂酸 细胞凋亡 分子机制 

分 类 号：R285.5[医药卫生—中药学]