菩人丹改善高糖波动状态下INS-1细胞胰岛素分泌功能的分子机制  

Purendan Improving the Molecular Mechanism of Ins-1 Cell Insulin Secretion under High Glucose Fluctuation

作　　者：白颖慧 杜子亮[2] 陈书[1] 鲁碧楠[1] 庞宗然[1] 

机构地区：[1]中央民族大学中国少数民族传统医学研究院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 [2]北京市回民医院,北京100053

出　　处：《中国中医基础医学杂志》2016年第12期1620-1624,1637,共6页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金资助项目(81072963)-菩人丹超微粉调控PTP1B蛋白与IRS2/PI3K/Akt信号转导改善胰岛β细胞形态与功能的作用机制;国家自然科学基金青年基金项目(81302946)-基于IR信号转导系统的菩人丹修管T2DM胰岛β细胞损伤作用靶点研究

摘　　要：目的:考察菩人丹改善高糖波动状态下INS-1细胞胰岛素分泌功能的分子机制。方法:将INS-1细胞置于含33.3 mmol·L-1和11.1 mmol·L-1葡萄糖RPMI1640培养液内12 h交替培养,持续培养5 d构建高糖波动细胞模型。分别以5%、10%菩人丹含药血清对高糖波动状态下INS-1细胞干预24 h,并以二甲双胍含药血清作为阳性对照。正常对照组细胞以含11.1 mmol·L-1葡萄糖的RPMI1640培养液进行培养。采用CCK-8试剂盒检测细胞活力,大鼠胰岛素ELISA试剂盒测定胰岛素分泌量,Western blot分析IRS2、Akt、m TOR、P70s6k和PTP1B的蛋白质表达水平及磷酸化水平。结果:与正常组比较,高糖波动能够显著降低INS-1细胞活力及胰岛素分泌量,下调INS-1细胞的IRS2蛋白质表达水平,提高IRS2磷酸化水平,降低INS-1细胞Akt、m TOR和P70s6k的磷酸化水平,上调PTP1B蛋白质表达水平。与模型组比较,菩人丹含药血清能增加INS-1细胞活力及胰岛素分泌量,上调IRS2蛋白质表达,抑制IRS2磷酸化,促进Akt、m TOR和P70s6k的磷酸化,下调PTP1B蛋白表达水平。结论:菩人丹调控高糖波动诱导的INS-1细胞IRS2、Akt、m TOR、P70s6k和PTP1B的蛋白质表达水平及磷酸化水平,改善高糖波动状态下胰岛β细胞胰岛素分泌功能。Objective： To study the molecular mechanism of Pu Ren Dan improving the secretion function of INS-1 cell under fluctuated high glucose. Methods： INS-1 cell was cultured in high glucose RPMI1640 medium containing 33. 3 mmol·L-1glucose and conventional RPMI1640 medium containing 11. 1 mmol·L-1glucose each 12 hours for 5 days to induce fluctuated high glucose cell model. INS-1 cell was intervened in 5% and 10% drug serum containing PRD for 24 hours as PRD experimental group,drug serum containing Metformin as the positive control group. The INS-1 cell in the normal control group was cultured in RPMI1640 medium containing 11. 1 mmol·L-1glucose. Using CCK-8 kit to detect cell viability,ELISA to test the insulin secretion of INS-1 cell, Western blot assay to test proteins expression and phosphorylation levels of IRS2,Akt,m TOR,P70s6 k,PTP1B. Results： Compared with the control group,cell viability and insulin secretion level of INS-1 cell in FG decreased significantly. Protein expression level of IRS2 decreased,phosphorylation of IRS-2 increased; phosphorylations of Akt,m TOR and P70s6 k lowered significantly; while the protein expression level of PTP1 B increased. Compared with the model group,cell viability and insulin secretion level of INS-1 cell in PRD experimental group increased significantly. Protein expression level of IRS2 increased,phosphorylation of IRS2lowered; phosphorylations of Akt,m TOR and P70s6 k increased significantly; while the protein expression level of PTP1 B decreased. Conclusion： Pu Ren Dan regulates the protein expression and phosphorylation levels of IRS2,Akt,m TOR,P70s6 k and PTP1 B of INS-1 cell induced by fluctuated high glucose,improving the insulin secretion function of INS-1 cell under fluctuated high glucose.

关 键 词：INS-1细胞 高糖波动 胰岛素分泌 菩人丹 IRS2-PI3K/Akt 

分 类 号：R285.5[医药卫生—中药学]