中文摘要：

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。

英文摘要：

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.