摘要
目的 :探讨 HIV- 1蛋白酶抑制剂 nelfinavir对胰岛 β细胞内胰岛素信号传导蛋白磷酸化的影响。方法 :体外观察鼠胰岛素瘤 INS- 1细胞经 nelfinavir治疗 4 8h后 ,用 10 0 nmol/ L胰岛素刺激 2 min,细胞裂解产物中的胰岛素信号传导蛋白磷酸化采用免疫沉淀和 Western印迹方法测定。用台盼蓝染色细胞计数和 MTT衰减试验 ,评估nelfinavir对 INS- 1细胞活力的影响。结果 :nelfinavir降低胰岛素刺激的胰岛素受体底物 IRS- 2和 Akt- Thr3 0 8磷酸化 ,并呈剂量依赖关系。nelfinavir浓度为 10 μmol/ L时 ,分别降低 IRS- 2和 Akt- Thr3 0 8磷酸化达 5 2 %和 5 5 % ,在这个浓度下 nelfinavir没有细胞毒性作用。结论 :nelfinavir治疗可以损害胰岛 β细胞内 IRS-
Objective: To investigate whether HIV-1 protease inhibitor nelfinavir alters the insulin-stimulated phosphorylation of insulin signaling parameters in rat insulinoma INS-1 cells. Methods: INS-1 cells were incubated with nelfinavir for 48 h and stimulated with 100 nmol/L insulin for 2 min. Immunoprecipitation and Western blot analysis of the insulin-stimulated insulin receptor substrate (IRS)-1,-2 and Akt-Thr^(308)-phosphorylation were performed on cell lysates. Cytotoxic effects of nelfinavir were measured by cell count with trypan blue and MTT reduction test. Results: Nelfinavir decreased insulin-stimulated phosphorylation of IRS-2 and Akt-Thr^(308) in a dose-dependent manner;for 10 μmol/L of nelfinavir,the decrease was 52% and 55%,respectively. Conclusion: Treatment with nelfinavir might impair IRS-2-mediated signaling in pancreatic β cells.
出处
《浙江大学学报（医学版）》
CAS
CSCD
2004年第4期311-314,共4页
Journal of Zhejiang University(Medical Sciences)
基金
德国国家研究基金 (KI5 0 3/ 7- 3)资助项目
