摘要
目的 :探讨胰岛素样生长因子 1(IGF 1)促血管平滑肌细胞 (VSMC)增殖的细胞内信号转导机制。方法 :体外培养的兔血管平滑肌细胞分 3组处理 ,以细胞计数、噻唑盐比色法测定细胞增殖能力 ,以磷脂酰肌醇 3激酶 (PI3 K)特异性抑制剂渥漫青霉素 (WT)孵育细胞间接反映PI3 K作用。WesternBlot定量磷酸酶PTEN表达水平 ,免疫沉淀、特异底物diC16PIP3 绿色试剂法测定PTEN脂质磷酸酶活性。结果 :IGF 1(10 0 μg/L)使细胞计数及MTT比色A值分别增加至对照组的 2 .8倍和 3.8倍 ,WT抑制VSMC增殖 ,并完全逆转IGF 1的作用 (均P <0 .0 1)。各浓度IGF 1对PTEN蛋白表达水平无明显影响 ,其对PTEN活性的抑制呈浓度 (10～ 10 0 μg/L)及时间 (3min～ 2 4h)依赖性 (均P <0 .0 1)。结论 :IGF 1促VSMC增殖作用与活化PI3
Aim: To investigate the cellular signal transduction pathway of vascular smooth muscle cell (VSMC) proliferation stimulated by insulin-like growth factor-1(IGF-1).Methods: Rabbit aortic VSMCs was cultured in 3 groups. Cell proliferating ability was determined by measuring cell number and mitochondrial dehydrogenase(MD) activity(MTT assay).Wortmannin(WT),the specific inhibitor of phosphatidylinositol 3-kinase(PI_3K), was used to evaluate indirectly the possible involvment of PI_3K. Western blotting was used to detect the protein expression of phosphatase PTEN. Dephosphate action of PTEN on its specific substrate diC16PIP_3 was measured by green reagent method.Results: IGF-1(100 μg/L) increased cell number and MD activity by 2.8～3.8 fold. WT markedly inhibited VSMC proliferation and completely abolished the above effects of IGF-1. IGF-1 inhibited PTEN activity in a concentration-(10～100 μg/L) and time-(3 min～24 h) dependent manner(P<0.01). Conclusion: IGF-1 increases VSM proliferation by increasing PI_3K activity and inhibiting PTEN activity.
出处
《中国应用生理学杂志》
CAS
CSCD
北大核心
2004年第3期259-262,共4页
Chinese Journal of Applied Physiology
关键词
磷脂酰肌醇3-激酶
PTEN
胰岛素样生长因子-1
血管平滑肌细胞
phosphatidylinositol 3-kinase
phosphotase and tensin homology deleted on chromosome ten (PTEN)
insulin-like growth factor-1
vascular smooth muscle cell
