胰岛素通过活性氧的产生促进VEGF表达及血管平滑肌细胞迁移和增殖  

Insulin stimulates ROS production to promote VEGF expression,cells migration and proliferation of vascular smooth muscle cells

作　　者：梅爱红[1] 刘俊许[2] 陈思锋[2] 孟丹[2] 

机构地区：[1]同济大学附属第十人民医院诊断学教研室,上海200072 [2]复旦大学上海医学院生理与病理生理学系,上海200032

出　　处：《中国病理生理杂志》2013年第2期272-277,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30600245;o.81170298);上海市卫生局科研项目(No.c106629)

摘　　要：目的:探讨活性氧(reactive oxygen species,ROS)在胰岛素促进的血管平滑肌细胞迁移和增殖中的作用及分子机制。方法:采用原代培养的大鼠主动脉血管平滑肌细胞,应用DCF-DA荧光探针检测细胞内ROS的生成;应用实时定量PCR、Western blotting和ELISA法检测mRNA和蛋白的表达;应用转染报告基因的方法检测基因的转录活性;划痕法测定细胞迁移;CCK-8法测定细胞增殖。结果:胰岛素处理后血管平滑肌细胞内ROS产生明显增加。过氧化氢酶和NADPH氧化酶抑制剂二亚苯基碘鎓(DPI)明显抑制胰岛素促进的ROS生成及p-Akt、p-p70S6K1和p-ERK1/2蛋白的表达。过氧化氢酶和DPI明显降低胰岛素促进的血管内皮生长因子(vascular endothelial growth factor,VEGF)的mRNA和蛋白表达及转录激活。抑制ROS产生明显抑制胰岛素刺激的血管平滑肌细胞迁移和增殖。结论:胰岛素通过NADPH氧化酶途径促进血管平滑肌细胞ROS产生。ROS介导了胰岛素促进的Akt/p70S6K1和ERK信号通路的激活、VEGF表达及血管平滑肌细胞的迁移和增殖。AIM： To determine the role and mechanism of reactive oxygen species （ROS） in the migration and proliferation of insulin-stimulated vascular smooth muscle ceils （VSMCs）. METHODS： Primary VSMCs were isolated from the rat thoracic aortas and cultured in vitro. ROS production was measured by DCF-DA staining. Real-time PCR, Western blotting, and ELISA were employed to detect the mRNA expression and protein level of vascular endothelial growth factor （VEGF）. The gene transcriptional activity was determined by reporter assay. Cell migration and proliferation were measured by wound-healing and CCK-8 assays. RESULTS： Insulin stimulated intracellular ROS production in cultured rat VSMCs, which was inhibited by catalase and NADPH oxidase inhibitor diphenylene iodonium （DPI）. Catalase and DPI in- hibited insulin-induced the activation of Akt/p70S6K1 and ERK1/2. Insulin increased the expression of VEGF. Treatment with catalase and DPI inhibited insulin-induced VEGF transcriptional activation and expression of VEGF at mRNA and pro- tein levels. Catalase and DPI significantly inhibited insulin-stimulated VSMC migration and proliferation. CONCLUSION： Insulin stimulates ROS production through NADPH oxidase in VSMCs. ROS mediates insulin-stimulated Akt/p7OS6K1 and ERK1/2 pathways, VEGF expression, and VSMC migration and proliferation.

关 键 词：胰岛素 活性氧 血管平滑肌细胞 血管内皮生长因子 细胞增殖 细胞迁移 

分 类 号：R363.2[医药卫生—病理学]