apelin13促大鼠血管平滑肌细胞增殖的pi3kakt信号通路研究 word格式.docxVIP

Apelin-13 促大鼠血管平滑肌细胞增殖的PI3K/Akt 信号通路研究研究生：李芳指 导 教 师：陈临溪 教授摘要目的: 研究 G 蛋白偶联受体 APJ 的内源性配体多肽 apelin-13 是否通过 PI3K/Akt 信号通 路促进血管平滑肌细胞(vascular smooth muscle cells，VSMCs)的生长增殖，发现 apelin-APJ 系统促血管平滑肌细胞增殖的新的信号转导机制。方法: 培养大鼠胸主动脉血管平滑肌细胞，噻唑蓝比色法（MTT）观察 PI3K 抑制剂 LY294002 和 Akt 抑制剂 1701-1 对 apelin-13 促 VSMCs 增殖的影响；Western blot 检测 PI3K、 pPI3K、pAkt、pERK1/2、ERK1/2、细胞周期蛋白 D1(cyclin D1)等的表达。结果: MTT 法显示 PI3K 抑制剂 LY294002 和 Akt 抑制剂 1701-1 能明显抑制 apelin-13 诱 导的 VSMCs 增殖;Western blot 检测结果显示，apelin-13（0 、0.5、1、2、4 μmol/L）刺 激大鼠血管平滑肌细胞 pPI3K、pAkt 表达增加，以 1μM 最为明显；1μM apelin-13 分别 刺激大鼠 VSMCs 0、 5、 15、 30、 45、 60min，pPI3K、pAkt 表达增加，在 30min 时最为明显; PI3K 阻断剂 LY294002 明显抑制 apelin-13 诱导的 PI3K、Akt、ERK1/2 的磷 酸化及 cyclinD1 的表达，Akt 阻断剂 1701-1 明显抑制 apelin-13 诱导的 Akt、ERK1/2 的 磷酸化及 cyclinD1 的表达。结论: Apelin-13 通过 PI3K/Akt 信号转导通路促进大鼠血管平滑肌细胞增殖。关键词: Apelin；APJ；血管平滑肌细胞；PI3K；Akt；ERK1/2；cyclinD1;The study of PI3K/Akt signaling transduction pathway of rat vascular smooth muscle cells proliferation induced by apelin-13AbstractAIM: To observe whether G protein-coupled receptor APJ endogenous ligand apelin-13 induce vascular smooth muscle cells (VSMCs) proliferation via PI3K/Akt signaling transduction pathway, and find new signaling transduction pathway of apelin/APJ.Methods: VSMCs were prepared from thoracic aortas of male Sprague-Dawley rats by the explant method. The effect of PI3K inhibitor LY294002 and Akt inhibitor 1701-1 on VSMCsproliferation were measured by MTT assay. Expression of PI3K、pPI3K、pAkt、ERK、pERK1/2and cyclin D1 were detected by Western blotting.Results: MTT assay showed that PI3K inhibitor LY294002 and Akt inhibitor 1701-1 significantly inhibited the VSMCs proliferation induced by apelin-13. Western blotting analysis showed that apelin-13 phosphorylated PI3K and Akt in dose and time dependent.PI3K inhibitor LY294002 significantly decreased PI3K、Akt、ERK1/2 phosphorylation andthe expression of cyclinD1 induced by apelin-13; The Akt inhibitor significantly diminished Akt、ERK1/2 phosphorylation and the