激活TRPV1对自发性高血压大鼠血管平滑肌细胞增殖的影响刘赟.DOC

激活TRPV1对自发性高血压大鼠血管平滑肌细胞增殖的影响 刘 赟，李敬诚，尹延伟，黎炳护，郭 露，廖少琼，曹小洁，张明杰，周毅，陈磊，龙春燕，张莉莉 （400042 重庆，第三军医大学大坪医院神经内科） [摘 要] 目的 探讨辣椒素激活瞬时受体电位香草醛亚家族1 (transient receptor potential vanilloid 1, TRPV1) 对体外培养的自发性高血压大鼠(spontaneously hypertensive rats，SHR) 血管平滑肌细胞 (vascular smooth muscle cells，VSMCs) 增殖的影响。方法 用组织贴块法分别培养SHR大鼠及其正常对照Wistar Kyoto (WKY)大鼠的主动脉血管平滑肌细胞，细胞免疫荧光鉴定VSMCs。将两种来源的细胞分为对照组、辣椒素 (capsaicin, TRPV1激动剂) 浓度梯度组 (0.1μmol/L、1μmol/L、10μmol/L)、辣椒素(1μmol/L) +5’-iodo-resiniferatoxin (TRPV1拮抗剂, iRTX, 1μmol/L) 组。CCK-8检测VSMCs增殖情况；Western blot检测TRPV1蛋白、磷酸化-Akt蛋白（p-Akt）和总Akt（t-Akt）蛋白表达情况。结果 ①WKY、SHR来源的细胞中均有TRPV1表达，SHR-VSMC中TRPV1蛋白表达低于WKY-VSMC；辣椒素干预后，WKY-VSMC、SHR-VSMC中TRPV1表达均显著升高（P＜0.05）。②SHR-VSMC增殖率高于WKY-VSMC（P0.05）；TRPV1激动剂辣椒素呈剂量依赖性地抑制SHR-VSMC的增殖（P＜0.05）；TRPV1拮抗剂iRTX则使得辣椒素抑制细胞增殖的作用被阻断（P＜0.05）。③SHR-VSMC中p-Akt蛋白表达高于WKY-VSMC（P＜0.05）；辣椒素激活TRPV1使VSMCs中p-Akt表达下降（P＜0.05），TRPV1拮抗剂iRTX可抑制此效应（P＜0.05）。结论 激活TRPV1可能通过下调p-Akt蛋白表达抑制SHR-VSMC增殖。 [关键词] TRPV1；自发性高血压大鼠；血管平滑肌细胞；细胞增殖 [中图法分类号] R322.121；R329.28；R544.1 [文献标志码] A [基金项目] 重庆市杰出青年科学基金 (CSTC2012JJJQ10003)。 [通讯作者] 张莉莉，电话：(023E-mail： HYPERLINK mailto:zll197312@ zll197312@ Effect of TRPV1 on proliferation of vascular smooth muscle cells in spontaneously hypertensive rats Liu Yun, Li Jingcheng, Li Binghu, Yin Yanwei, Guo Lu, Liao Shaoqiong, Cao Xiaojie, Zhang Mingjie,Zhou Yi,Chen Lei, Long, Chunyan, Zhang Lili ( Department of Neurology ,Institute of Suregery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China) [Abstact] Objective To observe the effect of transient receptor potential vanilloid 1(TRPV1) on the proliferation of vascular smooth muscle cells (VSMCs) in spontaneously hypertensive rats (SHR). Methods VSMCs from SHR and their age-matched Wistar Kyoto rats (WKY) were cultured from aortas using the tissue expant method. VSMCs identified by morphology and immunofluorescence. Then, VSMCs were incubated in the absence or presence of TRPV1 agonist capsaicin at gradient concentratio