桃红四物汤含药血清对VSMC迁移及β3整合素、MMP9和FAK%2fP38MAPK信号表达影响.pdf

桃红四物汤含药血清对 VSMC 迁移及 β3 整合素、MMP9 和 FAK/P38MAPK 信号表达的影响 研究生：易善清 导 师：武 衡 摘 要 目的：研究桃红四物汤含药血清成分对VSMC 迁移及β3 整合素、MMP 和 FAK/P38MAPK 信号表达的影响，并探讨其相关机理。 方法：制备桃红四物汤含药血清及正常大鼠血清，分离大鼠颈总动脉VSMC ， 进行体外培养、传代，观察其形态及生长特性并行免疫细胞化学鉴定，将VSMC 分为正常对照组（正常组，采用正常大鼠血清培养）与干预组（含药组，桃红四 物汤含药血清干预）分别进行处理，采用改良的Boyden chamber 方法观察两组 VSMC 迁移的活性，采用免疫细胞化学方法检测各组VSMC β3 整合素、MMP9 、 FAK 的表达的情况，采用Western blot 法检测各组不同时间及不同浓度含药血清 干预下VSMC P38MAPK 表达的变化。 结果：体外成功培养出大鼠颈总动脉VSMC ，经形态观察及免疫细胞化学鉴 定证实为平滑肌细胞，正常对照组VSMC 存在迁移情况，桃红四物汤含药血清 干预后大鼠VSMC 迁移细胞数减少、迁移距离缩短（P0.01 ），VSMC 在桃红 四物汤含药血清干预后相对于正常对照组，β3 整合素、MMP9 、FAK 及P38MAPK 的表达均降低（P0.01 ），P38MAPK 的表达随含药血清干预时间的延长及作用 浓度的增加而降低（P0.01 ）。 结论：桃红四物汤含药血清可抑制VSMC 的迁移，且桃红四物汤含药血清 对 β3 整合素、MMP 和FAK/P38MAPK 信号表达均产生负性影响，此作用参与 了对VSMC 的迁移及增殖的抑制作用。 关键词 桃红四物汤含药血清，VSMC ，细胞迁移，β3整合素，MMP ，FAK ， P38MAPK 7 Effect of blood serum containing THSWT on vascular smooth muscle cells’ migration and the expression of integrin β3,MMP9 and FAK/38MAPK signal postgraduate: Yishanqing advisor: Wuheng Abstract Objective: To research the effects of blood serum containing Decoction of TaoHong SiWuTang(THSWT) on vascular smooth muscle cells(VSMC) migration and the expression of integrin β3, matrix metalloproteinase9(MMP9) and focal adhesion kinase(FAK)/P38 MAPK signal ，and to investigate the mechanisms about it. Methods: Blood serum containing Decoction of THSWT and normal blood serum from rats were Prepared. The vascular smooth muscle cells were isolated from common carotid artery of rats, then were cultured in vitro, transfered of culture and identified by immunocytochemistry. Vascular smooth muscle cells were disposed to be cultured with normal