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目的探讨过氧化物酶体增殖物激活受体(peroxisome proliferator activated receptor gamma,PPAR-γ)激动剂罗格列酮(rosiglitazone,RSG)对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导小鼠血管平滑肌细胞(vascular smooth muscle cells,VSMCs)表型转化的抑制作用及其可能的机制。方法体外培养小鼠血管平滑肌细胞,应用AngⅡ诱导VSMCs的表型转化,给予罗格列酮预处理后用Transwell检测细胞迁移并在激光共聚焦显微镜下观察细胞微丝骨架,用RT-q PCR和Western blot分别检测PPAR-γ、PKGⅠα以及VSMCs收缩型标志因子(smooth muscle 22 alpha,SM22α)m RNA和蛋白水平。应用PPAR-γ抑制剂预处理后再给予罗格列酮刺激,Western blot检测PKGⅠα和SM22α蛋白水平。结果 1罗格列酮(20μmol/L)预处理可抑制AngⅡ诱导VSMCs的迁移(P<0.05)。2罗格列酮(20μmol/L)预处理后可抑制AngⅡ诱导VSMCs微丝骨架的形成。3RT-q PCR检测结果显示:罗格列酮(20μmol/L)预处理后可抑制AngⅡ诱导PKGⅠα和SM22αm RNA的下调作用(P<0.05)。4Western blot检测结果显示:罗格列酮(20μmol/L)预处理后可抑制AngⅡ诱导PKGⅠα和SM22α蛋白水平的下调作用(P<0.05);PPAR-γ抑制剂GW9662可减弱罗格列酮对PKGⅠα和SM22α的促表达作用(P<0.05)。结论罗格列酮通过激活PPAR-γ来抑制AngⅡ诱导VSMCs的表型转化,激活PPAR-γ可能通上调PKGⅠα表达发挥上述作用。
Objective To investigate the effect of peroxisome proliferator activated receptor gamma (PPAR-γ) agonist rosiglitazone on vascular smooth muscle cells To investigate the inhibitory effect of vascular smooth muscle cells (VSMCs) phenotype transformation and its possible mechanism. Methods Vascular smooth muscle cells (VSMCs) were cultured in vitro. The phenotype of VSMCs was induced by AngⅡ. After pretreatment with rosiglitazone, the migration of VSMCs was detected by Transwell and the actin cytoskeleton was observed by laser scanning confocal microscopy. Western blot was used to detect the mRNA and protein levels of PPAR-γ, PKGⅠα and VSMCs smooth muscle 22 alpha (SM22α), respectively. PPAR-γ inhibitor pretreatment before being given rosiglitazone stimulation, Western blot detection PKG Ⅰ α and SM22α protein levels. Results 1 Rosiglitazone (20 μmol / L) pretreatment inhibited the migration of VSMCs induced by AngⅡ (P <0.05). 2 Rosiglitazone (20μmol / L) pretreatment can inhibit Ang Ⅱ induced VSMCs microfilament scaffold formation. 3RT-q PCR results showed that rosiglitazone (20μmol / L) pretreatment inhibited the down-regulation of PKGⅠα and SM22αmRNA induced by AngⅡ (P <0.05). Western blot showed that rosiglitazone pretreatment inhibited the down-regulation of PKGⅠα and SM22α protein levels induced by AngⅡ (P <0.05); PPAR-γ inhibitor GW9662 attenuated the effect of rosiglitazone on PKGⅠα And SM22α (P <0.05). Conclusion Rosiglitazone inhibits the phenotypic transformation of VSMCs induced by AngⅡ by activating PPAR-γ. Activation of PPAR-γ may up-regulate PKGⅠα expression.