复方精油对PM_(2.5)诱导小鼠肺微血管内皮细胞内皮间质转化的影响

Effect of compound essential oils on endothelial-mesenchymal transition of mouse pulmonary vascular endothelial cells induced by PM_(2.5)

探讨复方精油对PM_(2.5)诱导小鼠肺微血管内皮细胞(MHC)内皮间质转化(EndMT)的干预作用,并从TGF-β1/SMAD3/p-SMAD3通路分析其可能的调节机制。建立PM_(2.5)诱导小鼠MHC和复方精油干预模型,测定细胞活力和迁移能力的变化;采用RT-qPCR和Western blot检测CDH5、CD31、Col1、Acta2、S100A4和TGF-β信号通路相关因子的水平变化。结果表明复方精油可减弱PM_(2.5)对细胞迁移能力的影响;EndMT相关因子的表达随PM_(2.5)作用时间的延长呈现正向发展趋势,复方精油对PM_(2.5)诱导产生的EndMT有良好的干预作用;TGF-β1、TGF-βR1和p-SMAD3的表达均呈现下降趋势,EndMT相关因子的表达水平也有所逆转。复方精油可缓解PM_(2.5)诱导的EndMT进程,其作用与TGF-β1/SMAD3/p-SMAD3信号通路相关。

To investigate the effect of compound essential oils(CEOs)on EndMT of mouse pulmonary vascular endothelial cells(MHC)induced by PM_(2.5),and discuss the regulatory mechanism of CEOs by the TGF-β1/SMAD3/p-SMAD3 pathway.MHC cells were cultured in vitro.Design the PM_(2.5) group(500μg/mL),the CEOs group(PM_(2.5)500μg/mL+CEOs 10-5%),the inhibitor of TGF-βpathway SB431542 group(10μmol/L)and the PM_(2.5)+SB431542 group(500μg/mL+10μmol/L).The changes of cell viability and migration ability were analyzed by cell proliferation experiment and cell scratch experiment.RT-qPCR and Western blot were used to detect the levels of CDH5,CD31,Col1,Acta2,S100A4 and TGF-βsignal pathway related factors.The cell viability of MHC began to increase after 36 h in the PM_(2.5) group(P<0.0001),and there is no significant difference in the CEOs group.The cell migration increased significantly in the PM_(2.5) group,and CEOs can attenuate the effect of PM_(2.5) on cell migration(36 h,P<0.05).The result of RT-qPCR and Western blot showed that the expression of EndMT related factors showed a positive trend with the prolongation of PM_(2.5).CEOs had a beneficial intervention effect on EndMT induced by PM_(2.5) exposure.Compared with the PM_(2.5) group,Western blot showed that the expression of TGF-β1,TGF-βR1 and p-SMAD3 decreased in the CEOs group,and the expression of EndMT related factors was reversed after 48 h.CEOs ameliorates EndMT induced by PM_(2.5),which is related to TGF-β1/SMAD3/p-SMAD3 pathway.