SOD在五味子乙素降低PM2.5对HTR细胞毒性中的作用

SOD is Associated with Schisandrin B Attenuating the PM2.5-Induced HTR Cells Damages

目的:探讨超氧化物歧化酶(superoxide dismutase,SOD)在五味子乙素(Schisandrin B,Sch B)降低PM2.5对人早孕胎盘绒毛膜外滋养层(human trophoblast HTR-8/SVneo,HTR)细胞毒性中的作用。方法:体外培养HTR细胞,利用小干扰RNA(small interfering RNA,siRNA)转染技术降低SOD蛋白的表达,并以不同浓度PM2.5(0~150μg/mL)处理细胞。MTS细胞增殖实验观察HTR细胞增殖。实时聚合酶链反应(real-time PCR)检测SOD mRNA的表达水平,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测SOD蛋白的表达水平。结果:SOD siRNA转染后,150μg/mL浓度的PM2.5对HTR细胞增殖的抑制率上升至33.97%,0.25、0.5、1、2μmol/L Sch B组对HTR细胞的保护率由转染前的2.74%、7.91%、10.31%、14.07%分别降至1.39%、4.98%、7.75%、9.55%。SOD siRNA转染后,0.25、0.5、1、2μmol/L Sch B组的SOD mRNA的表达量从转染前是对照组的1.26、1.47、1.86、2.25倍分别降至1.13、1.25、1.43、1.77倍。SOD siRNA转染后,0.25、0.5、1、2μmol/L Sch B组的SOD蛋白表达量从转染前是对照组的1.19、1.36、1.60、1.94倍分别降至1.09、1.16、1.27、1.41倍。结论:SOD基因的激活在Sch B降低PM2.5对HTR细胞毒性作用中起到重要作用。

Objective:To investigate whether superoxide dismutase(SOD)played a role in Schisandrin B(Sch B)attenuating the cell damage induced by PM2.5 in the human trophoblast HTR-8/SVneo(HTR)cells.Methods:With the in vitro cultured model of HTR cells,the expression of SOD gene was down-regulated by siRNA technology,and cells were treatment with PM2.5 at the doses from 0 to 150μg/mL.The cell proliferation was tested by MTS,the expression of SOD mRNA was tested by real-time PCR,and the expression of SOD protein was measured by ELISA.Results:After the expression of SOD was down-regulated by siRNA,the inhibition rate in the HTR cells induced by 150μg/mL PM2.5 was incresed to 33.97%,while the protection rates in those HTR cells induced by 0.25,0.5,1 and 2μmol/L Sch B were decresed from 2.74%,7.91%,10.31%,14.07%to 1.39%,4.98%,7.75%,9.55%respectively.Meanwhile,the expressions of SOD mRNA in the HTR cells treated with 0.25,0.5,1,2μmol/L Sch B were decreased 1.26,1.47,1.86,2.25 times than that in the cells before SOD siRNA,and decresed 1.13,1.25,1.43,1.77 times than that in the cells after SOD siRNA.Analogously,the expressions of SOD protein in the HTR cells treated with 0.25,0.5,1,2μmol/L Sch B were decreased 1.19,1.36,1.60,1.94 times than that in the cells before SOD siRNA,and decresed 1.09,1.16,1.27,1.41 times than the cells after SOD siRNA.Conclusions:The increased expression of SOD was a protective factor in the effect of Sch B attenuating the PM2.5-induced HTR cell damages.