五味子乙素对BaP致HTR8-SVneo细胞损伤的保护作用及机制

The Protection Effect and Mechanism of Schisandrin B on Extravillous Trophoblast Cell Line Damage Caused by BaP

目的:研究五味子乙素(Schisandrin B,Sch B)预防苯并芘(Ba P)致人绒毛膜外滋养层细胞(HTR8-SVneo)损伤的作用机制。方法:通过新型四唑化合物(MTS)法观察和测定,最终选取0.5,1,2μmol/L Sch B分别作用HTR8-SVneo细胞24 h后再给予20μmol/L Bap染毒处理24 h,MTS检测各组细胞生长情况;JC-1荧光染色法观察各组线粒体膜电位荧光强度变化;酶联免疫吸附试验(ELISA)检测各组细胞内促凋亡蛋白(Bax)、细胞色素C(cytochromec,cyt-c)蛋白浓度;蛋白质印迹法(Western blotting)检测各组的凋亡蛋白(Caspase-9)。结果:1Ba P染毒组OD值为1.187±0.015,0.5,1,2μmol/L Sch B预保护24 h后的OD值分别为1.483±0.022、1.489±0.048和1.531±0.240,差异有统计学意义(P<0.05)。2Ba P染毒组线粒体荧光强度弱,0.5,1,2μmol/L Sch B预保护24 h后的荧光强度较强,差异有统计学意义(P<0.05)。3Ba P染毒组Bax、cyt-c蛋白浓度升高,0.5,1,2μmol/L Sch B预保护24 h后细胞内Bax、cyt-c蛋白浓度降低,差异有统计学意义(P<0.05)。4Ba P染毒组细胞内caspase-9蛋白表达升高,0.5,1,2μmol/L Sch B预保护24 h后细胞内caspase-9蛋白表达降低,差异有统计学意义(P<0.05)。结论:Sch B可以预防Ba P致HTR8-SVneo细胞损伤,其作用机制可能与抗线粒体途径有关。

Objective：To explore the protection mechanism of Schisandrin B on extravillous trophoblast cell line damage caused by Ba P. Methods：After 0.5,1,2 μmol/L Sch B deal with human HTR8-SVneo cells 24 h,added 20 μmol/L Bap 24 h,using MTS measured cells growth. Used JC-1 fluorescent staining to measure the change of mitochondrial membrane potential in each group. Respectively used ELISA to measure the concentration of Bax and cyt-c protein in each group. Respectively used Western blotting to analyse the expression of caspase-9 in each group. Results： 1MTS results showed that Ba P alone cell OD was 1.187±0.015,0.5,1,2 μmol/L Sch B the role of OD was 1.483 ±0.022,1.489±0.048 and 1.531 ±0.240,compared with Ba P alone group,the differences were statistically significant（P〈0.05）. 2JC-1 fluorescent staining results showed that Ba P alone cell mitochondrial fluorescent intensity was lower and 0.5,1,2 μmol/L Sch B the role of mitochondrial fluorescent intensity was higer. Compared with Ba P alone group,the differences were statistically significant（P〈 0.05）. 3 ELISA results showed that Ba P alone cell the concentration of Bax,cyt-c protein were higher and 0.5,1,2 μmol/L Sch B the role of the concentration of Bax,cyt-c protein were lower,respectively. Compared with Ba P alone group,the differences were statistically significant（P〈0.05）. 4 Western blotting results showed that Ba P alone cell the concentration of caspase-9 protein was higer,and 0.5,1,2μmol/L Sch B the role of the concentration of caspase-9 protein was lower,respectively. Compared with Ba P alone group,the differences were statistically significant（P〈0.05）. Conclusions：Schisandrin B can prevent damage of extravillous trophoblast cell line caused by Ba P,whose mechanism may be associated with resistance to mitochondrial pathway.