DNA甲基转移酶1在一氧化氮合酶抑制剂诱导的滋养细胞自噬中的作用

Role of DNMT1 in trophoblast autophagy induced by nitric oxide synthase inhibitors

目的探讨DNA甲基转移酶1(DNMT1)在一氧化氮合酶抑制剂L-NAME诱导的胎盘滋养细胞自噬中的作用。方法 L-NAME干预滋养细胞后,Western blot检测LC3BⅡ/Ⅰ和p62的表达;qRT-PCR和Western blot检测DNMT1的表达水平;另外,干扰和过表达DNMT1后,检测LC3BⅡ/Ⅰ和p62的表达水平。结果与对照组比较,L-NAME组LC3BⅡ/Ⅰ蛋白表达增加,而p62表达显著降低(P <0.05);且DNMT1表达降低(P <0.05)。另外,干扰DNMT1后,LC3BⅡ/Ⅰ蛋白表达升高,而p62降低(P <0.05);相反,过表达DNMT1后,LC3BⅡ/Ⅰ蛋白表达降低,而p62表达增加。结论 DNMT1能够抑制L-NAME诱导的胎盘滋养细胞发生自噬。

Objective To investigate the effect of DNA methyltransferase 1(DNMT1)on autophagy of placental trophoblast cells induced by nitric oxide synthase inhibitor(L-NAME).Methods Trophoblast cells were treated with L-NAME.The expression of LC3BⅡ/Ⅰand p62 protein was detected by Western blot and that of DNMT1 by qRT-PCR and Western blot.In addition,after DNMT1 silenced and over-expressed respectively,the expression of LC3BⅡ/Ⅰand p62 protein were detected.Results Compared with that in the control group,the ratio of LC3BⅡ/Ⅰwas increased,while p62 and DNMT1expression reduced in L-NAME group(P﹤0.05).In addition,the ratio of LC3BⅡ/Ⅰwas increased and p62 expression significantly reduced after DNMT1 knock-down.On the contrary,the expression of LC3BⅡ/Ⅰprotein was decreased,while the p62 protein was increased.Conclusion DNMTI can inhibit the autophagy of placental trophoblast cells induced by L-NAME.