组蛋白精氨酸甲基化酶在人体外受精发育阻滞胚胎中的功能研究  

Functional Study of Protein Arginine Methyltransferase in In Vitro-Fertilized Human Embryos with Developmental Arrest

作　　者：黄雪梅[1] 樊军 马蓉宁[1] 江酉琼[1] 冷媚[1] 叶飞[1] HUANG Xue-mei;FAN Jun;MA Rong-ning;JIANG You-qiong;LENG Mei;YE Fei(Reproductive Medicine Center,Sichuan Provincial Maternity and Child Health Care Hospital,Chengdu 610045,China)

机构地区：[1]四川省妇幼保健院生殖医学中心,成都610045

出　　处：《南昌大学学报（医学版）》2022年第6期6-10,共5页Journal of Nanchang University：Medical Sciences

基　　金：四川省自然科学基金(20190115)。

摘　　要：目的探究组蛋白精氨酸甲基化酶(PRMT)在体外受精的人类胚胎发育阻滞中的作用。方法收集丢弃的70枚体外受精人类胚胎,选取12枚正常发育的胚胎为正常组,12枚发育阻滞胚胎为阻滞组。蛋白印迹和免疫荧光检测2组组蛋白精氨酸甲基化酶6(PRMT6)蛋白质水平,蛋白印迹检测组蛋白H3在精氨酸2的二甲基化(H3R2me2)和组蛋白H3在赖氨酸4的三甲基化(H3K4me3)水平。敲除46枚发育阻滞胚胎中的PRMT6(PRMT6敲除组,n=46),qPCR和蛋白印迹检测阻滞组和PRMT6敲除组的PRMT6 RNA、PRMT6蛋白、H3R2me2和H3K4me3水平,多能性标记Oct4、Nanog和Sox2,观察阻滞组和PRMT6q敲除组胚胎培养7 d的情况。结果与正常组相比,阻滞组的PRMT6蛋白质水平显著增加[(1.160±0.054)比(1.530±0.13)],H3R2me2升高[(1.340±0.019)比(1.710±0.011)],H3K4me3水平下降[(1.650±0.030)比(1.220±0.015)],差异有统计学意义(P<0.05)。与阻滞组比较,PRMT6敲除组PRMT6 RNA[(1.660±0.029)比(1.230±0.017)]、PRMT6蛋白[(1.400±0.030)比(1.050±0.058)]及H3R2me2蛋白水平显著下调[(1.410±0.011)比(1.340±0.019)],而H3K4me3水平升高[(1.710±0.011)比(1.340±0.019)];多能性标记Oct4[(1.710±0.016)比(1.160±0.049)]、Nanog[(1.690±0.089)比(1.260±0.051)]和Sox2[(1.680±0.091)比(1.310±0.057)]的水平被下调,差异有统计学意义(P<0.05)。结论敲除PRMT6可以挽救部分发育阻滞的胚胎,甚至单个发育受阻的胚胎也可以发育成融合胚胎。敲除PRMT6可促进人体外受精发育阻滞胚胎的发育。Objective To investigate the role of protein arginine methyltransferase(PRMT)in in vitro-fertilized human embryos with developmental arrest.Methods Seventy discarded in vitro-fertilized human embryos were collected,including 12 normally developed embryos(normal group)and 12 developmentally arrested embryos(arrested group).The protein levels of PRMT6 were detected by Western blotting and immunofluorescence.The dimethylation of histone H3 at arginine 2(H3R2me2)and the trimethylation of histone H3 at lysine 4(H3K4me3)were measured by Western blotting.In addition,PRMT6 was knocked out in developmentally arrested embryos(PRMT6 knockout group,n=46).The levels of PRMT6 RNA,PRMT6 protein,H3R2me2,H3K4me3,Oct4,Nanog and Sox2 were determined by qPCR and Western blotting.Furthermore,the embryos were observed after culture for 7 days.Results Compared with the normal group,the developmentally arrested embryos showed an increase in PRMT6 protein(1.160±0.054 vs 1.530±0.130)and H3R2me2(1.340±0.019 vs 1.710±0.011),along with a decrease in H3K4me3(1.650±0.030 vs 1.220±0.015)(P<0.05).Compared with the arrested group,PRMT6 knockout decreased the levels of PRMT6 RNA(1.660±0.029 vs 1.230±0.017),PRMT6 protein(1.400±0.030 vs 1.050±0.058),H3R2me2(1.410±0.011 vs 1.340±0.019),Oct4(1.710±0.016 vs 1.160±0.049),Nanog(1.690±0.089 vs 1.260±0.051)and Sox2(1.680±0.091 vs 1.310±0.057),but increased the level of H3K4me3(1.710±0.011 vs 1.340±0.019)(P<0.05).Conclusion PRMT6 knockout can rescue developmentally arrested embryos,and even single developmentally arrested embryos can develop into fused embryos.Therefore,PRMT6 knockout can promote the development of in vitro-fertilized human embryos with developmental arrest.

关 键 词：人体外受精 组蛋白精氨酸甲基化酶 组蛋白精氨酸甲基化酶6敲除 发育阻滞胚胎 

分 类 号：R714.8[医药卫生—妇产科学]