大鼠cx43基因干扰载体构建及对星形胶质细胞谷氨酸释放的影响.docVIP

组蛋白H3、H4乙酰化及H3K4甲基化对人妊娠子宫平滑肌细胞PRA/PRB的影响 罗 慧，陈 诚，梁志清（400038 重庆，第三军医大学西南医院妇产科） [摘要]目的通过组蛋白去乙酰化酶抑制剂（trichostatin A,TSA）、H3K4甲基化酶抑制剂(5’-deoxy-5’-methylthioadenosine, MTA)处理人妊娠子宫平滑肌细胞，干预组蛋白H3、H4乙酰化及H3K4甲基化水平，探讨组蛋白H3、H4乙酰化及H3K4甲基化对人妊娠子宫平滑肌细胞PRA/PRB的影响。 方法 分离纯化人妊娠子宫平滑肌细胞(n=16)，Western bolt检测孕激素受体(progesterone receptor, PR)的两种亚型——孕激素受体A(progesterone receptor A, PRA)、孕激素受体B(progesterone receptor B, PRB)的表达情况。分别利用不同浓度TSA、MTA对其进行处理，RT-qPCR检测PR、PRA、PRB的表达；染色质免疫共沉淀技术(Chromatin immunoprecipitation, ChIP)技术比较处理前后PRA、PRB启动子区H3、H4乙酰化及H3K4三甲基化水平。 结果 TSA可使PRA/PRB明显增高(P0.05)，使PRA启动子区H3、H4乙酰化水平明显上升(P0.05)。MTA可使PRA/PRB明显下降(P0.05)，PRA启动子区H3K4乙酰化水平明显下降(P0.05)。两种药物的干预主要是通过对PRA的调控来调节PRA/PRB比值。 结论 组蛋白H3、H4乙酰化和H3K4甲基化均可使人妊娠子宫平滑肌细胞PRA/PRB比值发生改变，可能参与“功能性孕激素撤退”机制的调节。 [关键词] 组蛋白乙酰化；H3K4甲基化；妊娠；子宫平滑肌细胞 [中图法分类号]R394-33；R394.3；R34 [文献标志码]A Effects of histone acetylation and H3K4 methylation on PRA/PRB in human uterine smooth muscle cells during pregnancy Luo Hui, Chen Cheng, Liang Zhiqing (Department of Gynecology and Obstetrics, Southwest Hospital, Third Military Medical University, Chongqing, 400038, China) [Abstract] Objective To determine the effects of histone acetylation and H3 lysine 4 (H3K4) methylation on the ratio of A and B isoforms of the human progesterone receptor (PRA/PRB) in human uterine smooth muscle cells during the course of pregnancy. Methods Human uterine smooth muscle cells of pregnancy were isolated from 16 full-term pregnant women receiving caesarean section, and then purified by collagenase digestion. The expressions of PRA and PRB were detected by western blotting in the cells and in the cells treated with different concentrations of histone deacetylase inhibitor, trichostatin A (TSA) or H3K4 methyltransferase inhibitor, 5’-Methylthioadenosine (MTA). The mRNA levels of progesterone receptor (PR), PRA and PRB were detected by RT-qPCR. Chromatin immunoprecipitation (ChIP) was used to analyze acetylation of histones H3 and H4, and trimethylation of H3K4 in the