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作 者:刘志能[1] 姚珍薇[1] 唐良萏[1] 卞度宏[1]
机构地区:[1]重庆医科大学附属第一医院妇产科,重庆400016
出 处:《生殖与避孕》2008年第2期84-88,124,共6页Reproduction and Contraception
摘 要:目的:探讨全反式维甲酸(ATRA)对人异位子宫内膜间质细胞间隙连接细胞间通讯(GJIC)、connexin基因、蛋白的调节作用及意义。方法:取24例子宫内膜异位症患者卵巢处异位内膜标本,分离纯化得到间质细胞,分别用0.1mmol/L、1mmol/L、10mmol/LATRA处理24h、48h、72h、96h、120h,同时设置空白对照组,采用荧光漂白后恢复技术检测异位内膜间质细胞的GJIC功能,并检测与GJIC功能密切相关的Cx43、Cx26、Cx32的基因及蛋白表达。结果:ATRA处理72h内,1mmol/L、10mmol/L组的异位内膜间质细胞荧光恢复功能明显增强;0.1mmol/LATRA组的异位内膜间质细胞荧光恢复功能无明显变化。未经ATRA处理的异位内膜间质细胞中无Cx43、Cx26、Cx32的mRNA及蛋白表达;经1mmol/LATRA处理后,异位内膜间质细胞中检测到Cx43mRNA和蛋白表达,但未见Cx26、Cx32的mRNA及蛋白表达。结论:ATRA能选择性地诱导Cx43基因及蛋白表达,从而有效上调异位内膜间质细胞的GJIC功能;ATRA可能是治疗子宫内膜异位症的潜在药物。Objective: To explore the effects of all-trans-retinoic acid (ATRA) on functional gap junctional intercellular communication (GJIC), connexin genes and proteins in human endometriotic stromal cells. Methods: Stromal cells were isolated and purified from 24 endometriotic tissues located in ovaries of patients undergoing surgery for endometriosis. Endometriotic stromal cells had been treated with 0.1 μmol/L, 1 μmol/L and 10 μmol/L ATRA for 24 h, 48 h, 72 h, 96 h and 120 h, respectively, control groups were introduced simultaneously. Fluorescence recovery alter photobleaching was applied to determine the function of GJIC in strorml cells. Cx43, Cx26 and Cx32, which are associated with GJIC, were investigated at the mRNA and protein levels. Results: The ability of fluorescence recovery was enhanced significantly in endometriotic stromal cells exposed to 1 μmol/L and 10 μmol/L ATRA for different time periods, up to 72 h. No evidence for significant fluorescence recovery was confirmed in endometriotic stromal cells treated with 0.1 μmol/L ATRA. Cx43, Cx26 and Cx32 were not detected at mRNA levels and protein levels in control group. Treatment of endometriotic stromal cells with 1 μmol/L ATRA for 72 h induced the expression of Cx43 mRNA and protein while lost the same effects on Cx26 and Cx32. Conclusion: ATRA induced selectively the expressions of Cx43 mRNA and protein in ectopic endometrial stromal cells, then GJIC in these cells was effectively up-regulated, which implied ATRA may be a potential drug to administer the patients suffering from endometriosis.
关 键 词:全反式维甲酸(ATRA) 间质细胞 间隙连接细胞间通讯(GJIC) 子宫内膜异位症(EMS) 荧光漂白后恢复技术
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