全反式维甲酸上调异位子宫内膜间质细胞GJIC功能的机制

Mechanism of up-regulation on gap junctional intercellular communication by all-trans retinoic acid in human endometriotic stromal cells in vitro

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)功能的调节作用及其机制.方法:取人子宫内膜异位症(EMs)患者的间质细胞,采用0.1,1,10μmol/L ATRA于24,48,72,96和120 h分别进行处理,同时设置空白对照组.采用激光共聚焦显微镜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和Cx32的基因及蛋白表达.结果:经ATRA处理后,1μmol/L与10μmol/L处理组的异位内膜间质细胞荧光恢复功能明显增强;0.1μmol/L处理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和Cx32的mRNA及蛋白表达;经1,10μmol/LATRA处理后,异位内膜间质细胞中检测到Cx43 mRNA和蛋白表达,但未见Cx26和Cx32的mRNA及蛋白表达.间质细胞的GJIC功能与去磷酸化Cx43蛋白的表达有关.结论:ATRA能选择性的诱导Cx43基因及其去磷酸化蛋白的表达,从而上调异位内膜间质细胞的GJIC功能;ATRA酸可能是治疗EMs的潜在药物.

AIM： To explore the possible mechanism underlying the up-regulation on gap junctional intercellular communication （GJIC） by all-transretinoic acid （ATRA） in human endometriotic stromal cells. METHODS： Ectopic endometrial stromal cells isolated from human endometriotic tissues were treated with 0. 1, 1 and 10 μmol/L ATRA for 24, 48, 72, 96 and 120 h, respectively, and control groups were introduced simultaneously. Laser scanning confocal microscope was applied to determine the function of GJIC in these stromal cells. Cx43, Cx26 and Cx32, which are associated with GJIC, were investigated at the mRNA and protein levels. RE- SULTS ： The ability of fluorescence recovery was enhanced signifi- cantly in endometriotic stromal cells exposed to 1 and 10 tLmol/L ATRA. No evidence for fluorescence recovery was confirmed in en- dometriotic stromal cells treated with 0.1 μmol/L ATRA. Cx43, Cx26 and Cx32 were not detected at mRNA and protein levels in control groups. Treatment of endometriotic stromal cells with 1, 10μmol/L ATRA for 72 h induced the expressions of Cx43 mRNA and protein while lost the same effects on Cx26 and Cx32. The function of GJIC in endometriotic stromal cells was only associated with dephosphorylated Cx43 protein. CONCLUSION： ATRA induces selectively the expressions of Cx43 mRNA and dephosphorylated Cx43 protein in endometriotic stromal cells, and then GJIC in these cells are effectively up-regulated, which implys that ATRA may be a potential drug to administer the patients suffering from endometriosis.