左旋炔诺孕酮对人子宫内膜细胞体外增殖及细胞间通讯的影响

Effect of Levonorgestrel on Proliferation and Gap Junction Intercellular Communication of Human Endometrial Cell

【目的】探讨左旋炔诺孕酮对体外培养的人子宫内膜腺上皮细胞及间质细胞增殖、间隙连接细胞间通讯(gap junction intercellular communication,GJIC)及连接蛋白(connexin,Cx)43、32表达的影响。【方法】采用MTT法观察LNG对人子宫内膜细胞的增殖调节作用,采用划痕染料标记示踪技术,观察左旋炔诺酮(levonorgestrel,LNG)对原代培养的人子宫内膜腺上皮细胞及间质细胞GJIC的影响,采用激光共聚焦显微镜扫描技术,观察LNG对Cx43、Cx32表达的影响。【结果】LNG浓度为5×10-5mol/L时,对子宫内膜间质和上皮细胞均有抑制生长作用(P<0.05),体外培养的人子宫内膜间质细胞的GJIC功能较腺上皮细胞强(P<0.05),LNG可显著提高人子宫内膜腺上皮细胞及间质细胞间荧光黄染料的扩散(P<0.05);同种Cx腺上皮细胞表达较间质细胞强(P<0.05),LNG对两种细胞Cx的表达没有影响(P>0.05)。【结论】LNG可抑制人子宫内膜腺上皮细胞及间质细胞体外增殖,显著促进细胞GJIC功能,提示其在逆转子宫内膜增殖性病变的阶段扮演重要角色;LNG未影响子宫内膜细胞Cx43、Cx32的表达,提示Cx表达量不能完全反映GJIC功能。

[Objective] To explore the effects of levonorgestxel on proliferation, gap junction intercellular communication and Cx43, Cx32 expression of stromal and glandular cells in vitro. [Methods] The effects of levonorgestrel （LNG） on the proliferation of endometrial stromal and glandular cells were detected by MTT （3-（4, 5- dimethylthiazol-2-yl）-2, 5-diphenyltetrazolium bromide） method. The scrap loading dye transfer was used to investigate the effects of LNG on GJIC of endometrial stromal and glandular cells. The influence of levonorgestrel （LNG） on Cx43 and Cx32 expression was investigated with a laser scanning confocal microscope. [Results] 5× 10^-5 mol/L LNG significantly depressed the proliferation of endometrial stromal and glandular cells. The ability of GJIC function was higher in the endometrial stromal cells than that of the glandular cells in vitro. 5×10^-5 mol/L LNG leading to greater GJIC after 48h （P〈0.05）. Cx32 expressions in endometrial stromal and glandular cells were significantly higher than Cx43. 5×10^-5 mol/L LNG had no effect on Cx expression. [Conclusions] LNG could inhibit the proliferation of endometrial stromal and glandular cells in vitro. The function of LNG＇s upregulation on GJIC of endometrial stromal and glandular cells may be one of mechanisms which invert endometrial hyperplasia. LNG had no effect on Cx expression of endometrial stromal and glandular cells, which indicates Cx expression could not completely reflect GJIC function in vitro.