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目的观察不同雌、孕激素水平对子宫内膜上皮细胞Claudin-4表达的影响。方法 RL95-2细胞系培养于含10%胎牛血清培养基中,在培养基中加入雌、孕激素,一组孕酮的浓度恒定为0.025μmol/L,雌二醇终浓度分别为1、0.5、0.25、0.125、0.062 5和0 nmol/L;另一组雌二醇的浓度恒定为0.25 nmol/L,孕酮终浓度分别为0.1、0.05、0.025、0.012 5、0.006 25和0μmol/L。两组分别培养24、48和72 h,RT-PCR和Western印迹法检测RL95-2细胞的Claudin-4基因和蛋白表达。结果在雌激素恒定的条件下,RL95-2细胞的Claudin-4基因和蛋白表达随着孕激素水平的升高而升高(P<0.01),随着培养时间的延长,其表达Claudin-4基因和蛋白水平升高(P<0.01)。在孕激素恒定的条件下,RL95-2细胞的Claudin-4基因和蛋白表达随着雌激素水平的升高而降低(P<0.01),随着培养时间的延长,其表达Claudin-4基因和蛋白水平降低(P<0.01)。结论 Claudin-4在子宫内膜细胞的表达水平与雌孕激素的水平具有密切的联系,Claudin-4可能参与子宫容受性的形成和胚泡在子宫的着床有关。
Objective To observe the effect of different estrogen and progesterone levels on the expression of Claudin-4 in endometrial epithelial cells. Methods RL95-2 cells were cultured in medium containing 10% fetal bovine serum. Estrogen and progesterone were added to the medium. The concentration of progesterone in a group of cells was 0.025μmol / L, the final concentration of estradiol was 1, 0.5, 0.25, 0.125, 0.062 5 and 0 nmol / L respectively; the other group of estradiol was constant at 0.25 nmol / L and the final progesterone concentrations were 0.1, 0.05, 0.025, 0.012 5, 0.006 25 and 0 μmol / . The two groups were cultured for 24, 48 and 72 h respectively. The expression of Claudin-4 gene and protein in RL95-2 cells were detected by RT-PCR and Western blot. Results Claudin-4 gene and protein expression of RL95-2 cells were increased with the increase of progesterone level (P <0.01) under constant estrogen condition. With the prolongation of culture time, the expression of Claudin-4 Gene and protein levels increased (P <0.01). Under the condition of constant progesterone, Claudin-4 gene and protein expression of RL95-2 cells decreased with the increase of estrogen level (P <0.01). With the prolongation of culture time, the expression of Claudin-4 gene and Protein levels decreased (P <0.01). Conclusions The expression of Claudin-4 in endometrial cells is closely related to the levels of estrogen and progesterone. Claudin-4 may be involved in the formation of uterine tolerance and the implantation of blastocysts in the uterus.