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目的:探讨雌二醇及PGE2对人子宫内膜基质细胞增殖及PCNA表达的影响。方法:采用酶消化及筛网法分离培养人子宫内膜基质细胞,免疫组化法进行细胞鉴定,将培养的细胞分为对照组(正常培养液)、雌二醇组、雌二醇+PGE2组和PGE2组,各组细胞分别作用24、48、72 h。MTT法间接测定各组细胞增殖情况,实时荧光定量PCR法测定增殖细胞内PCNA mRNA表达的变化。结果:雌二醇对细胞的增殖随着作用时间的延长而升高,各浓度之间差异无统计学意义(P>0.05)。雌二醇作用72 h,平均细胞增殖率与细胞内PCNA mRNA的相对表达量均显著高于雌二醇作用24 h,差异均有统计学意义(P<0.05)。PGE2在早期促进子宫内膜基质细胞的生长,但随着作用时间的延长细胞增殖呈现下降作用。24 h雌二醇+PGE2组与雌二醇组比较,细胞平均增殖率及PCNA mRNA的相对表达量均升高,差异均有统计学意义(P<0.05)。72h雌二醇+PGE2组与雌二醇组比较,细胞平均增殖率及PCNA mRNA的相对表达量均明显降低,差异均有统计学意义(P<0.05)。结论:雌二醇对子宫内膜细胞的增殖作用非浓度依赖性而是时间依赖性。PGE2可能参与宫腔炎症的发生,加速宫腔粘连的再形成。
Objective: To investigate the effects of estradiol and PGE2 on the proliferation of human endometrial stromal cells and the expression of PCNA. Methods: Human endometrial stromal cells were isolated and cultured by enzyme digestion and sieve method. The cells were identified by immunohistochemistry. The cultured cells were divided into control group (normal culture medium), estradiol group, estradiol + PGE2 Group and PGE2 group, the cells in each group were treated for 24, 48 and 72 h respectively. The proliferation of cells in each group was measured indirectly by MTT assay. The expression of PCNA mRNA in proliferating cells was detected by real-time fluorescence quantitative PCR. Results: The estradiol increased the proliferation of cells with the prolongation of action time. There was no significant difference between each concentration (P> 0.05). Estradiol treatment 72 h, the average cell proliferation rate and the relative expression of intracellular PCNA mRNA were significantly higher than the estradiol 24 h, the difference was statistically significant (P <0.05). PGE2 promoted the growth of endometrial stromal cells in early stage, but decreased with the extension of time. Compared with the estradiol group, the average proliferation rate and the relative expression of PCNA mRNA in 24 h estradiol + PGE2 group were significantly increased (P <0.05). 72h estradiol + PGE2 group compared with estradiol group, the average cell proliferation rate and the relative expression of PCNA mRNA were significantly decreased, the difference was statistically significant (P <0.05). Conclusion: The proliferation of estradiol on endometrial cells is not concentration-dependent but time-dependent. PGE2 may participate in the occurrence of uterine inflammation, accelerate the formation of intrauterine adhesions.