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目的:研究加味三棱丸(SLW)对子宫内膜异位症(内异症)雌二醇(E2)生成的抑制作用,并探讨其诱导内异症内膜细胞凋亡的机制。方法:培养子宫肌瘤和内异症患者在位子宫内膜细胞,以子宫肌瘤内膜细胞为对照,内异症在位内膜细胞经含SLW大鼠血清高、中、低剂量处理后,分别在不同的时间点采用电化学发光免疫法检测E2的分泌水平;流式细胞术和TUNEL法检测细胞的早期和晚期凋亡率;通过时效关系考察,明确SLW诱导内异症在位内膜细胞早、晚期凋亡的最佳时间后,重新将培养的内膜细胞分为6组:子宫肌瘤组、内异症组、内异症+SLW血清中剂量组、内异症+阿那曲唑血清组、内异症+SLW血清中剂量+雌二醇组和内异症+雌二醇组,在最佳时间点检测各组细胞的早期和晚期凋亡率。结果:SLW可显著降低内异症在位内膜细胞分泌E2的水平,并呈浓度和时间依赖性。子宫肌瘤组内膜细胞的早、晚期凋亡百分率随时间延长而增加,并显著高于同时间点的内异症组(P<0.05或P<0.01);经SLW血清中剂量处理72、96 h后,内异症在位内膜细胞的早、晚期凋亡百分率分别为(35.67±1.73)%和(57.56±1.09)%,较其他处理时间的诱导作用显著;当SLW血清中剂量与雌二醇5.30 ng/L联合处理72 h或与雌二醇6.36 ng/L联合处理96 h后,细胞的早、晚期凋亡百分率分别降低至(11.25±2.00)%和(7.72±0.71)%,与SLW血清中剂量比较,差异显著(P<0.01),添加的雌二醇浓度未能完全抑制SLW诱导内异症内膜细胞的凋亡;单独给予雌二醇5.30、6.36 ng/L进行处理,内异症内膜细胞的早、晚期凋亡百分率有进一步降低的趋势,但无统计学意义(P>0.05),芳香化酶抑制剂阿那曲唑与雌二醇的作用恰好相反。结论:SLW可能通过降低内异症在位内膜细胞分泌雌二醇的水平,发挥其诱导内膜细胞凋亡的作用。
Objective: To study the inhibitory effect of Modified Sanling Pills (SLW) on estradiol (E2) production in endometriosis and to explore its mechanism of endometriosis-induced endometrial apoptosis. Methods: The uterine leiomyoma and endometriosis patients were cultured in endometrial cells. The uterine leiomyoma endometrial cells were used as control. Endometriosis cells of endometriosis were treated with high, medium and low dose of SLW serum , And the secretion of E2 was detected by electrochemiluminescence immunoassay at different time points. Flow cytometry and TUNEL method were used to detect the early and late apoptosis rate of cells; Membrane cells early and late apoptosis of the best time, re-culture of endometrial cells were divided into 6 groups: uterine fibroid group, endometriosis group, endometriosis + SLW serum dose group, endometriosis + A Naztezole serum group, endometriosis + SLW serum medium dose + estradiol group and endometriosis + estradiol group. At the best time point, the early and late apoptosis rate of each group of cells was detected. RESULTS: SLW significantly reduced the level of E2 secreted by endometriotic cells in endometriosis, in a concentration- and time-dependent manner. The percentage of early and late apoptosis of endometrial cells in uterine myoma group increased with time and was significantly higher than that in endometriosis group at the same time point (P <0.05 or P <0.01) After 96 h, the percentage of early and late apoptotic cells in eutopic endometriosis cells were (35.67 ± 1.73)% and (57.56 ± 1.09)%, respectively, which were significantly higher than those in other treatment groups. The percentage of early and late apoptotic cells were decreased to (11.25 ± 2.00)% and (7.72 ± 0.71)% respectively after treated with estradiol 5.30 ng / L for 72 h or estradiol 6.36 ng / L for 96 h, (P <0.01). The concentration of estradiol added did not completely inhibit the apoptosis of endometriotic cells induced by SLW. Administration of estradiol alone at 5.30,6.36 ng / L (P> 0.05), the effect of the aromatase inhibitor anastrozole and estradiol was just the opposite. Conclusion: SLW may play an important role in inducing apoptosis of endometrial cells by decreasing the level of endometrial cells secreting estradiol in endometriosis.