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加味三棱丸含药血清对子宫内膜异位症患者子宫内膜细胞雌激素分泌的抑制作用及其对细胞凋亡的影响

[Inhibitory effect of Jiawei Sanleng pill medicated serum on estrogen production and its influence on apoptosis of human endometrial cells of endometriosis].

作者信息

Li Ao, Wang Ying, Xu Xiao-Yu

机构信息

College of Chemistry and Bioengineering, Chongqing University of Technology, Chongqing 400050, China.

出版信息

Zhong Yao Cai. 2010 Mar;33(3):401-6.

Abstract

OBJECTIVE

To study the inhibitory effect on estradiol (E2) production of endometriosis by medicated serum of Jiawei Sanleng pill (SLW) and approach the mechanism of inducing apoptosis of human endometrial cells of endometriosis by medicated serum of SLW.

METHODS

The eutopic endometrial cells of hysteromyoma and endometriosis patients were cultured. Taking that of the endometrial cells of hysteromyoma as control, the secretion level of E2 of endometrial cells in the culture media supernatant at different time point with the treatment of high, middle and low dose of SLW serum was detected by electrochemiluminescence immunoassay and the apoptosis percentage of endometrium cells in early stage and late stage was detected by flow cytometry and TUNEL method. After the optimal time for SLW to induce apoptosis of eutopic endometrial cells of endometriosis in early stage and late stage was identified based on time-effect relationship, another endometrium cells were divided into six groups: hysteromyoma endometrium group, eutopic endometrium of endometriosis group, eutopic endometrium of endometriosis + middle dose of SLW serum group, eutopic endometrium of endometriosis + anastrozole serum group, eutopic endometria of endometriosis + middle dose of SLW serum + E2 group and eutopic endometrium of endometriosis + E2 group. The apoptosis percentage of endometrial cells in early stage and late stage of each group was detected according to the optimal time point respectively.

RESULTS

The secretion level of E2 of eutopic endometrium of endometriosis was decreased by SLW, which showed the dependence of concentration and time. The apoptosis percentage of endometrial cells in hysteromyoma endometrium group in early stage and late stage was increased along with the time and they was significantly higher than that of eutopic endometrium of endometriosis group at the same time point (P < 0.05 or P < 0.01). After 72 hours and 96 hours, with the treatment of middle dose of SLW serum, the apoptosis percentage of eutopic endometrial cells of endometriosis in early stage and late stage was 35.67 +/- 1.73 and 57.56 +/- 1.09 respectively, and the effect of inducing apoptosis was significantly stronger than that of the other time point. After 72 hours, with the combined treatment of middle dose of SLW serum and 5.30 ng/L E2, or after 96 hours, with the combined treatment of middle dose of SLW serum and 6. 36 ng/L E2, the apoptosis percentage of eutopic endometrial cells in early stage and late stage was decreased to 11.25 +/- 2.00 and 7.72 +/- 0.71 respectively, which had significant difference with that of middle dose of SLW serum group (P < 0.01). The apoptosis of the eutopic endometrial cells of endometriosis was induced by medicated serum of SLW could not be completely counteracted by E2 add-back treatment. Just with the treatment of 5.30 ng/L or 6.36 ng/L E2, the apoptosis percentage of eutopic endometrial cells of endometriosis in early stage and late stage tended to decrease, but there was no statistical significance (P > 0.05). Anastrozole, a specific aromatase inhibitor, had the contrary action to E2.

CONCLUSION

SLW could decrease the secretion of E2 so as to induce apoptosis of the eutopic endometrial cells of endometriosis.

摘要

目的

研究加味三棱丸(SLW)含药血清对子宫内膜异位症雌二醇(E2)分泌的抑制作用,并探讨SLW含药血清诱导子宫内膜异位症患者子宫内膜细胞凋亡的机制。

方法

培养子宫肌瘤和子宫内膜异位症患者的在位子宫内膜细胞。以子宫肌瘤患者的子宫内膜细胞为对照,采用电化学发光免疫分析法检测高、中、低剂量SLW血清作用于不同时间点时,培养基上清中子宫内膜细胞E2的分泌水平,采用流式细胞术和TUNEL法检测早期和晚期子宫内膜细胞的凋亡率。根据时间-效应关系确定SLW诱导子宫内膜异位症在位子宫内膜细胞早期和晚期凋亡的最佳时间后,将另一批子宫内膜细胞分为6组:子宫肌瘤子宫内膜组、子宫内膜异位症在位子宫内膜组、子宫内膜异位症在位子宫内膜+中剂量SLW血清组、子宫内膜异位症在位子宫内膜+阿那曲唑血清组、子宫内膜异位症在位子宫内膜+中剂量SLW血清+E2组、子宫内膜异位症在位子宫内膜+E2组。分别根据最佳时间点检测各组早期和晚期子宫内膜细胞的凋亡率。

结果

SLW降低了子宫内膜异位症在位子宫内膜E2的分泌水平,呈浓度和时间依赖性。子宫肌瘤子宫内膜组早期和晚期子宫内膜细胞凋亡率随时间增加,且在同一时间点显著高于子宫内膜异位症在位子宫内膜组(P<0.05或P<0.01)。72小时和96小时后,中剂量SLW血清作用下,子宫内膜异位症在位子宫内膜细胞早期和晚期凋亡率分别为35.67±1.73和57.56±1.09,诱导凋亡作用明显强于其他时间点。72小时后,中剂量SLW血清与5.30 ng/L E2联合处理,或96小时后,中剂量SLW血清与6.36 ng/L E2联合处理,早期和晚期在位子宫内膜细胞凋亡率分别降至11.25±2.00和7.72±0.71,与中剂量SLW血清组相比有显著差异(P<0.01)。SLW含药血清诱导的子宫内膜异位症在位子宫内膜细胞凋亡不能被E2回补处理完全抵消。仅给予5.30 ng/L或6.36 ng/L E2处理时,子宫内膜异位症在位子宫内膜细胞早期和晚期凋亡率有下降趋势,但无统计学意义(P>0.05)。特异性芳香化酶抑制剂阿那曲唑对E2有相反作用。

结论

SLW可降低E2分泌,从而诱导子宫内膜异位症在位子宫内膜细胞凋亡。

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