加味三棱丸抗子宫内膜异位症内膜细胞雌二醇生成机制的研究

Study on inhibitory effect of medicated serum of SLW on estrogen production by human endometrial cells of endometriosis

目的:探讨加味三棱丸(SLW)抗子宫内膜异位症内膜雌激素生成的作用机制。方法:以SLW含药血清作用于体外培养的内异症在位内膜细胞,RT-PCR检测子宫肌瘤组、给予SLW含药血清前后内异症组类固醇类产生因子-1(steroidgenic factor-1,SF-1)、小鸡卵清蛋白上游启动转录因子(chicken ovalbumin upstream-transcription factor,COUP-TF),17-β-羟甾类脱氢酶1(17-beta-hydroxysteroid dehydrogenase 1,17-β-HSD 1)和17-β-羟甾类脱氢酶2(17-beta-hydroxysteroid dehydrogenase 2,17-β-HSD 2)mRNA的表达。Western blot检测上述各组SF-1和COUP-TF蛋白的表达。结果:子宫肌瘤对照组SF-1,17-β-HSD 1 mRNA及SF-1蛋白的表达明显低于内异症内膜组,而COUP-TF,17-β-HSD 2 mRNA及COUP-TF蛋白的表达明显高于内异症在位内膜组,差异有极显著性(P<0.01)。以内异症在位内膜为对照,给予对照组SLW 5.0,2.5 g.kg-1.d-1大鼠含药血清(体积分数为0.1)作用48 h后,COUP-TF mRNA和蛋白、17-β-HSD 2 mRNA的表达明显高于对照组,而SF-1 mRNA和蛋白、17-β-HSD 1 mRNA的表达明显低于对照组,差异有显著性或极显著性(P<0.05或P<0.01)。SLW 1.25 g.kg-1.d-1组COUP-TF mRNA和蛋白的表达水平明显高于对照组,SF-1蛋白和17-β-HSD 1 mRNA表达明显低于对照组(P<0.01),而SF-1,17-β-HSD 2 mRNA的表达却无明显改变。结论:加味三棱丸可降低子宫内膜异位症在位内膜细胞分泌雌二醇的水平,与其抑制SF-1和17-β-HSD 1,增强COUP-TF和17-β-HSD 2的表达密切相关。

Objective： To explore the mechanism of inhibitory effect of SLW on estrogen production by endometrial cells of endometriosis. Method： After the model of eutopic primary cultured endometrial cells of endometiosis and hysteromyoma in vitro was successfully established, the changes of steroidgenic factor-1（SF-1）, chicken ovalbumin upstream-transcription factor（COUP-TF）,17-beta-hydroxysteroid dehydrogenase 1（17-β-HSD1）and 17-beta-hydroxysteroid dehydrogenase 2 （17-β-HSD 2）mRNA were detected by RT-PCR before and after treatment of medicated serum of SLW. The changes of SF-1 and COUP-TF protein were also observed by western blot synchronously according to the same treatment method mentioned-above. Meanwhile ,the data of hysteromyoma group was obtained from the above experiments. Result： The expression of SF-1 mRNA and protein ,17-β-HSD1 mRNA was weak,but COUP-TF mRNA and protein, 17-β-HSD2 mRNA was remarkable in Hysteromyoma endometrium,as compared with those of endometiosis ,which was taken as control group（P〈0.01）.After the 48 hours’ treatment of medicated serum of 5. 0,2.5 g·kg^-1·d^-1 of SLW ,the expression of COUP-TF mRNA and protein,17-β-HSD 2 mRNA was found significantly increased, but SF-1 mRNA and protein,17-β-HSD 1 mRNA was decreased in contrast to the control group（P〈0. 01 or P〈0. 05）. Although the expresson of COUP-TF mRNA and protein was increased,SF-1 protein and 17-β-HSD 1 mRNA was decreased in 1.25 g·kg^-1·d^-1 medicated serum group ,compared with those of the control group（P〈0.01）,the low dose group had no apparent inhibitory effect on the expression of SF-1,17-β-HSD 2 mRNA. Conclusion： The medicated serum of SLW could inhibit the secretion of estradiol in eutopic endometrial cells of endometiosis, and its mechanism might be associated with combined action of inhibiting expression of SF-1,17-β-HSD 1 and up-regulating expression of COUP-TF,17-β-HSD 2.