Li Ao, Wang Ying, Dong Wei, Xu Xiaoyu
College of Chemistry and Bioengineering, Chongqing University of Technology, Chongqing 400050, China.
Zhongguo Zhong Yao Za Zhi. 2010 Jun;35(12):1607-11.
To study the inhibitory effect of medicated serum of SLW on estrogen production and to approach on the key mechanism of SLW in inhibiting the invasion ability of endometrial cells of endometriosis.
First, the model of eutopic primary cultured endometrial cells of endometriosis and hysteromyoma in vitro was successfully established. Taking that of 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, the activities of matrix metalloproteinase-2, 9 (MMP-2, 9) were detected by gelatinase zymography assay, and the expression of tissue inhibitor of metalloproteinase-1, 2 (TIMP-1, 2) protein was observed by immunofluorescence. After the optimal time for SLW to inhibit invasion ability of endometrial cells was identified based on time-effect relationship, another endometrial 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 + middle dose of SLW serum + E2 group, eutopic endometrium of endometriosis + anastrozole serum group , and eutopic endometrium of endometriosis + E2 group. The activities of MMP-2, 9 and the expression of TIMP-1, 2 protein were detected according to the optimal time point.
The secretion level of E2 of eutopic endometrium in endometriosis could be decreased by SLW, which showed the dependence of time and concentration. The result of gelatinase zymography assay and immunofluorescence respectively showed that along with the time the activities of MMP-2, 9 of eutopic endometrial cells of endometriosis were significantly higher than those of hysteromyoma at the same time point (P < 0.01). After 48 hours, with the treatment of middle dose of SLW serum, the activities of MMP-2, 9 of eutopic endometrial cells of endometriosis could be decreased (P < 0.01) while the expression of TIMP-1, 2 protein could be increased obviously (P < 0.01). The malignant invasion ability improved by SLW of eutopic endometrial cells of endometriosis was partly recruited by add-back E2 treatment. There was no significant difference in the activity of MMP-2 and the expression of TIMP-1, 2 protein between eutopic endometrium of endometriosis + middle dose of SLW serum + E2group and untreated group. The behavior of invasion of endometrial cells of endometriosis could be deteriorated treated by E2 as contrast to anastrozole, a specific aromatase inhibitor.
SLW could decrease the secretion of E2 so as to inhibit the invasion of the eutopic endometrial cells of endometriosis.
研究四棱草根含药血清对雌激素生成的抑制作用，探讨四棱草根抑制子宫内膜异位症在位内膜细胞侵袭能力的关键机制。
首先成功建立子宫内膜异位症和子宫肌瘤在位原代培养子宫内膜细胞的体外模型。以子宫肌瘤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、低剂量四棱草根血清处理不同时间点后培养基上清中在位内膜细胞的E2分泌水平，用明胶酶谱法检测基质金属蛋白酶-2、9（MMP-2、9）的活性，用免疫荧光法观察金属蛋白酶组织抑制剂-1、2（TIMP-1、2）蛋白的表达。根据时间效应关系确定四棱草根抑制在位内膜细胞侵袭能力的最佳时间后，将另一批在位内膜细胞分为6组：子宫肌瘤内膜组、子宫内膜异位症在位内膜组、子宫内膜异位症在位内膜+中剂量四棱草根血清组、子宫内膜异位症在位内膜+中剂量四棱草根血清+E2组、子宫内膜异位症在位内膜+阿那曲唑血清组、子宫内膜异位症在位内膜+E2组。按最佳时间点检测MMP-2、9的活性及TIMP-1、2蛋白的表达。
四棱草根可降低子宫内膜异位症在位内膜的E2分泌水平，呈时间和浓度依赖性。明胶酶谱法及免疫荧光结果分别显示，随着时间推移，子宫内膜异位症在位内膜细胞的MMP-2、9活性在同一时间点均显著高于子宫肌瘤内膜细胞（P<0.01）。48小时后，中剂量四棱草根血清处理可降低子宫内膜异位症在位内膜细胞的MMP-2、9活性（P<0.01），同时明显增加TIMP-1、2蛋白的表达（P<0.01）。E2回补处理部分逆转了四棱草根对子宫内膜异位症在位内膜细胞恶性侵袭能力的改善作用。子宫内膜异位症在位内膜+中剂量四棱草根血清+E2组与未处理组之间MMP-2活性及TIMP-1、2蛋白表达无显著差异。与特异性芳香化酶抑制剂阿那曲唑相比，E2处理可恶化子宫内膜异位症在位内膜细胞的侵袭行为。
四棱草根可降低E2分泌，从而抑制子宫内膜异位症在位内膜细胞的侵袭。
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