Inhibitory effect on estrogen production and its influence on invasive ability of human endometrial cells of endometriosis by medicated serum of SLW

Objective: 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.Method: 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.Result: 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(P0.01).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(P0.01) while the expression of TIMP-1,2 protein could be increased obviously(P0.01).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 + E2 group 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.Conclusion: SLW could decrease the secretion of E2 so as to inhibit the invasion of the eutopic endometrial cells of endometriosis.