Influence Of Clomiphene Citrate On Genomic Expression Of Endometrium And Its Machanism In Female Sprague-dawley Rats

Objectives:Clomiphene citrate,which was widely used in IVT-ET, has advantages of less cost, more conveniently taking and better effect on hyperstimulation. Although it was reported that clomiphene citrate has negtive effect on endometrial receptivity, inhibits cell proliferation and differentiation in endometrium, and leads to lower clinical pregnancy rate easily, clomiphene citrate was widely used in the classic short-and long-term stimulation and the better mini-stimulation protocol. However, the mechanism how clomiphene citrate influenced the proliferation and differentiation in endometrial cells is not clear yet. In this study, in order to research how CC take effect on endometrial receptivity, we plan to research genome expression changes in female rats’ endometrium after being medicated CC, discussing its mechanisms of CC to influence proliferation and differentiation in endometrial cells.Methods:In this study, we focused on female Sprague-Dawley rats, which were medicated orally through intragastric administration with CC, and according to duration of administration, these rats were randomly divided into6groups, Each of which group contains7rats medicated CC and2rat medicated normal saline. After administration, uteruses of the rats subsequently were stained with hematoxylin and eosin, in which endometrial thickness was observed based on histology. According to its result, we sellected5rats in6-day group and5rats in control group, extracted their total RNA in their uteruses and analysed expression of ralative genes with Rat estrogen receptor signaling PCR array from Qiagen. Considering the result of analyzation and the result of researching genes related to ESR signaling pathway on site of wikipathway, NCOAl expression was analyzed in endometrium of SD rats in different time of administration with Western Blotting, and then25rats were divided into5gourps, including group with CC, group with CC and E2, group with CC and HMG, group with CC, E2and HMG, and blank control (CC:lmg per lkg rat weight,; HMG:0.75IU per rat; E2:10mg per1kg rat weight), all of which have been medicated for6days, in whose endometrium c-myc expression was analyzed with Western Blot.Results:Endometrial thickness of administration groups decreased by CC, and the difference, between6-day group and control group, was greater than others, and NCOA1expression was down-regulated, which changed with oestrus cycles in different groups. Furthermore, c-myc has expression of high level, on which E2and HMG has no obvious effect.Conclusion:1Clomiphene Citrate can reduce endometrial thickness, and influence uterine receptivity;2Clomiphene Citrate up-regulated c-myc mRNA and protein levels, and downregulated NCOA1protein levels in the uterus;3Clomiphene Citrate had no significant effect on E2in the serum, but E2of the CC+E2+HMG group decreased significantly, and clomifene citrate has no significant effects on P, FSH and LH in serum.