Wang Wei, Ge Liang, Zhang Li-Li, Wang Li-Rong, Lu Yong-Yan, Gou Li, Gou Rui-Qiang, Xu Tong-Yu, Ma Xiao-Ling, Zhang Xue-Hong
The Reproductive Medicine Center of the First Hospital of Lanzhou University, Key Laboratory for Reproductive Medicine and Embryo, Lanzhou, Gansu, China.
Department of Anesthesiology, Gansu Province Maternity and Child-care Hospital, Lanzhou, Gansu, China.
Kaohsiung J Med Sci. 2023 May;39(5):468-477. doi: 10.1002/kjm2.12672. Epub 2023 Mar 13.
Human chorionic gonadotropin (hCG) might affect endometrial receptivity, exerting integral roles in embryo implantation. This study explored the action of hCG in endometrial receptivity via the miR-126-3p/PIK3R2/PI3K/Akt/eNOS axis. The embryo implantation dysfunction (EID) mouse models were established by administrating mifepristone and human endometrial epithelial cells (EECs) were used for in vivo experiments, both followed by hCG treatment. Expression level of CD105 and protein levels of cadherin CD144 and CD146 in mice were determined by immunohistochemistry and Western blot. The levels of miR-126-3p and PIK3R2 mRNA and PIK3R2, p-PI3K p85 α, PI3K p110 α, p-Akt, Akt, p-eNOS, and eNOS protein levels were measured. Cell proliferation was evaluated by CCK-8 and EdU assays. The binding sites of miR-126-3p and PIK3R2 were predicted and verified. hCG-treated EECs were further transfected with miR-126-inhibitor for functional rescue experiments. hCG ameliorated endometrial receptivity in EID mice. Moreover, hCG promoted miR-126-3p and suppressed PIK3R2 in EID mice and EECs. miR-126-3p targeted PIK3R2. EEC proliferation was enhanced after hCG treatment but inhibited by miR-126-3p downregulation. Both in vivo and in vitro experiments validated that hCG activated the PI3K/Akt/eNOS pathway through the miR-126-3p/PIK3R2 axis. Collectively, hCG improves endometrial receptivity by activating the PI3K/Akt/eNOS pathway via regulating miR-126-3p/PIK3R2.
人绒毛膜促性腺激素（hCG）可能会影响子宫内膜容受性，在胚胎着床过程中发挥重要作用。本研究通过miR-126-3p/PIK3R2/PI3K/Akt/eNOS轴探讨了hCG在子宫内膜容受性中的作用机制。通过给予米非司酮建立胚胎着床功能障碍（EID）小鼠模型，并将人子宫内膜上皮细胞（EECs）用于体内实验，随后进行hCG处理。通过免疫组织化学和蛋白质印迹法检测小鼠中CD105的表达水平以及钙黏蛋白CD144和CD146的蛋白质水平。检测miR-126-3p和PIK3R2 mRNA水平以及PIK3R2、p-PI3K p85α、PI3K p110α、p-Akt、Akt、p-eNOS和eNOS蛋白质水平。通过CCK-8和EdU实验评估细胞增殖。预测并验证了miR-126-3p与PIK3R2的结合位点。用miR-126抑制剂进一步转染hCG处理的EECs进行功能挽救实验。hCG改善了EID小鼠的子宫内膜容受性。此外，hCG促进了EID小鼠和EECs中miR-126-3p的表达并抑制了PIK3R2的表达。miR-126-3p靶向PIK3R2。hCG处理后EEC增殖增强，但miR-126-3p下调则抑制了增殖。体内和体外实验均证实，hCG通过miR-126-3p/PIK3R2轴激活PI3K/Akt/eNOS通路。总之，hCG通过调节miR-126-3p/PIK3R2激活PI3K/Akt/eNOS通路，从而改善子宫内膜容受性。
