This study was undertaken to evaluate the role of progesterone (P4) in modulation of the expression profile of adhesion‐related molecules in uterine epithelial cells (UECs) and in vitro blastocyst production in buffalo. UECs were isolated from slaughterhouse‐derived uteri by enzymatic treatment, and cells were characterized by immunocytochemistry (ICC) and PCR assays. The well‐characterized UECs were exposed to different concentrations of P4 (0, 0.314, 3.14 and 6.28 ng/ml) along with the basal level of oestradiol for 6 days. Thereafter, the relative mRNA expression of different biomolecules such as mucin 1 (MUC1), osteopontin, integrin alpha (α3, α6 and αV) and beta (β1 and β3) subunits, progesterone receptor (PR) and oestrogen receptor, was evaluated. Further, day 2 post‐insemination embryos were cultured in mSOF supplemented with or without P4. UECs were found positive for cytokeratin expression and negative for vimentin expression. Progesterone treatment significantly enhanced the mRNA expression of most of the transcripts compared with the control group, and correspondingly, the immunofluorescence depicted higher protein expression of all these molecules. Further, the long‐term exposure of UECs to P4 downregulated the expression of PR and, concomitantly, MUC1. Progesterone supplementation to embryo culture medium significantly (p  < .05) improved the blastocyst rate. The study demonstrates the role of P4 hormone in modulation of the expression of early implantation‐related biomolecules in uterine epithelial cells; hence, adequate level of steroids is crucial for normal embryo development and its implantation.

中文翻译：

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孕酮调节子宫上皮细胞中的黏附分子和水牛的体外胚胎产生。

这项研究旨在评估孕酮（P4）在调节子宫上皮细胞（UEC）中粘附相关分子的表达谱以及在水牛中体外胚泡产生中的作用。通过酶处理从屠宰场衍生的子宫中分离出UEC，并通过免疫细胞化学（ICC）和PCR分析对细胞进行表征。表征良好的UEC暴露于不同浓度的P4（0、0.314、3.14和6.28 ng / ml）以及基础剂量的雌二醇6天。此后，评估了诸如粘蛋白1（MUC1），骨桥蛋白，整联蛋白α（α3，α6和αV）和β（β1和β3）亚基，孕激素受体（PR）和雌激素受体等不同生物分子的相对mRNA表达。此外，将第2天的受精后胚胎在添加或不添加P4的mSOF中培养。发现UECs对细胞角蛋白表达呈阳性，而对波形蛋白表达呈阴性。与对照组相比，孕酮处理显着提高了大多数转录本的mRNA表达，相应地，免疫荧光显示所有这些分子的蛋白质表达均较高。此外，UEC长期暴露于P4下调了PR的表达，并因此下调了MUC1的表达。黄体酮对胚胎培养基的显着补充（UEC长期暴露于P4下调了PR的表达，并同时下调了MUC1的表达。黄体酮对胚胎培养基的显着补充（UEC长期暴露于P4下调了PR的表达，并同时下调了MUC1的表达。孕激素显着补充胚胎培养基（p  <.05）提高了胚泡率。这项研究证明了P4激素在调节子宫上皮细胞中与早期植入有关的生物分子表达中的作用。因此，适当水平的类固醇对于正常的胚胎发育及其植入至关重要。