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孕酮调节水牛子宫上皮细胞中的黏附分子和体外胚胎生产。

Progesterone modulates adhesion molecules in uterine epithelial cells and in vitro embryo production in buffalo.

机构信息

Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, India.

Department of Veterinary Physiology & Biochemistry, Faculty of Veterinary and Animal Sciences, RGSC, BHU, Barkachha, India.

出版信息

Reprod Domest Anim. 2020 Jul;55(7):833-843. doi: 10.1111/rda.13691. Epub 2020 May 18.

Abstract

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.

摘要

本研究旨在评估孕酮(P4)在调节水牛子宫上皮细胞(UEC)中粘附相关分子表达谱和体外囊胚产生中的作用。通过酶处理从屠宰场来源的子宫中分离出 UEC,并通过免疫细胞化学(ICC)和 PCR 检测法对细胞进行特征描述。将这些特征明确的 UEC 暴露于不同浓度的 P4(0、0.314、3.14 和 6.28ng/ml)以及基础水平的雌二醇 6 天。此后,评估了不同生物分子(如粘蛋白 1(MUC1)、骨桥蛋白、整合素α(α3、α6 和αV)和β(β1 和β3)亚基、孕激素受体(PR)和雌激素受体)的相对 mRNA 表达。进一步,在添加或不添加 P4 的 mSOF 中培养第 2 天授精的胚胎。UEC 对细胞角蛋白表达呈阳性,对波形蛋白表达呈阴性。与对照组相比,孕酮处理显著增强了大多数转录物的 mRNA 表达,相应地,免疫荧光显示所有这些分子的蛋白表达更高。此外,UEC 长期暴露于 P4 下调了 PR 的表达,同时,MUC1 的表达也下调了。胚胎培养液中添加孕酮显著(p<0.05)提高了囊胚率。该研究证明了 P4 激素在调节子宫上皮细胞中早期着床相关生物分子表达中的作用;因此,适当水平的类固醇对于正常胚胎发育和着床至关重要。

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