College of Animal Science, Yangtze University, Jingzhou, 434025, Hubei, China; College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
College of Animal Science and Technology, Northeast Agricultural University, Harbin, 150030, Heilongjiang, China.
Theriogenology. 2022 Dec;194:64-74. doi: 10.1016/j.theriogenology.2022.09.027. Epub 2022 Sep 29.
Heat shock protein 90 (HSP90), as a molecular chaperone, regulates hundreds of protein clients under both physiological and stress conditions in eukaryotic cells. However, the functional role of HSP90 in mammalian male reproduction remains largely unknown. Here, we aimed to investigate the function and effect of HSP90AA1 on the basic and reproductive function of pig immature Sertoli cells (iSCs). We first confirmed that the transfection of pBI-CMV3-HSP90AA1 vector into porcine iSCs for 24 h significantly increased mRNA and protein levels of HSP90AA. Moreover, HSP90AA1 over-expression significantly increased cell viability and the PLK2 mRNA abundance, promoted lactate production via elevating the LDHA activity, and inhibited the secretion of anti-Mullerian hormone and estradiol. In comparison, HSP90AA inhibition by allylamino-17-demethoxygeldanamycin (17-AAG) (2 μM) treatment of pig iSCs for 36 h had a totally contrasting effect, i.e. significantly reduced cell viability, promoted cell apoptosis via modulating expression of genes related to cell cycle and apoptosis (CCNB1, CCN1, PLK2, PTMA, YBX3 and CASP3), suppressed lactate production via dropping LDHA activity, but increased the secretion of anti-Mullerian hormone and estradiol. Taken together, our findings demonstrated that HSP90AA1 could regulate positively cell viability and lactate production, but negatively the secretion of reproductive hormones (anti-Mullerian hormone and estradiol). However, the detailed molecular mechanism of HSP90AA1 remains to be investigated.
热休克蛋白 90（HSP90）作为一种分子伴侣，在真核细胞的生理和应激条件下调节数百种蛋白质客户。然而，HSP90 在哺乳动物雄性生殖中的功能作用在很大程度上仍然未知。在这里，我们旨在研究 HSP90AA1 对猪未成熟支持细胞（iSCs）基本和生殖功能的作用。我们首先证实，将 pBI-CMV3-HSP90AA1 载体转染到猪 iSCs 中 24 小时，可显著增加 HSP90AA1 的 mRNA 和蛋白水平。此外，HSP90AA1 的过表达显著增加了细胞活力和 PLK2 mRNA 的丰度，通过提高 LDHA 活性促进了乳酸的产生，并抑制了抗苗勒管激素和雌二醇的分泌。相比之下，HSP90AA1 的抑制剂 17-AAG（2 μM）处理猪 iSCs 36 小时则产生了完全相反的效果，即显著降低了细胞活力，通过调节与细胞周期和凋亡相关的基因（CCNB1、CCN1、PLK2、PTMA、YBX3 和 CASP3）的表达，促进了细胞凋亡，抑制了乳酸的产生通过降低 LDHA 活性，但增加了抗苗勒管激素和雌二醇的分泌。总之，我们的研究结果表明，HSP90AA1 可以正向调节细胞活力和乳酸的产生，但负向调节生殖激素（抗苗勒管激素和雌二醇）的分泌。然而，HSP90AA1 的详细分子机制仍有待研究。
