Zhou Yuanyuan, Li Hongping, Lin Chuanping, Mao Yuchan, Rao Jinpeng, Lou Yiyun, Yang Xinyun, Xu XiangRong, Jin Fan
Department of Reproductive Endocrinology, Women's Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Reproductive Medical Center, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
Reprod Toxicol. 2020 Dec;98:125-133. doi: 10.1016/j.reprotox.2020.09.005. Epub 2020 Sep 21.
Perfluorooctanoic acid (PFOA) has attracted widespread research attention as it is very stable, bioaccumulates, and causes reproductive toxicity. Data from several animal experiments and epidemiological studies indicate that female fertility may decline because of ovarian granulosa cell (GC) apoptosis as oocyte quality is positively associated with effective gap junctional intercellular communication (GJIC) between GCs. To the best of our knowledge, however, no previous trials have been conducted or reported on the effects of PFOA exposure on apoptosis induction in human GCs. Moreover, the roles of GJIC in GC survival and in the induction of apoptosis in GCs by PFOA remain unclear. To test this, we cultured human GCs in vitro and treated them with 0 μM, 0.3 μM, 3 μM, or 30 μM PFOA for 24 h. We also treated a human ovarian GC line (KGN) with various combinations of PFOA, retinoic acid (RA, 10 μM), and carbenoxolone disodium (CBX, 50 mM). Our findings showed that PFOA lowered human GC viability and increased apoptosis. The effects of CBX resemble those of PFOA. The combination of PFOA and CBX enhances the inhibition of GJIC by PFOA and promotes apoptosis. The effects of RA are the opposite to those of PFOA. The combination of RA and PFOA mitigates PFOA-induced GJIC inhibition and reduces apoptosis. The observed expression levels of apoptosis-related proteins were consistent with the aforementioned findings. Hence, our study demonstrated that PFOA may induce human ovarian GC apoptosis by inhibiting GJIC.
全氟辛酸（PFOA）因其非常稳定、具有生物累积性并会导致生殖毒性而受到广泛的研究关注。多项动物实验和流行病学研究的数据表明，由于卵巢颗粒细胞（GC）凋亡，女性生育能力可能会下降，因为卵母细胞质量与GC之间有效的间隙连接细胞间通讯（GJIC）呈正相关。然而，据我们所知，此前尚未进行过关于PFOA暴露对人GC凋亡诱导作用的试验或报道。此外，GJIC在GC存活以及PFOA诱导GC凋亡中的作用仍不清楚。为了验证这一点，我们在体外培养人GC，并分别用0 μM、0.3 μM、3 μM或30 μM的PFOA处理24小时。我们还用PFOA、视黄酸（RA，10 μM）和甘草次酸钠（CBX，50 mM）的不同组合处理人卵巢GC系（KGN）。我们的研究结果表明，PFOA降低了人GC的活力并增加了凋亡。CBX的作用与PFOA相似。PFOA和CBX的组合增强了PFOA对GJIC的抑制作用并促进了凋亡。RA的作用与PFOA相反。RA和PFOA的组合减轻了PFOA诱导的GJIC抑制并减少了凋亡。观察到的凋亡相关蛋白表达水平与上述结果一致。因此，我们的研究表明，PFOA可能通过抑制GJIC诱导人卵巢GC凋亡。
