Lu Yin, Luo Bin, Li Jing, Dai Jiayin
Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, People's Republic of China.
School of Life Science, Anhui University, Hefei, 230601, People's Republic of China.
Arch Toxicol. 2016 Apr;90(4):971-83. doi: 10.1007/s00204-015-1492-y. Epub 2015 Mar 6.
Perfluorooctanoic acid (PFOA) is correlated with male reproductive dysfunction in animals and humans, but the underlying mechanisms for this remain unknown. To explore the potential reproductive toxicity of PFOA, we studied blood-testis barrier (BTB) damage using in vivo and in vitro models. Male mice were gavage-administered PFOA (0-20 mg/kg/d) for 28 consecutive days, and breeding capacity and permeability of the Sertoli cell-based BTB were estimated. Primary Sertoli cells (SCs) were exposed to PFOA (0-500 μM) for 48 h, and transepithelial electrical resistance (TER) was assessed. Furthermore, BTB-associated protein expression, TNFα content, and phosphorylation and protein levels of the mitogen-activated protein kinase (MAPK) pathway were detected. An apparent decrease in mated and pregnant females per male mouse as well as litter weight was observed. Marked BTB damage was evidenced by increased red biotin fluorescence in the lumen tubular of the testes and the decrease in TER in SCs in vitro. The protein levels of claudin-11, connexin-43, N-cadherin, β-catenin, and occludin were significantly decreased in the testes and also in the SCs in vitro except for N-cadherin and β-catenin. TNFα content showed a dose-dependent increase in the testes and a dose- and time-dependent increase in the SCs, with the p-p38/p38 MAPK ratio also increasing in testes and SCs after PFOA exposure. Moreover, PFOA altered expressions of claudin-11, connexin-43, TNFα, and p-p38 MAPK were recovered 48 h after PFOA removal in the SCs. The SCs appeared to be target to PFOA, and the disruption of the BTB may be crucial to PFOA-induced reproductive dysfunction in mice.
全氟辛酸（PFOA）与动物和人类的雄性生殖功能障碍相关，但其潜在机制尚不清楚。为了探究PFOA的潜在生殖毒性，我们使用体内和体外模型研究了血睾屏障（BTB）损伤。雄性小鼠连续28天经口灌胃给予PFOA（0 - 20 mg/kg/d），评估基于支持细胞的BTB的生殖能力和通透性。原代支持细胞（SCs）暴露于PFOA（0 - 500 μM）48小时，评估跨上皮电阻（TER）。此外，检测了BTB相关蛋白表达、TNFα含量以及丝裂原活化蛋白激酶（MAPK）途径的磷酸化和蛋白水平。观察到每只雄性小鼠的交配和怀孕雌性数量以及窝仔重量明显减少。睾丸管腔中红色生物素荧光增加以及体外SCs中TER降低证明了明显的BTB损伤。除了N - 钙黏蛋白和β - 连环蛋白外，睾丸和体外SCs中紧密连接蛋白11、连接蛋白43、N - 钙黏蛋白、β - 连环蛋白和闭合蛋白的蛋白水平均显著降低。睾丸中TNFα含量呈剂量依赖性增加，SCs中呈剂量和时间依赖性增加，PFOA暴露后睾丸和SCs中p - p38/p38 MAPK比值也增加。此外，PFOA去除48小时后，SCs中紧密连接蛋白11、连接蛋白43、TNFα和p - p38 MAPK的表达改变得以恢复。SCs似乎是PFOA的靶细胞，BTB的破坏可能是PFOA诱导小鼠生殖功能障碍关键因素。
