Department of Veterinary Medicine, College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang 524088, China.
Department of Animal Science, College of Coastal Agricultural Sciences, Guangdong Ocean University, Zhanjiang 524088, China.
Viruses. 2023 May 29;15(6):1272. doi: 10.3390/v15061272.
The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) disrupts the blood-testis barrier (BTB), resulting in alterations in spermatogenesis. However, whether BTB-related proteins (such as ZO-1, claudin11, N-cadherin, and CX43) are targeted by SARS-CoV-2 remains to be clarified. BTB is a physical barrier between the blood vessels and the seminiferous tubules of the animal testis, and it is one of the tightest blood-tissue barriers in the mammalian body. In this study, we investigated the effects of viral proteins, via ectopic expression of individual viral proteins, on BTB-related proteins, the secretion of immune factors, and the formation and degradation of autophagosomes in human primary Sertoli cells. Our study demonstrated that ectopic expression of viral E (envelope protein) and M (membrane protein) induced the expressions of ZO-1 and claudin11, promoted the formation of autophagosomes, and inhibited autophagy flux. S (spike protein) reduced the expression of ZO-1, N-cadherin, and CX43, induced the expression of claudin11, and inhibited the formation and degradation of autophagosomes. N (nucleocapsid protein) reduced the expression of ZO-1, claudin11, and N-cadherin. All the structural proteins (SPs) E, M, N, and S increased the expression of the FasL gene, and the E protein promoted the expression and secretion of FasL and TGF-β proteins and the expression of IL-1. Blockage of autophagy by specific inhibitors resulted in the suppression of BTB-related proteins by the SPs. Our results indicated that SARS-CoV-2 SPs (E, M, and S) regulate BTB-related proteins through autophagy.
严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）破坏血睾屏障（BTB），导致精子发生改变。然而，SARS-CoV-2 是否靶向 BTB 相关蛋白（如 ZO-1、claudin11、N-钙黏蛋白和 CX43）仍有待阐明。BTB 是动物睾丸血管和生精小管之间的物理屏障，是哺乳动物体内最紧密的血组织屏障之一。在这项研究中，我们通过异位表达单个病毒蛋白，研究了病毒蛋白对 BTB 相关蛋白、免疫因子分泌以及自噬体形成和降解的影响在人原代支持细胞中。我们的研究表明，病毒 E（包膜蛋白）和 M（膜蛋白）的异位表达诱导了 ZO-1 和 claudin11 的表达，促进了自噬体的形成，并抑制了自噬流。S（刺突蛋白）降低了 ZO-1、N-钙黏蛋白和 CX43 的表达，诱导了 claudin11 的表达，并抑制了自噬体的形成和降解。N（核衣壳蛋白）降低了 ZO-1、claudin11 和 N-钙黏蛋白的表达。所有结构蛋白（SPs）E、M、N 和 S 均增加了 FasL 基因的表达，E 蛋白促进了 FasL 和 TGF-β 蛋白的表达和分泌以及 IL-1 的表达。特异性抑制剂阻断自噬导致 SPs 抑制 BTB 相关蛋白。我们的结果表明，SARS-CoV-2 SPs（E、M 和 S）通过自噬调节 BTB 相关蛋白。
