戊型肝炎病毒与UBR2相互作用的初步研究

A preliminary study on the interaction between hepatitis E virus and UBR2

戊型肝炎病毒(hepatitis E virus, HEV)是一种经肠道传播、严重危害人类健康的病毒性肝炎病原体。HEV不仅可以在肝脏中复制,还可以在肝外组织复制并导致组织损伤,如HEV可在男性生殖系统中复制并导致睾丸损伤,但其发病机制尚不明确。UBR2在生殖系统高表达,是男性减数分裂和精子形成的关键,但其与HEV的相互作用尚不清楚。本文以探究HEV与UBR2之间的相互作用关系为目的,通过构建真核表达载体、转染细胞,利用Western blot技术验证UBR2能否成功过表达。同时,在细胞中加入HEV病毒,并利用Western blot技术探究HEV对UBR2的影响。另外,在HEV感染的新西兰大白兔睾丸组织中,利用免疫荧光共定位技术观察HEV与UBR2的相互作用关系。上述实验成功构建了pEGFP-UBR2质粒,并在A549细胞和Huh 7.5.1细胞中过表达,发现在A549细胞系中,HEV对UBR2有抑制作用;在感染HEV的睾丸组织中发现高表达的UBR2与HEV ORF2衣壳蛋白能够共定位,提示了两者可能存在相互作用。这为深入研究HEV与UBR2的相互作用,以及HEV导致生殖系统损伤的致病机制奠定了基础。

Hepatitis E virus(HEV) is a main pathogen of viral hepatitis, which is transmitted by fecesoral route. HEV replicates in liver and other extra-hepatic tissues. For example, HEV replicates in the male reproductive system and causes testicular damage. However, the pathogenesis still unclear. UBR2 highly expresses in the male reproductive system, which is critical to sperm meiosis and development, but its relationship with HEV infection is rarely reported. In this study, UBR2 gene was introduced into eukaryotic expression vector pEGFP-N1 and transfected A549 cells or Huh 7.5.1 cells to explore the interaction between HEV and UBR2. The results of Western blot analysis showed that UBR2 was successfully over-expressed in both A549 and Huh 7.5.1 cells. To assess the association between HEV and UBR2, cells were pre-transfected with UBR2 then infected with HEV. Western blot results indicated that HEV significantly inhibited the expression of UBR2. In addition, the co-location of HEV and UBR2 strongly suggested that HEV interacted with UBR2 in the testis of HEV-infected rabbits. Our results indicated that HEV interacted with UBR2 in vitro and in vivo, which will highlight the knowledge of male reproductive system damages caused by HEV.