Laboratory of Animal Pathology and Public Health, Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agriculture University, Beijing, China; Department of Pathobiology, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan.
Laboratory of Animal Pathology and Public Health, Key Laboratory of Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agriculture University, Beijing, China.
Virus Res. 2016 Feb 2;213:343-352. doi: 10.1016/j.virusres.2015.12.012. Epub 2015 Dec 24.
We examined the effect of hepatitis E virus (HEV) on the renal tissue pathogenesis, morphological damages and related molecular mechanisms following swine HEV suspension intraperitoneally inoculation in Mongolian gerbils. The microscopic and ultramicroscopic analyses of kidney tissue structure were carried out at different points after inoculation of HEV. The immunohistochemistry, real-time PCR and Western blot were performed to explore the molecular mechanisms associated with HEV presence in the renal tissues. Real-time PCR revealed that the copies of HEV RNA in the kidney were detected at 7 dpi, and peaked at 14 dpi at a concentration was 7.18 logs g(-1), with detection of HEV ORF2 antigen by immunohistochemistry. Hematoxylin and eosin (HE) staining showed pathological lesions including glomerular atrophy, degeneration, edema and necrosis of renal tubular epithelial cells and Mallory and Sirius red staining indicated the presence of collagen fibers and fibrosis in kidney tissues of inoculated gerbils. Ultrastructural studies of basal membrane of renal tubules demonstrated the rough and uneven with mitochondria swelling and vacuolation in the tissues of HEV inoculated animals. Similarly, significantly higher number of (TUNEL)-positive cells were seen in renal tubule tissues compared to control group. Moreover, immuno histochemical results indicated that significant increase expression of the B-cell lymphoma 2 (Bcl-2), Bcl-2 associated X protein (Bax), FAS and Caspase-3 in HEV inoculated Mongolian gerbils at each time points. Relative mRNA expression by real-time PCR revealed a significantly higher (P<0.05) mRNA level of BAX, Bcl-2 and caspase-3 transcription in HEV inoculated Mongolian gerbils. Our results demonstrates that activation of mitochondria and Caspase-3 protease might be induced the apoptosis which subsequently cause the necrosis and cell death of renal epithelial cells during acute phase of HEV infection in HEV inoculated Mongolian gerbils.
我们研究了戊型肝炎病毒（HEV）对蒙古沙土鼠肾组织发病机制、形态损伤及相关分子机制的影响，通过将 HEV 悬液腹腔接种于蒙古沙土鼠。在接种 HEV 后的不同时间点对肾组织结构进行了显微镜和超微结构分析。通过免疫组织化学、实时 PCR 和 Western blot 探索了与肾组织中 HEV 存在相关的分子机制。实时 PCR 显示，在 7dpi 时检测到肾组织中 HEV RNA 的拷贝数，并在 14dpi 时达到峰值，浓度为 7.18logs g(-1)，通过免疫组织化学检测到 HEV ORF2 抗原。苏木精和伊红（HE）染色显示包括肾小球萎缩、变性、肾小管上皮细胞水肿和坏死在内的病理损伤，马氏三色和天狼猩红染色显示接种沙土鼠肾组织中有胶原纤维和纤维化的存在。肾小管所基底膜的超微结构研究显示，粗糙不均匀，伴有线粒体肿胀和空泡化。同样，与对照组相比，在肾组织中观察到更多的（TUNEL）阳性细胞。此外，免疫组织化学结果表明，在 HEV 接种的蒙古沙土鼠中，B 细胞淋巴瘤 2（Bcl-2）、Bcl-2 相关 X 蛋白（Bax）、FAS 和 Caspase-3 的表达显著增加，在每个时间点。实时 PCR 的相对 mRNA 表达显示，HEV 接种的蒙古沙土鼠中 Bax、Bcl-2 和 caspase-3 转录的 mRNA 水平显著升高（P<0.05）。我们的结果表明，线粒体的激活和 Caspase-3 蛋白酶可能诱导细胞凋亡，进而导致急性 HEV 感染期间肾上皮细胞的坏死和细胞死亡。
