Silencing of the rotavirus NSP4 protein decreases the incidence of biliary atresia in murine model.
Silencing of the rotavirus NSP4 protein decreases the incidence of biliary atresia in murine model.
复制标题
沉默轮状病毒 NSP4 蛋白可降低小鼠模型中胆道闭锁的发生率
DOI：
10.1371/journal.pone.0023655
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Chai C
中科院分区：
文献类型：
--
作者：
Feng J;Yang J;Zheng S;Qiu Y;Chai C
胆道闭锁是新生儿的常见病，可导致梗阻性黄疸和进行性肝纤维化。我们以前的研究表明，轮状病毒感染通过诱导核因子-kappaB升高和骨桥蛋白炎症通路的异常激活而在实验性胆道闭锁(BA)的发病机制中起始动作用。在轮状病毒感染的背景下，轮状病毒非结构蛋白4(NSP4)是重要的免疫原，病毒蛋白7(VP7)是轮状病毒成熟所必需的，病毒蛋白4(VP4)是病毒毒力的决定因素。本研究旨在阐明NSP4、VP7和VP4在实验性BA发病机制中的作用。用轮状病毒(Mmu18006)感染原代培养的肝外胆管上皮细胞。在体外和体内，针对NSP4、VP7或VP4的小干扰RNA在轮状病毒感染前被转染。我们分析了BA的发生率、病毒颗粒的形态变化、形态发生以及病毒的mRNA和蛋白的表达。体外实验表明，NSP4沉默可降低VP7和VP4水平，减少病毒颗粒，减少细胞病变效应。NSP4阳性细胞整合素亚单位α-2呈强阳性表达，沉默VP7或VP4可部分减轻上皮损伤。动物实验表明，NSP4沉默后，小鼠幼鼠的BA发生率低于VP7或VP4沉默后。然而，VP4沉默后33.3%的仔鼠(N = 6)出现BA，50%的N = 6出现胆道损伤。NSP4或VP4沉默后肝损伤减轻。NSP4后胆管内未检测到VP4和VP7。总之，NSP4沉默下调了VP7和VP4的表达，从而降低了BA的发生率。
Biliary atresia is a common disease in neonates which causes obstructive jaundice and progressive hepatic fibrosis. Our previous studies indicate that rotavirus infection is an initiator in the pathogenesis of experimental biliary atresia (BA) through the induction of increased nuclear factor-kappaB and abnormal activation of the osteopontin inflammation pathway. In the setting of rotavirus infection, rotavirus nonstructural protein 4 (NSP4) serves as an important immunogen, viral protein 7 (VP7) is necessary in rotavirus maturity and viral protein 4 (VP4) is a virulence determiner. The purpose of the current study is to clarify the roles of NSP4, VP7 and VP4 in the pathogenesis of experimental BA. Primary cultured extrahepatic biliary epithelia were infected with Rotavirus (mmu18006). Small interfering RNA targeting NSP4, VP7 or VP4 was transfected before rotavirus infection both in vitro and in vivo. We analyzed the incidence of BA, morphological change, morphogenesis of viral particles and viral mRNA and protein expression. The in vitro experiments showed NSP4 silencing decreased the levels of VP7 and VP4, reduced viral particles and decreased cytopathic effect. NSP4-positive cells had strongly positive expression of integrin subunit α2. Silencing of VP7 or VP4 partially decreased epithelial injury. Animal experiments indicated after NSP4 silencing, mouse pups had lower incidence of BA than after VP7 or VP4 silencing. However, 33.3% of VP4-silenced pups (N = 6) suffered BA and 50% of pups (N = 6) suffered biliary injury after VP7 silencing. Hepatic injury was decreased after NSP4 or VP4 silencing. Neither VP4 nor VP7 were detected in the biliary ducts after NSP4. All together, NSP4 silencing down-regulates VP7 and VP4, resulting in decreased incidence of BA.
登录
查看更多内容
影响因子：
2.4
作者：
Petersen, C;Biermanns, D;Mildenberger, H
通讯作者：
Mildenberger, H
影响因子：
3.8
作者：
Jafri, Mubeen;Donnelly, Bryan;Tiao, Greg
通讯作者：
Tiao, Greg
影响因子：
13.5
作者：
Tyler, KL;Sokol, RJ;Brown, WR
通讯作者：
Brown, WR
影响因子：
15.9
作者：
Shivakumar, P;Campbell, KM;Bezerra, JA
通讯作者：
Bezerra, JA
影响因子：
13.5
作者：
Harada, Kenichi;Sato, Yasunori;Nakanuma, Yasuni
通讯作者：
Nakanuma, Yasuni