Silencing of the Rotavirus NSP4 Protein Decreases the Incidence of Biliary Atresia in Murine Model 英文参考文献.docVIP

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Silencing of the Rotavirus NSP4 Protein Decreases the Incidence of Biliary Atresia in Murine Model 英文参考文献.doc

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Silencing of the Rotavirus NSP4 Protein Decreases the Incidence of Biliary Atresia in Murine Model 英文参考文献

SilencingoftheRotavirusNSP4ProteinDecreasesthe IncidenceofBiliaryAtresiainMurineModel JiexiongFeng*,JixinYang,ShuaiyuZheng,YinrongQiu,ChengweiChai DepartmentofPediatricSurgery,TongjiHospital,TongjiMedicalCollege,HuazhongUniversityofScienceandTechnology,Wuhan,China Abstract 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.Inthesettingofrotavirusinfection,rotavirusnonstructuralprotein4(NSP4)servesasanimportantimmunogen, viralprotein7(VP7)isnecessaryinrotavirusmaturityandviralprotein4(VP4)isavirulencedeterminer.Thepurposeofthe current study is to clarify the roles of NSP4, VP7 and VP4 in the pathogenesis of experimental BA. Primary cultured extrahepaticbiliaryepitheliawereinfectedwithRotavirus(mmu18006).SmallinterferingRNAtargetingNSP4,VP7orVP4 wastransfectedbeforerotavirusinfectionbothinvitroandinvivo.WeanalyzedtheincidenceofBA,morphologicalchange, morphogenesisofviralparticlesandviralmRNAandproteinexpression.TheinvitroexperimentsshowedNSP4silencing decreased the levels of VP7 and VP4, reduced viral particles and decreased cytopathic effect. NSP4-positive cells had strongly positive expression of integrin subunit a2. 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%ofVP4-silencedpups(N=6)sufferedBAand50%ofpups(N=6)sufferedbiliaryinjuryafterVP7silencing. HepaticinjurywasdecreasedafterNSP4orVP4silencing.NeitherVP4norVP7weredetectedinthebiliaryductsafterNSP4. Alltogether,NSP4silencingdown-regulatesVP7andVP4,resultingindecreasedincidenceofBA. Citation:FengJ,YangJ,ZhengS,QiuY,ChaiC(2011)SilencingoftheRotavirusNSP4Pr