CAJ | 학술논문

目的探讨NSP4与整联蛋白α2β1在轮状病毒致肝外胆管上皮细胞损伤中的作用.方法取Balb/c小鼠的肝外胆管进行肝外胆管上皮细胞培养至第三天,用激光共聚焦方法检测整联蛋白α2、β1亚基及轮状病毒NSP4.用siRNA干扰技术沉默培养至第三天的肝外胆管上皮细胞的整联蛋白α2后加入轮状病毒,用Real Time RT-PCR方法检测α2亚基及轮状病毒NSP4的mRNA),并进行统计学分析.结果用激光共聚焦方法检测到了整联蛋白α2、β1亚基存在于肝外胆管上皮细胞膜表面；轮状病毒NSP4从感染细胞向紧邻的非感染细胞进行扩散；siRNA干扰技术表明沉默整联蛋白α2后轮状病毒NSP4mRNA含量1±0.01较对照组0.56±0.09减低(P＜0.05).结论肝外胆管上皮细胞表面表达整联蛋白α2、β1且整联蛋白α2β1与轮状病毒NSP4相互作用并进一步导致肝外胆管上皮细胞损伤.
목적 탐토NSP4여정련단백α2β1재륜상병독치간외담관상피세포손상중적작용.방법 취Balb/c소서적간외담관진행간외담관상피세포배양지제삼천,용격광공취초방법검측정련단백α2、β1아기급륜상병독NSP4.용siRNA간우기술침묵배양지제삼천적간외담관상피세포적정련단백α2후가입륜상병독,용Real Time RT-PCR방법검측α2아기급륜상병독NSP4적mRNA),병진행통계학분석.결과 용격광공취초방법검측도료정련단백α2、β1아기존재우간외담관상피세포막표면；륜상병독NSP4종감염세포향긴린적비감염세포진행확산；siRNA간우기술표명침묵정련단백α2후륜상병독NSP4mRNA함량1±0.01교대조조0.56±0.09감저(P＜0.05).결론 간외담관상피세포표면표체정련단백α2、β1차정련단백α2β1여륜상병독NSP4상호작용병진일보도치간외담관상피세포손상.
Objective To investigate the role of rotavirus non structural protein 4 (NSP4) and integrin α2β1 in the damage of extrahepatic biliary epithclial cells.Methods Extrahepatic bilt duct of Balb/c mice was harvested for ex vivo culture of biliary epithelial cells (EHBE).The primary cultured epithelial cells werc fixed,Immunofluorescent (IF) assay was performed to detect integrin subunits a2,β1 and rotavirus nonstructural protein 4 (NSP4).Small interfering RNA targeting integrin α2 was transfected into EHBE before rotavirus infection.Real-time RT-PCR was used to measure the mRNAs of integrin α2 and NSP4.Results Integrin α2 and β1 were detected at the surface of EHBE.Furthermore,the expression of NSP4 was colocalized with integrin α2 on the surface of rotavirus-infected extrahepatic biliary epithelial cells,indicating that NSP4 released from rotavirus-infected EHBE hinded with the integrin α2 on the adjacent uninfected cells.Silencing of integrin α2 decreased the mRNA of rotavirus NSP4.Conclusions The integrin subunits α2,β1 are expressed and locate on the surface of primary cultured extrahepatic biliary epithelial cells.The integrin α2β1 and the rotavirus NSP4 play importnat roles in the injury of extrahepatic biliary epithelial cells.