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缺血再灌注对小鼠肝细胞自噬与干扰素调节因子1表达的影响 被引量:4

Effect of ischemia-reperfusion on the autophagy and interferon regulatory factor 1 expression in mouse livers
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摘要 目的 探讨缺血再灌注对小鼠肝细胞自噬与干扰素调节因子1(IRF-1)表达的影响,阐述其潜在的分子机制.方法 建立小鼠肝缺血再灌注损伤模型,检测再灌注2、6、12和24 h后各组与假手术组小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)的变化,同时在光镜下观察肝组织病理学改变,透射电镜下观察肝细胞内自噬体变化.分别利用免疫荧光及TUNEL法检测肝细胞LC3表达及细胞凋亡情况,RT-PCR检测肝组织IRF-1 mRNA表达,蛋白印迹检测IRF-1、Beclin1和LC3蛋白水平.结果 与假手术组相比,缺血再灌注组在各时间点ALT和AST水平均有所增高,于再灌注12 h均达到高峰;病理学检查可见肝细胞肿胀、肝窦变窄、嗜中性粒细胞浸润和灶状坏死等变化.再灌注12h时肝细胞内自噬体明显增多,同时凋亡细胞显著增加;肝组织中IRF-1、Beclin1和LC3的表达增加.结论 小鼠肝缺血再灌注可激活自噬导致肝细胞损伤,其可能与促进IRF-1表达有关. Objective To explore the effect of ischemia-reperfusion (IR) on autophagy and interferon regulatory factor 1 (IRF-1) expression in mouse liver.Methods Hepatic IR injury (IRI) model was established in C57BL/6 mice,and the serum AST and ALT levels were tested in all groups (Sham and IR) at 2 h,6 h,12 h and 24 h,respectively.Histopathological feature of IRI was observed,and autophagosomes changes were observed by transmission electron microscope.LC3 expression was detected by immunofluorescence and cell apoptosis was detected by TUNEL assay.IRF-1 mRNA and IRF-1,LC3 and Beclin1 protein were detected by RT-PCR and western blot,respectively.Results As the reperfusion progressed,serum AST and ALT levels in the mice were increased and peaked at 12 h.Hepatocyte swelling,hepatic sinusoids narrowing,inflammatory cell infiltration and hepatocyte necrosis were observed in the livers from IR group.Compared with Sham group,there was a significant increase on the number of autophagosomes in IR group.In addition,apoptotic cells were also significantly increased.The expression of IRF-1,Beclin1,and LC3 levels were up-regulated in IR group compared to Sham group.Conclusion IR could activate autophagy to cause hepatocyte damage,which may be related to the up-regulation of IRF-1.
出处 《中华肝胆外科杂志》 CAS CSCD 北大核心 2016年第2期111-115,共5页 Chinese Journal of Hepatobiliary Surgery
基金 国家高技术研究发展计划(863),国家自然科学基金,国家临床重点专科建设项目(器官移植),卫生公益性行业科研专项,National High Technology Research and Development Program (863) of China,National Natural Science Foundation of China,National key specialty construction of clinical projects (organtransplantation),Special Fund for Health Research in the Public Interest of China
关键词 缺血再灌注 肝细胞 自噬 干扰素调节因子1 Ischemia-reperfusion Hepatocytes Autophagy Interferon regulatory factor 1
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