过表达MORC2通过下调p53含量抑制L02肝细胞脂肪变性.docVIP

DOI：10.16016/j.1000-5404.201502051 过表达MORC2p53含量抑制L02肝细胞脂肪变性 王 涛，雷增杰，王 斌，李 青，王春华，刘 琴，魏艳玲，王 军，陈东风 （400042 重庆，第三军医大学大坪医院野战外科研究所消化内科） [摘要] 目的 研究过表达MORC2对人L02肝细胞脂肪变性程度的影响及可能机制。方法 利用慢病毒转染技术在L02肝细胞中过表达MORC2，qRT-PCR及Western blot鉴定过表达效果后，对L02肝细胞进行脂肪变性诱导，测定细胞甘油三酯（triglyceride，TG）含量并比较过表达前后L02肝MORC2前后的基因表达谱进行检测与Pathway分析，筛选过表达MORC2时发生变化的通路并选择对脂代谢有调控作用的p53进行研究； Western blot检测在MORC2对p53蛋白含量的影响；在过表达MORC2的基础上对L02肝细胞p53，qRT-PCR及Western blot鉴定p53过表达效果后，通过TG测定观察过表达p53对过表达MORC2改变的L02肝细胞脂肪蓄积程度产生的影响。 结果 qRT-PCR及Western blot证明过表达MORC2成功；过表达MORC2的L02肝细胞在脂肪变性诱导后TG含量较MORC2正常表达的L02肝细胞明显降低（P0.01）；基因表达谱芯片及Pathway分析显示脂代谢调控相关的p53通路在过表达MORC2后发生变化；Western blot显示过表达MORC2后，L02肝细胞中p53蛋白水平在脂肪变性诱导前后均降低；qRT-PCR及Western blot验证p53过表达成功，且过表达p53可部分解除MORC2对L02肝细胞脂肪变性的缓解作用。结论 过表达MORC2p53从而缓解肝细胞脂肪变性。 [关键词] 非酒精性脂肪性肝病；脂肪变性；L02肝细胞；MORC2；p53 [中图法分类号] R322.47；R329.25；R363.2 [文献标志码] A Overexpression of MORC2 attenuates steatosis of L02 hepatocytes by downregulating p53 abundance Wang Tao, Lei Zengjie, Wang Bin, Li Qing, Wang Chunhua, Liu Qin, Wei Yanling, Wang Jun, Chen Dongfeng (Department of Gastroenterology, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042) [Abstract] Objective To investigate the effect of MORC2 overexpression on steatosis of immortalized hepatocyte cell line L02 and the potential mechanisms. Methods ?qRT-PCR and Western Blot were applied to examine overexpression of MORC2 in lentivirus-transfected L02 hepatocytes. Intracellular triglyceride (TG) was measured to evaluate cellular steatosis. Microarray chip was performed to analyze signaling pathways that were modulated in MORC2-expressing L02 cells. We further examined overexpression of p53 protein level by western blot in MORC2-overexpressing L02 cells before and after steatosis induction. Results Both qRT-PCR and Western Blot data validated robust increases in mRNA and protein levels of MORC2 in lentivirus-transfected L02 hepatocytes. Steatosis level was significant