FATP5基因沉默抑制油酸诱导的HepG2细胞脂肪化及炎症作用  

Effect of FATP5 gene silencing on fatty hepatic cell inflammation

作　　者：王天琦 孙文龙 杨馥源 李敬达 WANG Tian-qi;SUN Wen-long;YANG Fu-yuan;LI Jing-da(College of Agriculture,Yangtze University,Jingzhou Hubei 434000,China;College of Life Sciences,Shandong University of Technology,Zibo Shandong 255000,China;Medical College Yangtze University,Jingzhou Hubei 434000,China;College of Life Science,Yangtze University,Jingzhou Hubei 434000,China)

机构地区：[1]长江大学农学院,湖北荆州434000 [2]山东理工大学生命科学学院,山东淄博255000 [3]长江大学医学院,湖北荆州434000 [4]长江大学生命科学学院,湖北荆州434000

出　　处：《中国药理学通报》2023年第4期692-699,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学青年科学基金项目(No 82004020,81903878)。

摘　　要：目的 明确FATP5作为脂肪酸转运蛋白家族成员,其基因沉默对肝脏细胞脂肪化及炎症作用的影响,并初步探究该影响可能的机制。方法 设计合成5条shRNA序列,并借助siCHECK^(TM)系统筛选得到FATP5有效干扰序列。将有效干扰序列包装成慢病毒后,通过细胞感染和嘌呤霉素筛选获得FATP5基因沉默HepG2细胞株。使用Western blot方法检测FATP5基因沉默的效率。之后,利用检测试剂盒、Western blot、核质分离及报告基因方法分别分析FATP5基因沉默细胞中油酸诱导ROS和MDA生成、TNF-α和IL-6蛋白表达与分泌,以及NF-κB活化的变化情况。结果 基因沉默细胞中FATP5蛋白表达水平下降90%,并且脂质蓄积作用也被显著抑制。此外,FATP5基因沉默可减少油酸诱导的ROS及MDA生成,同时还可降低NF-κB的转录活性,并下调其下游炎症因子TNF-α及IL-6的蛋白表达和分泌。结论FATP5基因沉默可减轻脂肪化肝脏细胞的炎症作用,其机制可能与抑制氧化应激和脂质过氧化作用有关。Aim To determine the effect of FATP5 gene silencing on fatty hepatic cell inflammation and to explore its possible mechanism.Methods Five shRNA sequences were designed and synthesized.The efficient FATP5-shRNA was screened by the siCHECK ^(TM) system.After preparing the FATP5-shRNA lentivirus,the FATP5 gene silence hepatic cell lines was obtained by HepG2 cell infection and puromycin screening.The FATP5 silencing efficiency was detected by Western blot.Then the oleic acid induced ROS and MDA generation,TNF-αand IL-6 protein expression and secretion,and NF-κB activation in FATP5 gene silence cells were analyzed by the detection kit,Western blot,nucleo-plasmic separation and reporter gene system.Results In the gene silence cells,FATP5 protein expression was reduced by 90%and the lipid accumulation was also significantly inhibited.Moreover,the FATP5 knockdown could reduce the oleic acid induced ROS and MDA generation,and suppress the NF-κB activation,thereby inhibiting the protein expression and secretion of TNF-αand IL-6.Conclusions FATP5 gene silence inhibits fatty hepatic cell inflammation,and its mechanism may be related to the inhibition of oxidative stress and lipid peroxidation.

关 键 词：FATP5受体 shRNA基因干扰 炎症反应 非酒精性脂肪肝 NF-κB TNF-α 

分 类 号：R342.2[医药卫生—基础医学] R344.7R364.5R392.11R575.5