非酒精性脂肪变性肝细胞模型中ATF4基因的表达.doc

非酒精性脂肪变性肝细胞模型中ATF4基因的表达及其意义 王万东，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 研究内质网应激相关蛋白转录活化因子4（activating transcription factor 4, ATF4）基因在油酸诱导脂肪变性的肝细胞模型中的表达变化及其意义。方法 以常规培养的L02肝细胞为对照，采用油酸诱导L02肝细胞脂肪变性，分别于0、24、48、72 h采用RT-PCR与Western blot方法动态监测ATF4基因的表达变化。结果 随着时间的增长，肝细胞脂肪变性程度加剧，肝细胞中ATF4 mRNA表达量0 、24、48、72 h分别为1.01±0.12、1.87±0.24、2.01±0.13、1.79±0.19，48 h时达到峰值后开始缓慢下降；ATF4蛋白表达量0 、24、48、72 h分别为0.31±0.16、0.57±0.14、0.91±0.20、0.89±0.17，直至72 h时仍维持在高水平状态，各试验组（24、48、72 h组）与空白对照组（0 h组）比较差异有显著性（P0.05）。结论 脂肪变性肝细胞中ATF4表达增强，说明肝细胞脂肪变性可能引起内质网发生应激，其可能与细胞非酒精性脂肪性肝病关系密切。 [关键词] 非酒精性脂肪肝; 细胞, 培养的; 转录活化因子4 [中图法分类号] [文献标志码] A Expression and significance of activating transcription factor 4 in oleic acid-induced nonalcoholic fatty liver disease in hepatocyte steatosis in vitromodels induced by Wang Wandong, Chen Dongfeng (Department of Gastroenterology, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China) [Abstract] Objective To investigate the expression and significance of activating transcription factor 4 (ATF4) in the hepatocyte steatosis models induced by oleic acid, and to discussexplore the relationship between the expression of ATF4 and nonalcoholic fatty liver disease (NAFLD). Mechods The growing cells of L-02 cells line were treated by 20 mg/ml oleic acid which induced the hepatocytes steatosis for 0, 24, 48 and 72 h respectively, and then After co-culture for 24, 48, 72h, the cells were collected and the total RNA and protein were extracted. Reverse transcriptional polymerase chain reaction (RT-PCR) and Western blot analysis were used to detect the expressions of ATF4 gene at different time points. Results With the development of deterioration of steatosis, the expression levels of mRNA were 1.01±0.12, 1.87±0.24, 2.01±0.13, and 1.79±0.19 respectively correspondingly from to 0 hours to 72 hours. The peak of ATF4 mRNA expression was reached at the 48th hours, and then showed the tendency o