脂联素、抵抗素和Apelin在脂肪变肝细胞中的表达及意义

Expression and significance of adiponectin,resistin and Apelin in hepatocyte steatosis

目的:研究脂肪细胞因子脂联素、抵抗素和Apelin在油酸诱导的肝细胞脂肪变性过程中的作用及意义。方法:以常规培养的人正常肝细胞株L02做为对照组,采用油酸诱导L02细胞脂肪变性,建立非酒精性脂肪变性肝细胞模型,设置油酸诱导24、48、72 h各时相点为实验组。用油红O染色镜下观察对照组和实验各组肝细胞形态学变化,用试剂盒检测肝细胞内甘油三酯(triglyceride,TG)水平,并用实时荧光定量PCR与Western blot检测脂联素、抵抗素和Apelin m RNA和蛋白的表达。结果:24 h肝细胞出现脂肪变性,72 h脂肪变性最严重;各实验组肝细胞内TG含量较对照组均升高,差异有统计学意义(LSD-t值和P值分别为8.477、0.000;17.622、0.000;39.251、0.000),72 h增高最明显;实验组脂联素m RNA和蛋白的表达量均降低,都于24 h开始降低,72 h表达最低,与对照组相比差异有统计学意义(LSD-t值和P值分别为8.240、0.000,16.940、0.000,18.292、0.000;13.283、0.000,21.586、0.000,27.030、0.000);实验组抵抗素m RNA的表达上调,于24 h开始增强,72 h表达最强,与对照组相比差异有统计学意义(LSD-t值和P值分别为2.480、0.038;12.003、0.000;24.080、0.000),而其蛋白质的表达上调,于48 h开始增强,72 h表达最高,与对照组比较差异有统计学意义(LSD-t值和P值分别为8.854、0.000;13.301、0.000);实验组Apelin m RNA和蛋白的表达均上调,都于24 h开始增强,72 h表达最高,与对照组相比差异有统计学意义(LSD-t值和P值分别为2.503、0.000,9.720、0.000,13.720、0.000;5.629、0.000,8.251、0.000,15.953、0.000)。结论:油酸诱导的肝细胞脂肪变性模型中脂肪细胞因子脂联素表达下调、抵抗素和Apelin表达上调,提示脂肪细胞因子脂联素、抵抗素和Apelin的表达的调控共同参与了肝细胞非酒精性脂肪变性。

Objective：To explore the expression and significance of adipocytokines adiponectin,resistin and Apelin in hepatocyte steatosis induced by oleic acid. Methods：Routinely cultured human L02 cells were used as controls;L02 cells were induced by oleic acid for 24,48,72 h respectively to establish nonalcoholic hepatocyte steatosis models（model groups）. Lipid droplets in cells were observed by oil red staining and the levels of triglyceride were measured with analyzed kit. The expressions of m RNA and protein of adiponectin,resietin and Apelin were measured by q PCR and Western blot. Results：Oil red O staining showed that hepatocyte steatosis occurred at 24 h after L02 cells being treated with oleic acid and hepatocyte steatosis increased greatly at 72 h. Triglyceride in hepatocytes of model groups also increased compared with that of control group. In model groups,the expression levels of adiponctin m RNA and protein decreased at 24 h and declined remarkably at 72 h,compared with those of control groups. The expression levels of resistin m RNA increased at 24 h and elevated remarkably at 72 h in model groups,compared with those in control group. The expression levels of resistin protein increased at 48 h and elevated remarkably at 72 h in model groups,compared with those in control group. The expression levels of Apelin m RNA and protein increased at 24 h and elevated remarkably at 72 h in model groups,compared with those in control group. Conclusion：The expressions of adiponectin are down-regulated while those of resistin and Apelin are up-regulated in hepatocyte steatosis model,indicating the regulation of adipocytokines. Adiponectin resistin and Apelin may be jointly involved in hepatocyte steatosis.