瘦素对脂变肝细胞脂蛋白脂酶基因表达的影响(1)
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【摘要】 目的 研究瘦素对脂变肝细胞甘油三酯含量、脂滴形成和脂蛋白脂酶基因表达的影响。方法 以离体人肝L-02细胞株为研究对象。用含10%医用脂肪乳注射液和10%胎牛血清的RPMI-1640的完全培养基孵育人肝L-02细胞24小时,制备肝细胞脂肪变性模型。实验分为正常肝细胞组(A组)、脂变肝细胞模型组(B组)、瘦素组(10-7mol/L)、阳性对照组(C组)(即脂变肝细胞加入吉非罗齐,使其终浓度为1000mg/L);孵育24小时后,油红O染色观察细胞形态和细胞内脂滴的形成情况,高效液相色谱法检测细胞内甘油三酯的含量,半定量RT-PCR法检测LPL mRNA 水平的表达。实验数据以x±s表示,用SPSS11.0统计软件对数据进行分析,以P<0.05作为差异显著性标准。结果 用脂肪乳注射液孵育人肝L-02细胞24小时后,经油红O染色后倒置显微镜观察,人肝L-02细胞浆内见大量红色的小泡性脂滴;高效液相色谱法检测示细胞内甘油三酯含量明显增高,成功的制备肝细胞脂肪变性模型。加入瘦素(10-7mol/L,)孵育24小时后,油红O染色示胞浆内脂滴与模型组比较明显减低;高效液相色谱法结果发现瘦素能降低细胞内甘油三酯含量;半定量RT-PCR法检测各组细胞的LPL mRNA水平,结果显示:瘦素处理组与模型组相比,LPLmRNA表达明显上升,有统计学意义(P<0.05)。结论 1. 予以干预脂变肝细胞,瘦素能降低人L-02脂变肝细胞内甘油三酯的含量;2. 瘦素能促进、LPLmRNA的表达增强。
【关键词】 瘦素;人肝L-02细胞;甘油三酯;脂蛋白脂酶
【中图分类号】R329.2+8 【文献标识码】 B 【文章编号】1005-0515(2010)007-022-04
Effects of Leptin on the expression of lipoprotein lipase in human L-02 hepatocyte
ABSTRACT
Objects: To observe the effects of leptin on triglyceride levels , the formation of lipid droplets and the expresion of lipoprotein lipase(LPL)mRNA in human L-02 steatosis hepatocyte. Methods: To establish a model of hepatocyte steatosis, we incubate the normal human L-02 hepatocyte in RPMI-1640 complete medium with 10% fetal bovine serum plus 10% medical fat emulsion injection for 24 hours. The experiment is divided into 7 groups: The first group is normal hepatocytes(A group); the second group is fatty change hepatocytes(B group); the third group(C group) is added leptin to the fatty changed hepatocytes to make the concentration to be 10-7mol/L. Group 4 is the positive control group(D group), i.e Gemifibrozil is added to the hepatocyte with fatty change to make the concentration to be 1000 mg/L. Incubate these human L-02 hepatocyte for another 24 hours. And observe the formation of cellular morphology and lipid droplets in the cells under oil red O stain and determin the intracellular triglyceride levels through high performance liquid chromatography (HPLC) ......
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