中文摘要：

目的研究胰岛素样生长因子-1（IGF-1）及胰岛素样生长因子结合蛋白-3（IGFBP-3）基因在非酒精性脂肪变性肝细胞模型中的表达变化及其意义。方法用油酸诱导永生化人肝细胞（IHH）建立非酒精性脂肪性肝病（NAFLD）细胞模型，油红O染色和细胞内甘油三酯含量检测观察细胞脂肪变情况。IHH细胞分为对照组和NAFLD组，对照组细胞以DMEM／F12培养基培养，NAFLD组给予油酸0．5mmol／L处理72h。采用逆转录酶-聚合酶链反应及Westemblot和免疫荧光染色方法检测IGF-1和IGFBP-3在两组细胞中的mRNA及蛋白质表达变化。组间均数比较采用t检验。结果0．5mmol／L油酸可成功诱导IHH细胞脂肪变性，油红O染色显示细胞内脂肪滴明显增多，细胞内甘油三酯含量从对照组的（150．2±15．6）μg／mg升高到（275．7±27．2）ug／rag（t=21．67，P〈0．01）。油酸诱导后，NAFLD组细胞中IGF-1和IGFBP-3的mRNA相对表达量（分别为0．76±0．04和1．58±0．93）均较对照组（分别为4．82±1．51和5．41±1．37）明显下降，t值分别为17．915和12．893，P值均〈0．01；IGF-1和IGFBP-3的蛋白质相对表达量（分别为1．00±0．29和0．65±0．36）也较对照组（分别为2．56±0．71和1．23±0．91）明显下降，t值分别为29．17和32．12，P值均〈0．01。免疫荧光染色结果也证实NAFLD组细胞中IGF-1和IGFBP-3的蛋白质表达较对照组明显下降。结论非酒精性脂肪变性肝细胞模型的IGF-1和IGFBP-3表达下降，为深入研究临床上部分非酒精性脂肪肝病儿童身高受限的机制提供了实验基础。

英文摘要：

Objective To investigate the expression of IGF-1 and IGFBP-3 in nonalcoholic fatty steatosis hepatocyte models induced by oleic acid. Method Nonalcoholic fatty steatosis hepatocyte models induced by oleic acid on immortalized human hepatocyte, Oil red O staining and intracellular triglycerides were detected for observing the situation of IHH ceils fatty degeneration. IHH cells were divided into control group, NAFLD group, which the control group cultured in DMEM/F12 medium, NAFLD group were treated with oleic acid, 0.5 mmol / L treatment for 72 h. The expression of mRNA and protein of IGF-1 and IGFBP3 were measured by immunofluorescent staining, Western blot and RT-PCR methods. Between the two groups were compared using the t- test. Results The steatosis models of the hepatocytes were established successfully with 0.5 mmol/L oleic acid. Lipid droplets were observed through Oil red O staining. The level of hepatocyte TG was increased （275.7 ± 27.2） μg/mg from （150.2 ± 15.6） μg/mg （t = 21.67, P 〈 0.01）. Compared with the control group, the mRNA of IGF-1 （0.76 ± 0.04 vs 4.82 ± 1.51, t = 17.915, P 〈 0.01）, IGFBP-3 （1.58 ± 0.93 vs 5.41 ± 1.37, t= 12.893,P〈 0.01） and protein expression of IGF-1 （1.00 ± 0.29 vs 2.56 ± 0.71, t= 29.17, P 〈 0.01）, IGFBP-3 （0.65 ± 0.36 vs 1.23 ± 0.91, t = 32. 12, P 〈 0.01） significantly decreased in oleic acidtreated group. The results of immunofluorescence staining also confirm the significantly decreased protein expression oflGF-1 and IGFBP-3 in NAFLD group compared with control group. Conclusion The expression of IGF-1 and IGFBP-3 decreased in nonalcoholic fatty steatosis hepatocyte models, which will provide the experimental basis for the further study of the mechanism of the limited height of some children with non- alcoholic fatty liver disease in clinical.