抵抗素对脂肪变性肝细胞核因子κB和肿瘤坏死因子α表达的影响  

The effect of resistin on nuclear factor-κB and tumor necrosis factor-α expression in hepatic steatosis

作　　者：祁明美[1] 管小琴[1] 朱良荣[1] 王立娟[1] 刘琳[1] 杨云鹏[1] 

机构地区：[1]400016,重庆医科大学病理学教研室 分子医学与肿瘤研究中心

出　　处：《中华肝脏病杂志》2012年第1期40-44,共5页Chinese Journal of Hepatology

基　　金：重庆市教委资助项目

摘　　要：目的探讨抵抗素（resistin）对脂肪变陛肝细胞脂质代谢及核因子κB（NF—κB）、肿瘤坏死因子（TNF）α表达的影响。方法选用软脂酸诱导L02细胞脂肪变性模拟非酒精性脂肪性肝病（NAFLD）体外模型。分C组（正常对照组），P组（模型组：软脂酸20μg／ml）及分别加入重组人抵抗素50μg／L的CR组和PR组。各组给予相应处理72h后，观察细胞的脂肪变并测定细胞培养上清液中甘油三酯（TG）、ALT、AST及γ谷氨酰转移酶（GGT）的浓度；采用半定量RT-PCR及Westem blot法检测细胞胰岛素受体底物2（IRS-2）、NF—κB、TNF-α的基因及蛋白表达水平。根据资料不同应用方差分析或t检验进行统计学分析，P〈0．05为差异具有统计学意义。结果P组、CR组及PR组细胞培养上清液中TG、ALT、AST、GGT的浓度明显高于C组。分别与C组比较，P组、CR组及PR组NF—κBmRNA相对表达量0．54±0．04、0．604-0．04及1．00±0．06，与C组的0．10±0．01比较，f值分别为17．64，22．03，26．06，TNF-αmRNA相对表达量0．58±0．04、0．61±0．06及1．05±0．09，与C组的0．33±0．06比较，t值分别为5．67，5．38，11．64，相对表达量明显升高，而IRS-2mRNA（t值分别为8．19，9．23，20．93）明显降低，且P值均〈0．05，差异有统计学意义。P组与CR组间比较，t值分别为1．75，0．58，2．14，P值均〉0．05，差异无统计学意义。蛋白质表达变化趋势与mRNA变化趋势相一致。结论抵抗素可能通过NF-κB依赖途径对肝细胞脂肪变性胰岛素抵抗发挥促炎作用，从而促进NAFLD的发生与发展。Objective To investigate the potential regulatory role played by the hormone resistin in lipid metabolism and expression of nuclear factor （NF）-κB and tumor necrosis factor （TNF）-α during hepatic steatosis. Methods A non-alcoholic fatty liver disease （NAFLD） cell model was established by treating the normal human hepatic cell line, L02, with palmitic acid. Four research groups of L02 cells were generated： C group （control, no palmitic acid treatment）, P group （NAFLD model, treated with 20 μg/ml palmitic acid）, CR group （C group treated with 50 μg/L recombinant human resistin）, and PR group （P group treated with 50μg/ L recombinant human resistin）. All treatments were carried out for 72 hours. Oil red O staining was used to detect the intracellular changes in lipid drops. Biochemical assays were used to measure triglyeerides （TGs）, alanine aminotrausferase （ALT）, aspartate aminotransferase （AST）, and gamma-glutarnyl transpeptidase （GGT） levels in culture medium. The mRNA and protein expression levels of insulin receptor substrate （IRS）-2, NF- κB, and TNF-α were determined by reverse transcription-polyrnerase chain reaction and Western blot analysis, respectively. Results The TG, ALT, AST, and GGT levels were higher in the P, CR, and PR groups thanin the C group. The NF-κB mRNA level was also higher in the P, CR, and PR groups （Student＇s t = 17.64, 22.03, 26.06 respectively） than in the C group, as was the TNFα mRNA level （ t = 5.67, 5.38, 11.64）, but the IRS-2 mRNA level was lower （ t = 8.19, 9.23, 20.93） （all, P 〈 0.05）. In addition, no significant difference in these mRNA levels were found between the P group and the CR group （NF-κB： t = 1.75, TNFα： t = 0.58, IRS-2： t = 2.14; all, P 〉 0.05）. The detected protein levels of NF-κB, TNFα, and IRS-2 were consistent with the mRNA levels. Conclusion Resistin can promote steatosis in LO2 cells through the NF-κB signaling pathway, thereby contributing to the NAFLD pathogenic p

关 键 词：脂肪肝 非酒精性 抵抗素 NF-ΚB 肿瘤坏死因子Α 

分 类 号：R575.5[医药卫生—消化系统]