摘要
目的探讨抵抗素在非酒精性脂肪性肝炎（NASH）发病中对肝细胞的致炎作用及其分子机制。方法制作NASH大鼠模型，分别应用荧光实时定量PCR和免疫组织化学染色的方法检测NASH大鼠肝组织抵抗素的表达。体外培养AML-12小鼠肝细胞株，用重组抵抗素或细菌脂多糖处理后48h，检测细胞培养上清液中肿瘤坏死因子仅、白细胞介素6含量的变化，用免疫荧光染色观察肝细胞核因子κB的核转位。采用Student-Newman-Keuls法进行组间比较。结果NASH大鼠肝组织表现出明显脂肪变性、小叶内炎症和窦周纤维化；随造模时间的延长，抵抗素mRNA和蛋白表达逐渐增强，在造模12周及16周，抵抗素mRNA的表达分别为对照组的2．5倍和4．0倍，抵抗素蛋白的表达主要集中在肝组织中央静脉周围。用重组抵抗素处理AML-12细胞48h后，细胞培养上清液中肿瘤坏死因子α、白细胞介素6的浓度分别为（1．856±0．049）pg／ml和（9．463±1．216）pg／ml，与脂多糖处理组[（1．791±0．046）pg／ml和（8．738±1．101）pg／ml]比较，差异无统计学意义，但二者均显著高于正常对照组[（1．310±0．038）pg／ml和（3．260±0．213）pg／ml]，P〈0．01；用重组抵抗素或脂多糖处理AML-12细胞3h观察到细胞核因子κBp65的核转位。结论抵抗素是NASH发病中重要的致炎因子之一，其可以通过核因子κB途径诱导肝细胞产生肿瘤坏死因子α、白细胞介素6等炎性细胞因子，导致肝组织的炎症反应。
Objective To investigate the role and molecular mechanism of resistin in inflammation of hepatocytes in nonalcoholic steatohepatitis. Methods Rat models of NASH were established successfully. The expression of resistin mRNA and protein were examined by quantitative RT-PCR and immunohistolostaining, respectively. The murine hepatocytes AML- 12 were incubated with recombinant resistin or LPS for 48 hours, and the concentration of TNF α, IL-6 in supematant of AML-12 cells were quantified by enzyme linked immunosorbent assay （ELISA）, the nuclear translocation NF- κ B were observed by immunofluorescence. Results The steatosis of hepatocytes, inflammation in the lobule and perisinusoidal fibrosis in livers were found, and the expression of resistin mRNA and protein were increased in livers of rat model of NASH. The expression of resistin mRNA was 2.5 and 4 time higher in 12 weeks and 16 weeks of rat models respectively than that in normal control. The positive staining of resistin protein can be found mainly around the central veins. The concentration ofTNF ct and IL-6 were （1.856 ± 0.049） pg/ml and （9.463± 1.216） pg/ml in supernatant of AML-12 cells 48 hours after recombinant resistin treatment, and （1.791± 0.046） pg/ml, （8.738 ± 1.101） pg/ml 48 hours after LPS treatment. There was no significant difference between them, but both were higher than that in normal control （P 〈 0.01）. The NF- κB p65 nuclear translocation had been observed in AML-12 cells 3 hours after resistin or LPS treatment. Conclusions Resistin can induce the production of TNF α, IL-6 and other inflammatory factors by hepatocytes, and therefore is an important inflammatory factor in NASH.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2009年第9期683-687,共5页
Chinese Journal of Hepatology
基金
河北省自然科学基金（C200800941）
