乙酰半胱氨酸抗非酒精性脂肪性肝炎的体外活性研究

Study of N-acetylcysteine's Anti-Nonalcoholic Steatohepatitis Activity in Vitro

关键词:  乙酰半胱氨酸  非酒精性脂肪性肝炎  胰岛素抵抗  炎症反应

Key words:  N-acetylcysteine  nonalcoholic steatohepatitis  insulin resistance  inflammatory response

摘要:

目的 研究乙酰半胱氨酸（N-acetylcysteine，NAC）对游离脂肪酸（free fatty acid，FFA）诱导的L02细胞的非酒精性脂肪性肝炎（nonalcoholic steatohepatitis，NASH）模型的炎症反应和胰岛素抵抗的活性。方法 用FFA处理L02细胞，建立NASH细胞模型；MTT法检测NAC对细胞生长抑制率的影响，并依此优选3个安全给药浓度；油红染色法观察细胞内的脂质沉积情况；Elisa法检测细胞的IL-8、TNF-α和葡萄糖转运蛋白-4（glucose transporter，GLUT-4）含量；Western blot及Elisa法检测细胞的胰岛素受体（insulin receptor，IR）含量。结果 NAC在10600.0μg·mL^-1的最大给药浓度时其细胞生长抑制率仅为34.10%，因此设定100，150和250μg·mL^-13个给药浓度；NAC各给药组较模型组82.45%的细胞脂变率，分别下降至61.77%，68.08%和79.28%，差异具有统计学意义（P<0.01）且呈浓度依赖性；NAC各给药组的GLUT-4和IR含量较模型组均显著提高（P<0.01）；NAC各给药组的TNF-α和IL-8含量较模型组均显著下降（P<0.01）。结论 NAC的细胞毒性极小，可不同程度地降低NASH细胞的脂变率，尤其可改善模型细胞的胰岛素抵抗及炎症反应水平，表现出在治疗NASH方面的潜在价值。

Abstract:

OBJECTIVE To investigate the effects of N-acetylcysteine(NAC) on the inflammatory response and insulin resistance in the model of nonalcoholic steatohepatitis(NASH) induced by free fatty acid(FFA) in L02 cells.METHODS L02 cells were treated with FFA to establish NASH cell model; The growth inhibition ratio of NAC was measured by MTT to select the safe drug concentration. The fat deposition in cells was observed by oil-red staining method. The content of IL-8, TNF-α and glucose transporter-4(GLUT-4) were tested by Elisa; And the content of cells' insulin receptor(IR) were tested by Western blot and Elisa.RESULTS The growth inhibition ratio was only 34.10% under the NAC's maximum concentration of 10 600.0 μg·mL^-1, so 3 treatment groups were seted with the NAC concentration of 100, 150 and 250 μg·mL^-1. The steatosis ratio of the model group was up to 82.45%, but the treatment groups' ratio felled to 61.77%, 68.08% and 79.28% significantly(P<0.01) with a concentration dependence; The treatment groups' concentration of GLUT-4 and IR was increased significantly(P<0.01) compared with the model group, but significant concentration decreases of IL-8 and TNF-α were observed(P<0.01).CONCLUSION NAC has a tinny cytotoxicity. It can decrease NASH cells' steatosis ratio and especially improve their insulin resistant and inflammatory response level, showing potential value on NASH treatment.