目的 探讨还原型谷胱甘肽片对酒精性脂肪肝（AFLD）斑马鱼模型的保肝功效。方法 选取黑色素等位基因突变型（albino）品系斑马鱼，分别水溶给予还原型谷胱甘肽片500、1 000、2 000、4 000、8 000 μmol/L，同时设置对照组和模型组，除对照组外，利用2%无水乙醇诱导斑马鱼32 h建立AFLD模型，观察记录斑马鱼的死亡情况及表型，确定还原型谷胱甘肽片的最大耐受浓度（MTC）。黑色素等位基因突变型（albino）品系斑马鱼分别给予还原型谷胱甘肽片500、1 000、2 000 μmol/L和阳性对照药RU-21 100 μg/mL，同时设置对照组和模型组，除对照组外建立AFLD模型，采用油红O染色观察斑马鱼肝脏脂肪染色强度（S），计算还原型谷胱甘肽片的保肝功效；H&E染色观察斑马鱼肝脏病理学改变；实时荧光定量PCR（qRT-PCR）检测白细胞介素（IL）-6、高迁移率族蛋白1b（HMGB-1b）和肿瘤坏死因子（TNF）-α mRNA表达。结果 还原型谷胱甘肽片对AFLD模型斑马鱼的MTC为2 000 μmol/L。与对照组比较，模型组S显著增加（P<0.001）；与模型组比较，还原型谷胱甘肽片1 000、2 000 μmol/L组和RU-21组S显著降低（P<0.01、0.001）；还原型谷胱甘肽片500、1 000、2 000 μmol/L和RU-21 100 μg/mL组保肝功效分别为14%、32%、71%和48%。还原型谷胱甘肽片2 000 μmol/L组肝细胞结构正常，脂肪空泡较模型组减少。与对照组比较，模型组IL-6、HMGB-1b和TNF-α mRNA表达显著增加（P<0.05、0.01）；与模型组比较，还原型谷胱甘肽片500、1 000、2 000 μmol/L组IL-6、HMGB-1b和TNF-α mRNA表达显著降低（P<0.01、0.001）。结论 还原型谷胱甘肽片对AFLD具有明显的保肝作用，其机制与下调IL-6、HMGB-1b和TNF-α表达有关。

Objective To investigate the effect and mechanism of glutathione tablets against alcoholic fatty liver disease (AFLD) by constructing a zebrafish model of AFLD. Methods Zebrafish with albino allele mutation were treated with Glutathione Tablets (500, 1 000, 2 000, 4 000 and 8 000 μmol/L) in water solution respectively. Meanwhile, the control group and model group were set up. Except for control group, zebrafish were induced with 2% absolute ethanol for 32 h to establish AFLD model. The mortality and phenotype of zebrafish were observed and recorded. The maximum tolerable concentration (MTC) of Glutathione Tablets was determined. Albino zebrafish were treated with reduced Glutathione Tablets 500, 1 000 and 2 000 μmol/L and positive control drug RU-21 100 μg/mL respectively. The control group and model group were set up at the same time. Except the control group, the AFLD model was established. The liver fat staining intensity (S) of zebrafish was observed by oil red O staining, and the liver protective effect of Glutathione Tablets was calculated; The pathological changes of zebrafish liver were observed by HE staining; Real time fluorescent quantitative PCR (qRT-PCR) was used to detect IL-6, HMGB-1b and TNF-α mRNA expression. Results The MTC of Glutathione Tablets to zebrafish was 2 000 μmol/L. Compared with the control group, S in the model group increased significantly (P < 0.001); Compared with the model group, S of Glutathione Tablets 1 000, 2 000 μmol/L group and RU-21 group were significantly lower (P < 0.01, 0.001); The hepatoprotective effects of Glutathione Tablets 500, 1 000, 2 000 μmol/L and RU-21 100 μg/mL were 14%, 32%, 71% and 48%, respectively. Compared with model group, Glutathione Tablets 2 000 μmol/L group had normal hepatocyte structure and less fat vacuoles. Compared with control group, IL-6, HMGB-1b and TNF-α mRNA expression in model group were significantly higher (P < 0.05, 0.01). Compared with model group, the levels of IL-6, HMGB-1b and TNF-α in Glutathione Tablets 500, 1 000, 2 000 μmol/L group were significantly decreased (P < 0.01, 0.001). Conclusion Glutathione Tablets had obvious protective effect on AFLD, and the mechanism was related to the obvious down-regulation of inflammatory genes such as IL-6, HMGB-1b and TNF-α.

R285.5

南通市科技局项目（JCZ20170）；南通市青年课题A项目（QA2019022）；南通市青年课题（QA2019024）