青蒿琥酯对Con A诱导小鼠自身免疫性肝损伤的保护作用研究

Protective effects of artesunate against Con A-induced autoimmune liver injury in mice

免疫性肝病是临床上难治性疾病,目前没有特效性药物,因此,研发新的有效干预药物,对防治免疫性肝病有重要的临床价值。为探讨青蒿琥酯（artesunate,Art）对免疫性肝损伤的保护作用,该研究采用不同质量浓度（27,54,108 mg·kg-1）Art灌胃小鼠连续7 d,然后尾静脉注射伴刀豆球蛋白A（concanavalin A,Con A）诱导小鼠肝损伤模型。于造模后8 h采血液和肝组织进行血清转氨酶（ALT,AST）、组织病变、炎性细胞因子以及NF-κB关键蛋白表达水平的检测。结果表明,与模型组相比,Art高浓度组显著降低了模型小鼠的肝指数、ALT和AST水平（P〈0.01）,组织病变也明显减轻,同时降低了促炎细胞因子TNF-α,IFN-γ,IL-6,IL-17的水平而增加了炎性抑制因子IL-10的表达（P〈0.05）,并呈现明显的剂量-效应关系。Western blot法检测结果显示108 mg·kg-1Art可显著抑制NF-κB关键蛋白p-p65和p-IκBα的表达（P〈0.01）。NF-κB特异阻断剂吡咯烷二硫代氨基甲酸酯（PDTC）也可显著抑制p-p65和p-IκBα表达并减轻肝损伤。以上结果说明Art主要通过抑制NF-κB信号通路而对Con A诱导的小鼠免疫性肝损伤发挥保护作用。该研究为青蒿琥酯用于自身免疫性肝损伤的防治提供了参考。

Autoimmune liver disease is a refractory disease clinically,and there is no particularly effective drug at present. Therefore,it is of important clinical value to develop new effective intervention drugs for the prevention and treatment of autoimmune liver disease. In order to investigate the potential protective effect of artesunate（ Art） on concanavalin A（ Con A）-induced autoimmune liver injury,different doses of Art（ 27,54,108 mg·kg-1） were orally administered to mice for consecutive 7 days,respectively. Then the Con A was injected into mice via tail vein to induce liver injury models. 8 h after modeling,the mice were sacrificed. The serum and liver tissue were collected for detecting the level of alanine aminotransferase（ ALT）,and aspartate transaminase（ AST）,liver pathological histopathology,inflammatory cytokines and nuclear factor（ NF-κB） key protein expression level. The results showed that108 mg·kg-1 Art remarkably reduced Con A-induced liver indexes and serum transaminase levels（ ALT and AST） as compared with model group（ P 〈 0. 01）. Meanwhile,the liver histopathological changes were obviously alleviated with a significant decrease of pro-inflammatory cytokine including tumor necrosis factor（ TNF-α）,interferon（ IFN-γ）,interleukin（ IL-6）,IL-17 and a higher increase of anti-inflammatory cytokine IL-10（ P 〈 0. 01）. Western blot results showed that 108 mg·kg-1 Art markedly inhibited the expressions of p-p65 and p-IκBα proteins（ P 〈 0. 01）. The specific inhibitor of NF-κB,pyrrolidinedithiocarbamate（ PDTC） could also significantly inhibit the expressions of p-p65 and p-IκBα with and alleviate liver injuries. Therefore,our results indicated that Art may have a protective action against Con A-induced autoimmune liver injury mainly by suppressing NF-κB signal pathway in mice. The study provides scientific reference for artesunate usage in preventing autoimmune liver injury.