土家药竹节参多糖通过Nrf2-ARE信号通路对四氯化碳致大鼠急性肝损伤的抗氧化保护作用

Anti-oxidative protection of Panax japonicus polysaccharide on CCl4-induced acute liver injury in rats based on Nrf2-ARE signaling pathway

目的基于核因子E2相关因子2(nuclear factor E2 related factor 2,Nrf2)-抗氧化反应元件(antioxidant response element,ARE)信号通路研究竹节参多糖(Panax japonicus polysaccharide)保护四氯化碳(carbon tetrachloride,CCl_(4))所致大鼠急性肝损伤的作用机制。方法通过CCl_(4)间隔诱导法建立急性肝损伤大鼠模型,给予水提醇沉法制备的竹节参多糖治疗7 d后取材,通过苏木素-伊红(HE)染色观察肝组织病理变化;全自动生化仪检测大鼠血清中肝功能相关指标;ELISA法检测肝组织氧化应激水平;免疫组化法观察大鼠肝组织Nrf2表达;Western blotting测定大鼠肝组织Nrf2-ARE通路相关蛋白表达。结果与模型组比较,竹节参多糖组大鼠血清中肝功能相关指标呈不同程度的降低(P<0.05、0.01),肝组织中Nrf2-ARE信号通路明显激活(P<0.01),肝脏中脂类过氧化产物丙二醛(malondialdehyde,MDA)水平降低(P<0.01),抗氧化酶活性明显升高(P<0.05、0.01),细胞质中氧化应激效应物Nrf2阳性的细胞数有所减少(P<0.01),大鼠肝脏损伤程度明显减轻。结论竹节参多糖可激活Nrf2-ARE信号通路,增强机体的抗氧化酶系表达,减轻CCl_(4)引起的氧化损伤。

Objective To study the protective mechanism of Panax japonicus polysaccharide(PSPJ)against carbon tetrachloride(CCl_(4))induced acute liver injury in rats through nuclear factor E2 related factor 2(Nrf2)-antioxidant response element(ARE)signal pathway.Methods A rat model of acute liver injury was established using the CCl_(4) interval induction method.After 7 d of treatment with PSPJ prepared by water extraction and alcohol precipitation method,samples were taken and liver tissue pathological changes were observed by hematoxylin eosin(HE)staining;Liver function related indicators in serum of rat were detected by fully automated biochemical analyzer;ELISA method was used to detect oxidative stress level in liver tissue;Immunohistochemical method was used to observe the expression of Nrf2 in liver tissue of rats;Western blotting was used to determine the expressions of Nrf2-ARE pathway related proteins in liver tissue of rats.Results Compared with model group,liver function related indicators in serum of rats treated with PSPJ showed varying degrees of decrease(P<0.05,0.01),Nrf2-ARE signaling pathway was significantly activated in liver tissue(P<0.01),level of lipid peroxidation product malondialdehyde(MDA)in liver was reduced(P<0.01),and activity of antioxidant enzymes was significantly increased(P<0.05,0.01),the number of cells with Nrf2 positive oxidative stress effector in cytoplasm was decreased(P<0.01),and the degree of liver damage in rats was significantly reduced.Conclusion PSPJ can activate the Nrf2-ARE signaling pathway,enhance the expression of antioxidant enzymes in body,and alleviate oxidative damage caused by CCl_(4).