基于Nrf2/HO-1信号通路的红萸饮抑制氧化应激保护葡聚糖硫酸钠诱导的炎症性肠病小鼠的调控作用探讨

Effect of Hongyuyin inhibiting oxidative stress on inflammatory bowel disease induced by dextran sodium sulfate in mice based on Nrf2/HO-1 signaling pathway

目的 评价核因子-E2相关因子(NRF2)/血红素氧合酶-1(HO-1)信号通路在红萸饮抑制氧化应激保护葡聚糖硫酸钠诱导的炎症性肠病小鼠中的作用。方法 将50只C57BL/6小鼠随机分为正常组、模型组、红萸饮低(8.95g/kg)、中(17.89g/kg)、高(35.76g/kg)剂量组,每组10只。除正常组外,其他组以2.5%葡聚糖硫酸钠自由饮用建立炎症性肠病模型。红萸饮组分别予低、中、高剂量红萸灌胃治疗,正常组、模型组予等体积生理盐水灌胃,连续7d。每日记录小鼠体质量、粪便、精神状态等基本情况,统计疾病活动指数行疾病活动指数(DAI)评分。第14天处死所有小鼠,观察小鼠结直肠长度及组织病理学变化;生化试剂盒检测小鼠肠道组织中CAT、GSH-Px的活性水平;Western Blot法、RT-qPCR法检测小鼠肠道组织中NRF2、HO-1蛋白及mRNA表达。结果 与正常组相比,模型组小鼠DAI评分升高,肠壁组织破坏,CAT、GSH-Px活性和NRF-2、HO-1表达下降(P<0.01);与模型组相比,红萸饮各给药组小鼠结肠组织破坏得到改善,CAT、GSH-Px活性上升,其中红萸饮中剂量组小鼠结肠组织中Nrf-2、HO-1蛋白和基因表达上升最为明显(P均<0.01)。结论 红萸饮可通过抑制氧化应激调控NRF2/HO-1信号通路治疗DSS导致的小鼠IBD。

Objective To explore the protective effect of Hongyuyin inhibiting oxidative stress on inflammatory bowel disease in⁃duced by dextran sodium sulfate in mice based on Nrf2/HO-1 signaling pathway.Methods Fifty mice were randomly divided in⁃to normal group,model group and low,medium and high dosage groups of Hongyuyin(8.95,17.89 and 35.76g/kg),with 10 mice in each group.Establish models for other groups except for the normal group.Inflammatory bowel disease(IBD)model was established by drinking 2.5%DSS freely.Hongyuyin group was given low,medium and high doses of Hongyuyin by gavage re⁃spectively,and normal group and model group were given equal volume of normal saline by gavage for 7 days.The body weight,feces and mental state of mice were recorded every day,and the disease activity index(DAI)was scored.On the 14th day,all mice were killed,the length of colorectal cancer was recorded,and histopathological changes were observed by HE staining.The level of CAT and GSH-Px in colon tissue was detected by biochemical kit.The protein and mRNA expressions of Nrf2、HO-1 in colon tissue were detected by Western blot and Quantitative Real-time PCR.Results Compared with the normal group,DAI score,colonic histopathology score,of the model group were increased(P<0.01),the level of CAT and GSH-Px and the ex⁃pression of Nrf2、HO-1 protein and mRNA were all decreased(P<0.01),the colon length was shortened(P<0.01),and the intestinal wall tissue was damaged.Compared with the model group,the DAI score,colonic histopathology score were all de⁃creased in each dosage group of Hongyuyin,the length of colon and the level of CAT and GSH-Px and the expressions of Nrf2、HO-1 protein and mRNA were all increased(P<0.01),and the damage of intestinal wall tissue was improved.Conclusion Hongyuyin can regulate Nrf2/HO-1 signaling pathway by inhibiting oxidative stress to treat IBD induced by DSS in mice.