圣草酚对DSS诱导的大鼠溃疡性结肠炎的改善作用及机制探讨

Ameliorating effect and mechanism of eriodictyol on DSS-induced ulcerative colitis in rats

目的:探讨圣草酚对葡聚糖硫酸钠(DSS)诱导的大鼠溃疡性结肠炎(UC)的作用及机制。方法:将50只雄性Wistar大鼠随机分成健康对照组(Control)、模型组(DSS)及3个不同浓度剂量组(DSS+圣草酚10、20、40 mg/kg)。造模结束后,评估大鼠疾病活动指数(DAI)、结肠黏膜损伤指数(CMDI);HE染色观察病理形态,统计结肠组织病理学评分(HS);ELISA检测炎症因子的含量,Western blot检测炎症因子相关蛋白的表达;TUNEL观察结肠组织凋亡情况,Western blot检测凋亡相关蛋白的表达;试剂盒检测氧化应激标志物水平;Western blot检测结肠组织中AKT/NF-κB通路的相关蛋白表达。结果:与模型组相比,除DSS+圣草酚10 mg/kg组差异无统计学意义,DSS+圣草酚20、40 mg/kg组DAI、CMDI、HS评分均明显降低;炎症因子iNOS、TNF-α、IL-1β水平明显降低,抗炎因子IL-10水平明显升高;凋亡率及凋亡相关蛋白表达量明显降低(Bax/Bcl-2、cleaved cas3/cas3);MDA、GSH、LDH含量降低,抗氧化酶SOD活性明显升高;p-AKT/AKT、p-P65/P65的蛋白表达量明显下调。结论:圣草酚能够有效缓解DSS诱导的大鼠UC损伤,其机制可能与下调AKT/NF-κB通路相关蛋白的磷酸化表达,改善炎症水平,减少细胞凋亡,提高大鼠抗氧化能力,缓解氧化应激反应等有关。

Objective:To investigate the effect and mechanism of eriodictyol on dextran sodium sulfate(DSS)-induced ulcerative colitis(UC)in rats.Methods:Fifty male Wistar rats were randomly divided into healthy control group(Control),model group(DSS)and three different concentration dose groups(DSS+Eriodictyol 10,20,40 mg/kg). After model,evaluated the rat disease activity index(DAI)and colonic mucosal injury index(CMDI);HE staining was used to observe the pathological form,statistical colonic histopathological score(HS);ELISA was used to detect the content of inflammatory factors,Western blot was used to detect inflammatory factors the expression of related proteins;TUNEL was used to observe the apoptosis of colon tissue,Western blot was used to detect the expression of apoptosis-related proteins;the kit was used detect the level of oxidative stress markers;Western blot was used to detect the expression of related proteins in the AKT/NF-κB pathway in colon tissue.Results:Compared with the model group,except for the DSS+eriodictyol 10 mg/kg group,there was no significant difference. The DSS+eriodictyol 20,40 mg/kg group DAI,CMDI and HS scores were significantly reduced;levels of inflammatory factors iNOS,TNF-α,IL-1β were significantly reduced,and the level of anti-inflammatory factor IL-10 was significantly increased;the rate of apoptosis and the expression of apoptosis-related proteins were significantly reduced(Bax/Bcl-2,cleaved cas3/cas3);the content of MDA,GSH,LDH were decreased,the antioxidant enzyme SOD activity was significantly increased;the protein expressions of p-AKT/AKT and p-P65/P65 were significantly downregulated.Conclusion:Eriodictyol can effectively alleviate the damage of DSS-induced UC in rats,whose mechanism may be related to down-regulating the phosphorylation expressions of AKT/NF-κB pathway related proteins,improving inflammation,reducing cell apoptosis,and increasing rats antioxidant ability,alleviation of oxidative stress and so on.