基于Nrf_(2)通路薏苡附子败酱散联合西药对葡聚糖硫酸钠盐诱导小鼠结肠炎黏膜修复的影响

Protective effect of Yiyi Fuzi Baijiang San combination with Western medicine in treatment of dextran sodium sulfate induces colitis in mice based on Nrf_(2) pathway

目的:研究薏苡附子败酱散联合美沙拉嗪对葡聚糖硫酸钠盐(DSS)诱导的小鼠实验性结肠炎的疗效及肠黏膜损伤修复机制。方法:50只BALB/c雄性小鼠完全随机分为空白组、模型组、西药组、中药组、中西药组,每组10只。DSS诱导慢性实验性结肠炎小鼠模型,西药组给予DSS 7 d后,每天分别给予0.4 g/kg美沙拉嗪灌胃治疗;中药组给予DSS 7 d后,每天分别给予5.24 g/kg薏苡附子败酱散灌胃治疗;中西药组给予DSS 7 d后,每天分别给予5.24 g/kg薏苡附子败酱散和0.4 g/kg美沙拉嗪联合灌胃治疗。30 d后处死全部小鼠,观察记录疾病活动指数(DAI)评分,苏木精-伊红染色法观察组织病理学改变及记录结肠黏膜损伤指数(CMDI)评分,采用ELISA法测定肠组织中髓过氧化物酶(MPO)的活性,采用Western Blot法检测肠组织中红系衍生核因子相关因子2(Nrf_2)和紧密连接蛋白-1(claudin-1)、紧密连接蛋白-2(claudin-2)蛋白表达。结果:与空白组比较,模型组DAI和CMDI评分、MPO活性显著性升高(P<0.05),claudin-2、Nrf_2蛋白表达均显著性上调(P<0.05),claudin-1蛋白表达均显著性下调(P<0.05);与模型组比较,各给药组对以上指标均有不同程度的改善和调控,尤以中西药组最为明显,其明显降低CMDI评分和claudin-2蛋白的过度表达(P<0.05),改善性上调claudin-1蛋白、Nrf_2蛋白(P<0.05)。结论:薏苡仁附子败酱散联合美沙拉嗪对DSS诱导的小鼠慢性实验性结肠炎及结肠病变具有明显的抑制作用,效应明显优于单独薏苡仁附子败酱散组和美沙拉嗪灌胃组。二药联合通过上调Nrf_2蛋白的表达而有效改善抗氧化应激的防御机制。另一方面,其调控病变结肠组织claudin-1/claudin-2蛋白表达,改善慢性溃疡性结肠炎(UC)小鼠结肠黏膜的通透性。这可能是薏苡仁附子败酱散联合美沙拉嗪治疗UC黏膜损伤修复的主要机制。

Objective:To study the effects and mechanism of damaged intestinal mucosal repair of Yiyi Fuzi Baijiang San combination with Mesalazine on the treatment of chronic experimental colitis mice induced by dextran sulphate sodium salt(DSS).Methods:Totally 50 BALB/c male mice were randomly divided into a normal group,a DSS(model)group,a Mesalazine+DSS group(western medicine group),a Yiyi Fuzi Baijiang San+DSS group(Chinese medicines group),a Yiyi Fuzi Baijiang San+Mesalazine+DSS group(Chinese and western medicine group),10 in each group.DSS induced a chronic experimental colitis mouse model.western medicine group were daily treated with 0.4 g/kg Mesalazine after giving DSS for 7 days,Chinese medicines group were daily treated with 5.24 g/kg Yiyi Fuzi Baijiang San after giving DSS for 7 days,Chinese and western medicine group were daily treated with 0.4 g/kg Mesalazine and 5.24 g/kg Yiyi Fuzi Baijiang San after giving DSS for 7 days.All mice were sacrificed after 30 days,The disease activity index(DAI)Score were recorded,The histological features of organization were observed and The colonmucosa damage index(CMDI)were recorded,The activity of MPO in the organization were detected by ELISA,The protein expressions of NF-E2-related Factor2(Nrf2)and claudin-1,claudin-2 in the organization were detected by Western-Blot.Results:Compared with the normal group,The DAI score,CMDI score and MPO activity increased in the DSS model group(all P<0.05),The protein expressions of claudin-2 and Nrf2were significantly up-regulated(all P<0.05),and the Claudin-1 protein expression was significantly down-regulated(P<0.05).Compared with the model group,The above indexes were improved and regulated by each dose group,especially the Chinese and western medicine group were the most obvious,It significantly reduced the CMDI score and the overexpression of claudin-2 protein(all P<0.05),In addition the protein expressions of claudin-1 and Nrf2protein was improved by Up-Regulation(all P<0.05).Conclusion:Yiyi Fuzi Baijiang San combination with western medicine could significantly inhibit inhibited intestinal lesions in a mouse model of chronic experimental colitis induced by DSS,The experimental comprehensive effect was significantly better than that of Yiyi Fuzi Baijiang San group and melalazine group alone.on the one hand,Yiyi Fuzi Baijiang San combination with Mesalazine can upregulate the expression of Nrf2on ulcerative colitis.on the other hand,Yiyi Fuzi Baijiang San combination with Mesalazine can improve the permeability of colonic mucosa in mice with chr.onic ulcerative colitis by regulating the expression of claudin-1/claudin-2 proteins in diseased colon tissues.This may be the main mechanism of Yiyi Fuzi Baijiang San combination with Mesalazine in the treatment of ulcerative colitis mucosal injury repair.