基于沉默信息调节因子1调控内质网应激探讨化浊解毒消痈方对溃疡性结肠炎小鼠的保护作用

Protective effect of Huazhuo Jiedu Xiaoyong Formula in mice with ulcerative colitis based on the regulation of endoplasmic reticulum stress by SIRT1

目的:基于沉默信息调节因子1(SIRT1)调控内质网应激(ERS)探讨化浊解毒消痈方对溃疡性结肠炎(UC)小鼠的保护作用。方法:将C57BL/6小鼠随机分为正常组,模型组,美沙拉嗪组,化浊解毒消痈方高、中、低剂量组,每组10只。葡聚糖硫酸钠(DSS)建立UC大鼠模型,造模成功后分别予化浊解毒消痈方药28.68、14.34、7.17 g/kg灌胃,美沙拉嗪0.45 g/kg灌胃,模型组及正常组以等体积0.9%氯化钠溶液灌胃,均连续7 d。观察各组小鼠的一般情况,观察各组大鼠疾病活动指数(DAI)及结肠组织SIRT1、葡萄糖调节蛋白78(GRP78)、C/EBP同源蛋白(CHOP)、半胱氨酸蛋白酶-3(Caspase-3)蛋白和mRNA表达水平。结果:与模型组比较,化浊解毒消痈方各剂量组可显著降低UC大鼠DAI评分(P<0.05)。与正常组比较,模型组小鼠结肠组织SIRT1 mRNA和蛋白表达显著降低(P<0.05);经治疗后,各药物治疗组SIRT1 mRNA和蛋白表达升高(P<0.05),其中化浊解毒消痈方高剂量组效果较佳,与美沙拉嗪组效果相当。与正常组比较,模型组结肠组织中ERS标志性分子GRP78和凋亡相关分子CHOP、Caspase-3 mRNA和蛋白表达均显著升高(P<0.05);经治疗后,各药物治疗组表达均显著下调(P<0.05),其中化浊解毒消痈方高剂量组效果较佳,与美沙拉嗪组效果相当。结论:化浊解毒消痈方治疗UC的作用机制可能是通过上调SIRT1的表达,进而抑制内质网过度应激,抑制肠上皮细胞凋亡,减轻肠道黏膜损伤,从而发挥治疗UC的作用。

Objective:To explore the protective effect of Huazhuo Jiedu Xiaoyong Formula in mice with ulcerative colitis(UC)based on the regulation of endoplasmic reticulum stress(ERS)by silent information regulator 1(SIRT1).Methods:C57BL/6 mice were randomly divided into normal group,model group,Mesalazine group,and Huazhuo Jiedu Xiaoyong Formula high dose,medium dose and low dose group,with 10 mice in each group.The animal model of the UC was established with dextran sulfate sodium(DSS).After successful molding,Huazhuo Jiedu Xiaoyong Formula 28.68,14.34,7.17 g/kg was administered,and Mesalazine 0.45 g/kg was fed.The model group and normal group were fed with 10 mL/kg of 0.9%sodium chloride solution every day for 7 d continuously.Changes of disease activity index(DAI)score was observed,and the protein and mRNA expression levels of SIRT1,GPR78,CHOP,Caspase-3 in colon tissue of mice in each group were observed.Results:Compared with the model group,Huazhuo Jiedu Xiaoyong Formula three dose groups cowd can significantly reduce the DAI score(P<0.05).Compared with the normal group,the mRNA and protein expression of SIRT1 in the colon tissue of the mice in the model group significantly decreased(P<0.05).After treatment,the mRNA and protein expression of SIRT1 increased in each drug treatment group(P<0.05).Among them,the high dose Huazhuo Jiedu Xiaoyong Formula group had the better effect,which was equivalent to the Mesalazine group.Compared with the normal group,the expression levels of GRP78,a marker of ERS,apoptosis-related molecules CHOP,and Caspase-3 in the colon tissue of the model group significantly increased(P<0.05).After treatment,the expression of each drug treatment group significantly decreased(P<0.05),and the high dose Huazhuo Jiedu Xiaoyong Formula group had the better effect,which was equivalent to the Mesalazine group.Conclusion:The mechanism of Huazhuo Jiedu Xiaoyong Formula in treating UC may be through up regulating the expression of SIRT1,thus inhibiting the excessive stress of endoplasmic reticulum,inhibiting the apoptosis of intestinal epithelial cells,and alleviating the damage of intestinal mucosa,so as to play the role of treating UC.