复方苦参汤对溃疡性结肠炎小鼠结肠组织中炎症因子及自噬水平的影响  

Effect of Compound Sophorae Decoction on inflammatory factors and autophagy levels in colonic tissue of mice with ulcerative colitis

作　　者：邓双娇 左冬梅 陈倩云 刘与进 李华蓉 DENG Shuangjiao;ZUO Dongmei;CHEN Qianyun;LIU Yujin;LI Huarong(Department of Integrated Traditional Chinese and Western Medicine,Union Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430022,China;Department of Traditional Chinese Medicine,Jingshan Union Hospital,Huazhong University of Science and Technology,Jingmen,Hubei 431800,China)

机构地区：[1]华中科技大学同济医学院附属协和医院中西医结合科,湖北武汉430022 [2]华中科技大学协和京山医院中医科,湖北荆门431800

出　　处：《上海中医药杂志》2023年第9期63-69,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82205058,81904018);华中科技大学同济医学院附属协和医院2021年自由创新预研基金(F0160200421003116)。

摘　　要：目的观察复方苦参汤对溃疡性结肠炎(UC)小鼠结肠组织中炎症因子及自噬水平的影响,并探讨其作用机制。方法将20只SPF级C57BL/6雄性小鼠随机分为正常组、模型组、复方苦参汤组、美沙拉嗪组,每组5只。适应性喂养7 d后,除正常组外,其余3组予以3%葡聚糖硫酸钠(DSS)自由饮用7 d制备UC小鼠模型。同时在造模第1天开始,正常组和模型组每日均以质量分数为0.9%的氯化钠溶液灌胃,复方苦参汤组与美沙拉嗪组分别予复方苦参汤(7.28 g/kg)、美沙拉嗪(0.52 g/kg)灌胃,每日1次,连续灌胃7 d。每天记录小鼠体质量、大便性状及便血情况,进行疾病活动指数(DAI)评分。末次灌胃结束后,取各组小鼠结肠组织,测量结肠长度,肉眼观察肠黏膜炎症损伤程度进行结肠黏膜损伤指数(CMDI)评分。采用苏木精-伊红染色(HE)法观察结肠组织病理变化,酶联免疫吸附试验(ELISA)检测白介素-1β(IL-1β)、白介素-18(IL-18)含量;免疫印迹(Western blot)法及实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测自噬相关分子蛋白及mRNA表达。结果与正常组比较,模型组大鼠DAI评分、CMDI评分、促炎因子IL-1β和IL-18,以及泛素结合蛋白P62(P62)分子表达升高(P<0.05),结肠长度缩短(P<0.05),而抑炎因子白介素-4(IL-4)、白介素-10(IL-10)的表达下降(P<0.05),Bcl-2同源结构域(Beclin1)蛋白、微管相关蛋白1A/1B轻链3(LC3)蛋白和其mRNA的表达水平明显降低(P<0.05)。与模型组比较,复方苦参汤组与美沙拉嗪组的DAI评分、CMDI评分、促炎因子IL-1β和IL-18,以及P62分子均表达降低(P<0.05),结肠长度显著增加(P<0.05),而抑炎因子IL-4和IL-10,以及Beclin1、LC3的表达水平明显升高(P<0.05)。与美沙拉嗪组比较,复方苦参汤组的CMDI评分增加(P<0.05),IL-1β、IL-18及P62的表达均升高(P<0.05),而IL-4、IL-10,Beclin1、LC3的表达降低(P<0.05),同时结肠长度以及DAI评分差异无统计学意义(P>0.05)。�Objective To observe the effect of Compound Sophorae Decoction(CSD)on inflammatory factors and autophagy in colon tissue of mice with ulcerative colitis(UC),and to explore its mechanism of action.Methods Twenty SPF C57BL/6 male mice were randomly divided into normal group,model group,CSD group and mesalazine group,with 5 mice in each group.After one week of adaptive feeding,UC mouse models were prepared by giving 3%dextran sulfate sodium(DSS)ad libitum for 1 week to mice in all three groups except the normal group.From the first day of modeling,the normal and model groups were intragastrically administered with 0.9%sodium chloride solution by mass fraction daily,and the CSD group and the mesalazine group were intragastrically administered once daily for 1 week with CSD(7.28 g/kg)and mesalazine(0.52 g/kg)respectively.Body weight,stool characteristics and hematochezia were recorded every day,and disease activity index(DAI)scores were measured.After the final gavage,the colonic tissues of mice in each group were taken to measure the length of the colon,and the degree of intestinal mucosal inflammatory injury was macroscopically observed for CMDI scoring.HE staining was used to observe the histopathological changes,ELISA was used to detect the contents of IL-1βand IL-18,and Western blot and RT-qPCR were used to detect autophagy-related molecule protein and mRNA expression.Results According to the inter-group comparison between normal group and model group,the DAI score,CMDI score,the expression of pro-inflammatory cytokines IL-1β,IL-18 and P62 molecules increased(P<0.05),the colon length was shortened(P<0.05),the expression of anti-inflammatory cytokines IL-4 and IL-10 decreased(P<0.05),and the expression levels of Beclin1 and LC3 protein and mRNA decreased significantly(P<0.05)in the model group.According to the inter-group comparison between the model group and the CSD and mesalazine groups,the DAI score,CMDI score,pro-inflammatory cytokines IL-1β,IL-18 and P62 molecules decreased(P<0.05),the colon length signif

关 键 词：溃疡性结肠炎 复方苦参汤 自噬 炎症因子 小鼠模型 中药研究 

分 类 号：R285.5[医药卫生—中药学]