基于Nrf2通路探讨溃结宁膏穴位敷贴治疗溃疡性结肠炎的作用机制  

Mechanism of Acupoint Application of Kuijiening Plaster in Treatment of Ulcerative Colitis Based on NRF2 Pathway

作　　者：王生华 朱莹 王霈 吴珊 李擎虎[1] WANG Sheng-hua;ZHU Ying;WANG Yi-pei;WU Shan;LI Qing-hu(Department of Spleen and Gastric Diseases,Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,410005;Department of Pulmonary Spleen and Gastric Diseases,Loudi Hospital of Traditional Chinese Medicine,Hunan,417000;First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha,410007)

机构地区：[1]湖南中医药大学第二附属医院消化内科,长沙410005 [2]湖南省娄底市中医医院肺脾胃病科,湖南417000 [3]湖南中医药大学第一附属医院,长沙410007

出　　处：《中国中西医结合杂志》2021年第8期951-958,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：湖南省中医药科研计划项目一般课题(No.201942,No.2020043)。

摘　　要：目的基于Nrf2信号通路探讨溃结宁膏穴位敷贴在脾肾阳虚型溃疡性结肠炎(UC)大鼠肠黏膜抗氧化应激的作用机制。方法将65只雄性SD大鼠,随机分为空白组(10只)、造模组(55只),造模组采用腺嘌呤、冰番泻叶分阶段灌胃结合外因干预,并予2,4,6-三硝基苯磺酸与乙醇复合物灌肠制备UC病证结合动物模型,模型评价后将造模组大鼠按照体重随机分为模型组(8只)、穴位组(7只)、非穴位组(7只)、柳氮磺胺吡啶(SASP)组(8只)、SASP合穴位组(8只)。病证结合模型大鼠分别予溃结宁膏穴位敷贴"中脘""气海""足三里""天枢",溃结宁膏非穴位敷贴于双臀部肌肉丰厚处,SASP组予SASP灌胃,SASP合穴位组予穴位敷贴合SASP灌胃同步处理,每日1次,共28天。观察大鼠疾病活动指数(DAI)评分、结肠组织病理改变、结肠黏膜损伤指数(CMDI)评分、结肠组织学损伤指数(TDI)评分,同时运用免疫组化法检测结肠组织中Nrf2和HO-1蛋白表达,PCR检测Nrf2、Keap1 mRNA表达。结果与空白组比较,模型组CMDI、TDI评分、Nrf2和HO-1蛋白含量及Nrf2和Keap1 mRNA表达升高(P<0.05,P<0.01)。与模型组比较,穴位组、SASP组、SASP合穴位组CMDI、TDI评分降低(P<0.01),Nrf2、HO-1蛋白含量及Nrf2 mRNA表达升高(P<0.01)而Keap1mRNA表达降低(P<0.05,P<0.01);与SASP组比较,穴位组、SASP合穴位组各项指标比较差异无统计学意义(P>0.05)。结论溃结宁膏穴位敷贴通过调控氧化应激反应中的Nrf2信号通路,上调Nrf2、HO-1蛋白和Nrf2 mRNA的表达,并下调Keap1 mRNA的表达,抵御结肠氧化应激反应,减轻肠黏膜炎症,促进结肠黏膜愈合,进而发挥治疗UC的作用。Objective To study the mechanism of anti-oxidative stress in intestinal mucosa of ulcerative colitis(UC) rats with Pi-Shen yang deficiency syndrome(PSYDS) based on Nrf2 signaling pathway. Methods Sixty-five male SD rats were randomly divided into blank group(n=10) and model group(n= 55). The rat model was established by gavage of the decoction of Folium Sennae and adenine at different stages combined with external cause intervention, as well as enema with the compound of 2,4, 6-trinitrobenzene sulfonic acid and alcohol. After model evaluation, rats in the model group were randomly divided into model group(n=8), acupoint group(n=7), non-acupoint group(n=7), salazosulfapyridine(SASP) group(n=8), and SASP combined acupoint group(n =8) according to weight. The acupoint application group was given Kuijienning Plaster(KJNP) on acupoint application(Zhongwan,Qihai,Zusanli, Tianshu). Non-acupoint application group was given KJNP to the rich muscles of both buttocks. SASP group was given intragastric administration of SASP,while the SASP combined acupoint group was given SASP combined with acupoint application at the same time, once a day, for a total of 28 days. The general condition,disease activity index(DAI) score, pathological changes of colon tissue,colonic mucosal injury index(CMDI) score and tissue damage index(TDI) score were observed. Meanwhile, the expressios of Nrf2 and HO-1 protein in colon tissue were detected by immunohistochemical method. Nrf2 and KEAP1 mRNA expressions were detected by PCR.Results Compared with blank group,CMDI and TDI scores,the protein contents of Nrf2 and HO-1,and mRNA expressios of Nrf2 and Keap1 in model group increased(P<0.05, P<0.01). Compared with model group, the CMDI and TDI scores in acupoint group, SASP group and SASP combined acupoint group decreased(P<0.01). The protein contents of Nrf2 and HO-1 and mRNA expression of Nrf2 increased(P<0.01), while the mRNA expression of KEAP1 decreased(P<0.05,P<0.01). Compared with SASP group, there was no statistical significance in each ind

关 键 词：溃疡性结肠炎 脾肾阳虚证 溃结宁膏 Nrf2 HO-1 KEAP1 

分 类 号：R285.5[医药卫生—中药学]