基于TLR4/MyD88/NF-κB信号通路和肠黏膜屏障保护因子探讨魔芋润肠复合膳食纤维治疗便秘大鼠作用机制  

Efficacy and Mechanism of Konjac Runchang(润肠)Compound Dietary Fiber against Constipation in Rats:Based on TLR4/My D88/NF-κB Signaling Pathway and Intestinal Mucosal Barrier Protective Factor

作　　者：廖勇 贺海波[2] 李洁[2] 王晓 游艳[1] 郭阳 汤枝鹏 李璐[1] 张继红[3] 罗涛[3] Liao Yong;He Haibo;Li Jie;Wang Xiao;You Yan;Guo Yang;Tang Zhipeng;Li Lu;Zhang Jihong;Luo Tao(Department of Pharmacy,Afiliated Renhe Hospital of China Three Gorges University,Yichang443002;Hubei Key Laboratory of Natural Products Research and Development,Yichang Key Laboratory of Development and Utilization of Health Products with Drug and Food Homology,China Three Gorges University,Yichang 443002;Chinese Medicine Clinical Medical College of China Three Gorges University&Hubei Clinical Research Center for Functional Digestive Diseases of Traditional Chinese Medicine,Yichang 443002)

机构地区：[1]三峡大学附属仁和医院药学部,宜昌昌443002 [2]三峡大学天然产物研究与利用湖北省重点实验室,药食同源大健康产品开发利用宜昌市重点实验室,宜昌443002 [3]三峡大学中医临床医学院&湖北省功能性消化系统疾病中医临床医学研究中心,宜昌443002

出　　处：《中药药理与临床》2022年第6期75-83,共9页Pharmacology and Clinics of Chinese Materia Medica

基　　金：湖北省卫生健康委员会中医药科研项目(编号:ZY2019M074);宜昌市科学技术局项目(编号:A21-2-044);三峡大学硕士学位论文培优基金项目(编号:2020SSPY144、2019SSPY159);湖北省生物酵素工程研究技术研究中心项目(编号:JS2018-06)。

摘　　要：目的:探讨魔芋润肠复合膳食纤维对便秘大鼠的治疗作用及对Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子-κB(NF-κB)炎症通路和结肠黏膜屏障的影响。方法:将SD大鼠随机分为正常对照组、模型对照组、魔芋润肠复合膳食纤维0.54、1.08 g/kg组和首荟通便胶囊0.38 g/kg组。大鼠2次/d腹腔注射洛哌丁胺3 mg/kg,第8 d后,各给药组灌胃给予相应药物。给药2 w后,进行体质量、食物摄入量、首次排便时间、排便粒数/质量、粪便含水量、胃排空率、肠道推进率和粪便短链脂肪酸测定;测定血液中白细胞介素(IL-4)、IL-10和肿瘤坏死因子-α(TNF-α)、IL-1β、IL-6、γ干扰素(IFN-γ)、胃泌素(GAS)、胃动素(MTL)、生长抑素(SS)、P物质(SP)、血管活性肽(VIP)和乙酰胆碱酯酶(ACh E)、降钙素基因相关肽(CGRP)、ROS、LDH、GSH-Px、SOD、CAT、MDA和MPO含量或活力,观察结肠组织形态学、病理学评分;Real-time PCR检测结肠组织中Tlr4、Myd88、Nfκb、Claudin1、E-cadherin、Occludin、Muc2、Tff3、Zo1 mRNA表达;Western blot检测结肠组织中Claudin1、E-cadherin、Occludin、MUC-2、TFF3、ZO-1蛋白表达。结果:正常对照组比较,模型对照组大鼠体质量、摄食量和结肠病理学评分明显升高,结肠黏膜完整性恶化,排便时间延长,胃排空率、小肠推进率及24 h排泄粪便粒数、粪便湿、干质量和含水量显著降低(P<0.01);血清中IFN-γ、IL-1β、IL-6、TNF-α、SS、VIP、CGRP、LDH、ROS、MDA活力或含量显著升高,结肠组织中MPO含量显著升高(P<0.01);血清中IL-4、IL-10、SOD、CAT、GSH-Px、Gas、MTL、SP、AChE含量显著降低,粪便中总SCFA和乙酸、丙酸、丁酸含量降低(P<0.05或P<0.01);结肠组织中Tlr4、Myd88、Nfκb mRNA表达水平上调,结肠组织中黏膜屏障保护因子Claudin1、E-cadherin、Occludin、Muc2、Tff3、Zo1 mRNA和蛋白表达显著下调(P<0.01);用魔芋润肠复合膳食纤维0.54、1.08 g/kg组治疗后,上述指�Objective:To investigate the therapeutic effect of Konjac Runchang compound dietary fiber(KRCDF)on constipation rats,the toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB)signaling pathway,and colonic mucosal barrier.Methods:SD rats were randomized into control group,model group,KRCDF(0.54 and 1.08 g/kg)groups,and Shouhui Tongbian Capsules(首荟通便胶囊)(STC,0.38 g/kg)group.The experimental rats were injected(ip)with loperamide(3 mg/kg,twice/day)to induce the constipation.After 8 days,administration(ig)began and lasted 2 weeks.Then the body weight,food intake,time of the first defecation,number/weight of feces,fecal water content,gastric emptying rate,intestinal propulsion rate,and fecal short chain fatty acids were detected,and the blood interleukin(IL)-4,IL-10,tumor necrosis factor-α(TNF-α),IL-1β,IL-6,interferon-γ(IFN-γ),gastrin(Gas),motilin(MTL),somatostatin(SS),substance P(SP),vasoactive peptide(VIP),acetylcholinesterase(AChE),calcitonin gene-related peptide(CGRP),reactive oxygen species(ROS),lactate dehydrogenase(LDH),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),catalase(CAT),malondialdehyde(MDA),and myeloperoxidase(MPO)were detected.Colon histomorphology was observed and pathological scores were recorded.Real-time PCR was applied to test the mRNA expression of Tlr4,Myd88,Nfκb,Claudin1,E-cadherin,Occludin,Muc2,Tff3,and Zo1,and Western blotting to determine the protein expression of Claudin1,E-cadherin,Occludin,MUC-2,TFF3,and ZO-1 in the colon tissue.Results:Compared with the control group,the model group had high body weight,food intake,and colonic pathological score,severe colonic mucosal edema and inflammatory infiltration,long defecation time,low gastric emptying rate,intestinal propulsion rate,number of feces excreted in 24 h,wet weight of feces,dry weight of feces,and fecal water content(P<0.01),high serum content/activity of IFN-γ,IL-1β,IL-6,TNF-α,SS,VIP,CGRP,LDH,ROS,and MDA,and MPO activity in colon tissue(P<0.01),low serum content

关 键 词：魔芋润肠复合膳食纤维 便秘 氧化应激 炎症反应 胃肠激素 肠道屏障 Toll样受体4/髓样分化因子88/核转录因子-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]