麦粒灸调控TLR4/MyD88/NF-κB信号通路减轻溃疡性结肠炎小鼠结肠损伤的机制研究  

Moxibustion with seed-size moxa cones alleviates colonic injury in mice with ulcerative colitis by regulating TLR4/MyD88/NF-κB signaling pathway

作　　者：朱涛 冀来喜 任佳 程艳婷 冀雨芳 高珍 张静 王海军 ZHU Tao;JI Lai-xi;REN Jia;CHENG Yan-ting;JI Yu-fang;GAO Zhen;ZHANG Jing;WANG Hai-jun(College of Acupuncture-moxibustion and Tuina,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China;The Second Clinical College,Shanxi University of Chinese Medicine,Jinzhong 030619,Shanxi Province;Affiliated Hospital of Shanxi University of Chinese Medicine,Taiyuan 030024)

机构地区：[1]成都中医药大学针灸推拿学院,成都611137 [2]山西中医药大学第二临床学院,山西晋中030619 [3]山西中医药大学附属医院,太原030024

出　　处：《针刺研究》2024年第8期836-844,共9页Acupuncture Research

基　　金：国家自然科学基金项目(No.82074549)。

摘　　要：目的:观察麦粒灸对溃疡性结肠炎(UC)小鼠Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核转录因子κB(NF-κB)信号通路的影响,探讨麦粒灸对UC结肠损伤的治疗机制。方法:C57BL/6雄性小鼠随机分为空白组、模型组、麦粒灸组、西药组,每组10只。采用连续7 d自由饮用3%DSS溶液法制备UC小鼠模型。麦粒灸组予麦粒灸“中脘”“天枢”“上巨虚”治疗,每穴3壮,每壮时间约为30 s。西药组给予300 mg/kg美沙拉嗪肠溶片溶液灌胃。各组干预均1次/d,连续7 d。每隔2 d观察小鼠一般情况,计算小鼠疾病活动指数(DAI)评分;HE染色法观察小鼠结肠组织形态;ELISA法检测血清白细胞介素(IL)-1β、肿瘤坏死因子α(TNF-α)、IL-6和IL-8的含量;免疫组织化学法检测小鼠结肠组织中TLR4、MyD88的阳性表达;实时荧光定量PCR法检测小鼠结肠组织中TLR4、MyD88、NF-κB p65 mRNA的表达水平。结果:与空白组比较,模型组小鼠出现不同程度的软便或稀便,结肠长度缩短(P<0.01)、体质量下降(P<0.01),DAI评分、肠重指数、黏膜损伤评分、结肠病理评分及血清IL-1β、TNF-α、IL-6和IL-8含量升高(P<0.01),结肠组织中TLR4、MyD88阳性表达及TLR4、MyD88、NF-κB p65 mRNA的表达均升高(P<0.01);与模型组比较,麦粒灸组、西药组小鼠粪便性状得到了改善,结肠长度增加(P<0.01)、体质量升高(P<0.01),DAI评分、肠重指数、黏膜损伤评分、结肠病理评分及血清IL-1β、TNF-α、IL-6和IL-8含量降低(P<0.01,P<0.05),结肠组织中TLR4、MyD88阳性表达及TLR4、MyD88、NF-κB p65 mRNA的表达均下降(P<0.05,P<0.01);麦粒灸与西药组相比,以上指标差异无统计学意义。结论:麦粒灸可能通过调控TLR4/MyD88/NF-κB信号通路,减少炎性因子的释放,从而改善UC小鼠的结肠损伤。Objective To observe the effect of moxibustion with seed-size moxa cones on the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factor-κB(NF-κB)signaling pathway in mice with ulcerative colitis(UC),so as to explore the therapeutic mechanism of moxibustion with seed-size moxa cones on colonic injury in UC.Methods Forty male C57BL/6 mice were randomly divided into blank group,model group,moxibustion group,and western medicine group,with 10 mice in each group.The UC mouse model was established by 3%DSS solution by free drinking for 7 consecutive days.Mice in the moxibustion group were treated with seed-size moxa cones at“Zhongwan”(CV12),“Tianshu”(ST25)and“Shangjuxu”(ST37),3 moxa cones per point,with each cone applied for approximately 30 s,while mice in the western medicine group were orally administered with 300 mg/kg mesalazine solution,which were both conducted once a day for 7 consecutive days.The general condition of mice was observed every 2 days,and the disease activity index(DAI)score was calculated.HE staining was used to observe the morphology of colonic tissue in mice.ELISA was used to detect the serum interleukin(IL)-1β,tumor necrosis factor(TNF)-α,IL-6,and IL-8 contents.Immunohistochemistry was used to detect the positive expression of TLR4 and MyD88 in colonic tissue of mice.Real-time fluorescence quantitative PCR was used to detect the expression levels of TLR4,MyD88,and NF-κB p65 mRNAs in colonic tissue.Results Compared with the blank group,varying degrees of soft or watery stools were observed,colon length and body weight were decreased(P<0.01)in mice of the model group,while DAI score,colon weight index,mucosal damage score,colonic pathological score,serum IL-1β,TNF-α,IL-6,and IL-8 contents,positive expressions of TLR4 and MyD88,and TLR4,MyD88,and NF-κB p65 mRNA expressions in colonic tissue were increased(P<0.01).Compared with the model group,improved fecal characteristics were observed,colon length and body weight were increased(P<0.01

关 键 词：麦粒灸 溃疡性结肠炎 结肠损伤 炎性反应 TLR4/MyD88/NF-κB信号通路 

分 类 号：R245[医药卫生—针灸推拿学]