艾灸调节溃疡性结肠炎小鼠E3泛素连接酶TRIM31的肠黏膜屏障保护机制  

Protective Mechanism of Moxibustion on Intestinal Mucosal Barrier by Regulating E3 Ubiquitin Ligase TRIM31 in Mice with Ulcerative Colitis

作　　者：吴璐一[1,2] 钟蕊 林亚莹 郑寒丹 刘世敏[2] 黄艳[2] 马喆[2] 刘雅楠[2] WU Luyi;ZHONG Rui;LIN Yaying;ZHENG Handan;LIU Shimin;HUANG Yan;MA Zhe;LIU Yanan(Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Shanghai Research Institute of Acupuncture and Meridian,Shanghai 200030,China;Shanghai Qigong Research Institute,Shanghai University of Traditional Chinese Medicine,Shanghai 200030,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院,上海200437 [2]上海市针灸经络研究所,上海200030 [3]上海中医药大学上海市气功研究所,上海200030

出　　处：《世界中医药》2022年第3期329-335,共7页World Chinese Medicine

基　　金：国家自然科学基金项目(81973955,82004475);上海市卫生计生系统优秀人才培养计划-优秀青年人才(2018YQ11);上海市卫生健康委员会科研课题计划资助(20194Y0005);上海市重点专科(针灸)建设(shslczdzk04701)。

摘　　要：目的:观察艾灸对野生型和TRIM31基因敲除(TRIM31^(-/-))溃疡性结肠炎(UC)模型小鼠紧密连接蛋白以及血清炎症介质的影响,探讨TRIM31能否通过保护UC肠黏膜屏障而降低UC肠道炎症反应以及艾灸对其调节作用。方法:分别采用野生型和TRIM31基因敲除小鼠构建UC模型,免疫组织化学检测结肠组织中TRIM31、闭合蛋白、密封蛋白、ZO-1的蛋白表达;实时荧光定量PCR(RT-qPCR)检测法检测结肠组织中TRIM31的mRNA表达;ELISA检测血清白细胞介素-1β(IL-1β)、IL-13、IL-25、IL-33蛋白的浓度。结果:艾灸可改善野生型UC小鼠的结肠炎症,升高结肠TRIM31蛋白及mRNA的表达以及肠黏膜屏障相关闭合蛋白、密封蛋白、ZO-1的表达,降低血清炎症介质IL-1β、IL-13、IL-25、IL-33的浓度;TRIM31^(-/-)小鼠较野生型小鼠相比,结肠炎症加重,肠黏膜屏障相关闭合蛋白、密封蛋白、ZO-1表达降低,血清炎症介质IL-1β、IL-13浓度升高;艾灸还可改善TRIM31^(-/-)UC小鼠的结肠炎症,升高结肠组织肠黏膜屏障相关闭合蛋白、密封蛋白、ZO-1的表达,降低血清炎症介质IL-1β、IL-13、IL-25、IL-33的浓度。结论:TRIM31蛋白可能是艾灸参与调节UC肠黏膜屏障保护的关键靶点;艾灸对TRIM31^(-/-)小鼠肠黏膜屏障相关闭合蛋白、密封蛋白、ZO-1及血清炎症介质IL-1β、IL-13、IL-25、IL-33仍有调节作用,表明艾灸还可能通过发挥其多靶点调节作用参与UC肠黏膜屏障保护。Objective:To observe the effects of moxibustion on tight junction proteins and serum inflammatory factors in wild-type and TRIM31 knockout(TRIM31^(-/-))ulcerative colitis(UC)model mice and explore the possibility of TRIM31 in reducing intestinal inflammation in UC by protecting the intestinal mucosal barrier,as well as the regulatory role of moxibustion.Methods:The UC model was induced in the wild-type and TRIM31^(-/-)mice.Immunohistochemistry was used to detect the protein expression of TRIM31,Occludin,Claudin-1,and ZO-1 in colon tissues.RT-qPCR was used to detect the mRNA expression of TRIM31 in colon tissues.The serum levels of IL-1β,IL-13,IL-25,and IL-33 were detected by ELISA.Results:Moxibustion could resist colonic inflammation in wild-type UC mice,increase the protein and mRNA expression of TRIM31 and the expression of Occludin,Claudin-1,and ZO-1 in the colon,and reduce the concentrations of inflammatory factors IL-1β,IL-13,IL-25,and IL-33 in the serum.Compared with the wild-type mice,the TRIM31^(-/-)mice showed aggravated colitis,decreased expression of Occludin,Claudin-1,and ZO-1,and increased serum concentrations of IL-1βand IL-13.Additionally,moxibustion could improve the colitis of TRIM31^(-/-)UC mice,increase the expression of Occludin,Claudin-1,and ZO-1 in the colon,and reduce the serum concentrations of IL-1β,IL-13,IL-25,and IL-33.Conclusion:TRIM31 protein might serve as a key target of moxibustion in regulating the intestinal mucosal barrier in UC.The regulatory role of moxibustion on Occludin,Claudin-1,ZO-1,IL-1β,IL-13,IL-25,and IL-33 in TRIM31^(-/-)mice was verified,which suggested that moxibustion presumedly participated in the protection of intestinal mucosal barrier in UC by virtue of its multi-target regulatory role.

关 键 词：溃疡性结肠炎 艾灸 TRIM31 肠黏膜屏障 炎症介质 基因敲除 

分 类 号：R256.3[医药卫生—中医内科学]