艾灸調節潰瘍性結腸炎小鼠E3泛素連接酶TRIM31的腸黏膜屏障保護機制

吳璐一 鐘蕊 林亞瑩 鄭寒丹 劉世敏 黃艷 馬喆 劉雅楠

摘要 目的：觀察艾灸對野生型和TRIM31基因敲除（TRIM31-/-）潰瘍性結腸炎（UC）模型小鼠緊密連接蛋白以及血清炎癥介質的影響，探討TRIM31能否通過保護UC腸黏膜屏障而降低UC腸道炎癥反應以及艾灸對其調節作用。方法：分別采用野生型和TRIM31基因敲除小鼠構建UC模型，免疫組織化學檢測結腸組織中TRIM31、閉合蛋白、密封蛋白、ZO-1的蛋白表達;實時熒光定量PCR（RT-qPCR）檢測法檢測結腸組織中TRIM31的mRNA表達;ELISA檢測血清白細胞介素-1β（IL-1β）、IL-13、IL-25、IL-33蛋白的濃度。結果：艾灸可改善野生型UC小鼠的結腸炎癥，升高結腸TRIM31蛋白及mRNA的表達以及腸黏膜屏障相關閉合蛋白、密封蛋白、ZO-1的表達，降低血清炎癥介質IL-1β、IL-13、IL-25、IL-33的濃度;TRIM31-/-小鼠較野生型小鼠相比，結腸炎癥加重，腸黏膜屏障相關閉合蛋白、密封蛋白、ZO-1表達降低，血清炎癥介質IL-1β、IL-13濃度升高;艾灸還可改善TRIM31-/-UC小鼠的結腸炎癥，升高結腸組織腸黏膜屏障相關閉合蛋白、密封蛋白、ZO-1的表達，降低血清炎癥介質IL-1β、IL-13、IL-25、IL-33的濃度。結論：TRIM31蛋白可能是艾灸參與調節UC腸黏膜屏障保護的關鍵靶點;艾灸對TRIM31-/-小鼠腸黏膜屏障相關閉合蛋白、密封蛋白、ZO-1及血清炎癥介質IL-1β、IL-13、IL-25、IL-33仍有調節作用，表明艾灸還可能通過發揮其多靶點調節作用參與UC腸黏膜屏障保護。

關鍵詞 潰瘍性結腸炎;艾灸;TRIM31;腸黏膜屏障;炎癥介質;基因敲除

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

WU Luyi1，2，ZHONG Rui3，LIN Yaying2，ZHENG Handan2，LIU Shimin2，HUANG Yan2，MA Zhe2，LIU Yanan2

（1 Yueyang Hospital of Integrated Traditional Chinese and Western Medicine，Shanghai University of Traditional Chinese Medicine，Shanghai 200437，China; 2 Shanghai Research Institute of Acupuncture and Meridian，Shanghai 200030，China; 3 Shanghai Qigong Research Institute，Shanghai University of Traditional Chinese Medicine，Shanghai 200030，China）

Abstract 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-1β and 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.

Keywords Ulcerative colitis; Moxibustion; TRIM31; Intestinal mucosal barrier; Inflammatory factors; Gene knockout

中圖分類號：R256.3文獻標識碼：Adoi：10.3969/j.issn.1673-7202.2022.03.007

潰瘍性結腸炎（Ulcerative Colitis，UC）是一種病因未明的慢性復發性結直腸疾病，與克羅恩?。–rohn Disease，CD）同屬于炎癥性腸病的范疇。UC的臨床癥狀主要包括腹瀉、腹痛、黏液膿血便、體質量減輕、里急后重等，另外還可表現為皮膚、眼睛、關節和肝膽等腸外癥狀。……