摘要
目的探讨25-羟基胆固醇(25-HC)在小鼠炎症性肠病(IBD)发生发展中的作用及其机制。方法全部小鼠分为三组:对照组正常喂养;DSS组用2.5%葡聚糖硫酸钠(DSS)溶液喂养;DSS+25-HC组用2.5%DSS溶液喂养,且腹腔注射25-HC溶液。通过评估小鼠的疾病活动指数判断小鼠的症状变化,通过组织学评分判断小鼠的组织改变并利用Western blot、qRT-PCR、免疫组化、流式细胞术等检测小鼠白细胞介素-17(IL-17)的表达情况及信号通路,并结合小鼠肠上皮紧密连接蛋白的检测,探究25-HC影响小鼠IBD的机制。结果与DSS组比较,DSS+25-HC组小鼠体质量减轻(F=30.1,P<0.0001),结肠缩短(F=63.8,P<0.05),疾病活动指数(F=774.5,P<0.0001)和组织病理学评分升高(F=141.5,P<0.05),紧密连接相关蛋白(ZO-2、Occludin、JAM和Claudin-4)表达显著降低,IL-17水平显著降低,且表达水平与紧密连接蛋白呈正相关。结论25-HC通过RORγt通路抑制结肠γδT细胞IL-17的产生,加重黏膜损伤,促进小鼠IBD发展。
Objective To explore the role and mechanism of 25-hydroxycholesterol(25-HC)in inflammatory bowel disease(IBD)in mice.Methods All mice were divided into three groups:the control group was fed normally;the DSS model group was fed with 2.5%dextran sulfate sodium(DSS)solution;the DSS+25-HC experimental group was fed with 2.5%DSS solution and he mice in the experimental group were intraperitoneally injected with 25-HC.The symptom changes of the mice were evaluated by assessing the disease activity index(DAI),and the tissue changes were judged by histological scoring.The expression of interleukin-17 and its signaling pathways in the mice were detected by Western blot,qRT-PCR,immunohistochemistry/fluorescence,and flow cytometry.Combined with the detection of tight junction proteins in the intestinal epithelium of the mice,the mechanism by which 25-HC affects IBD in mice was explored.Results In comparison to the DSS control group,The DSS+25-HC experimental group mice exhibited a reduction in body weight(F=30.1,P<0.0001),a shortened colon(F=63.8,P<0.05),and elevated DAI(F=774.5,P<0.0001)and histopathological scores(F=141.5,P<0.05).Additionally,the expression of tight junction-associated proteins(ZO-2,Occludin,JAM and Claudin-4)was found to be significantly reduced.The level of IL-17 significantly decreased,and its expression level was positively correlated with tight junction proteins.Conclusion 25-HC inhibited IL-17 production by colonicγδT cells through the RORγt pathway,aggravated mucosal injury,and promoted the development of DSS-induced acute colitis in mice.
作者
李雨桐
罗小琦
谭启发
陈明杰
吴长有
沈娟
Li Yutong;Luo Xiaoqi;Tan Qifa;Chen Mingjie;Wu Changyou;Shen Juan(KingMed School of Laboratory Medicine,Guangzhou Medical University,Guangzhou 511436;Guangzhou Key Laboratory for Clinical Rapid Diagnosis and Early Warning of Infectious Diseases,Guangzhou 511436;Engineering Technology Research Center of Intelligent Diagnosis for Infectious Diseases in Guangdong Province,Guangzhou 511436;Dept of Laboratory Medicine,The Third People′s Hospital of Ganzhou City,Ganzhou 341000;Institute of Immunology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 511400)
出处
《安徽医科大学学报》
北大核心
2025年第7期1204-1212,共9页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:82001747)
广东省基础与应用基础研究基金(编号:2020A1515110562)。
