紫菀酮通过抑制肠上皮细胞凋亡改善TNBS诱导的克罗恩病样结肠炎  

Shionone improves TNBS-induced Crohn′s disease like colitis by inhibiting apoptosis of intestinal epithelial cells

作　　者：徐梦宇 杨子 张文静 张小凤 XU Mengyu;YANG Zi;ZHANG Wenjing;ZHANG Xiaofeng(Central Laboratory,First Affiliated Hospital of Bengbu Medical University,Bengbu 233004,China;Anhui Province Key Laboratory of Basic and Translational Research of Inflammation-Related Diseases;Department of Gastrointestinal Surgery,First Affiliated Hospital of Bengbu Medical University;Clinical Laboratory,First Affiliated Hospital of Bengbu Medical University)

机构地区：[1]蚌埠医科大学第一附属医院中心实验室,蚌埠233004 [2]炎症相关性疾病基础与转化研究安徽省重点实验室 [3]蚌埠医科大学第一附属医院胃肠外科 [4]蚌埠医科大学第一附属医院检验科

出　　处：《山西医科大学学报》2024年第3期319-325,共7页Journal of Shanxi Medical University

基　　金：蚌埠医学院自然科学基金重点项目(2022byzd073)。

摘　　要：目的研究紫菀酮(SHI)对2,4,6-三硝基苯磺酸(TNBS)诱导的小鼠克罗恩病(CD)样结肠炎的作用及潜在机制。方法18只野生型小鼠随机分为3组:对照组(WT组)、模型组(TNBS组)和SHI干预组(SHI组),每组6只。TNBS组小鼠肛门推注5%TNBS诱导CD样结肠炎;SHI组在TNBS造模的同时给予SHI灌胃干预[40 mg/(kg·d)],WT组和TNBS组每日给予等量的生理盐水灌胃。1周后处死小鼠,采用疾病活动度(DAI)评分、体质量改变、结肠长度、组织学炎症评分和小鼠结肠黏膜肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平评估肠炎程度。采用荧光素异硫氰酸酯-右旋糖酐(FITC)渗透性测定、肠型脂肪酸结合蛋白(I-FABP)测定、细菌移位率评估肠屏障功能。通过TUNEL染色检测小鼠肠上皮细胞凋亡情况;通过Western blot检测小鼠凋亡相关蛋白(Bcl-2和Bax)的表达量。采用网络药理学以及生信富集分析预测SHI干预对小鼠CD样结肠炎保护作用的可能途径。结果与WT组比较,TNBS组DAI评分、体质量降低幅度、组织学炎症评分和结肠黏膜TNF-α、IL-6、IL-1β水平显著升高(P<0.05);与TNBS组比较,SHI组DAI评分、体质量降低幅度、组织学炎症评分和结肠黏膜TNF-α、IL-6、IL-1β水平显著降低(P<0.05)。与WT组比较,TNBS组小鼠结肠长度缩短(P<0.05);与TNBS组比较,SHI组小鼠结肠长度增加(P<0.05)。网络药理学以及生信富集分析发现SHI可能通过拮抗肠上皮细胞凋亡治疗CD。与TNBS组比较,SHI组小鼠外周血FITC-dextran、I-FABP水平和肝脏、肠系膜淋巴结的细菌移位率较低(P<0.05)。TUNEL染色结果显示,与TNBS组比较,SHI组小鼠结肠组织中肠上皮细胞凋亡数量显著减少(P<0.05),Bax的表达下调,Bcl-2的表达上调(均P<0.05)。结论SHI可能通过抑制肠上皮细胞凋亡进而缓解TNBS诱导的小鼠CD样结肠炎。Objective To investigate the effect and its possible mechanism of shionone(SHI)on Crohn′s disease(CD)-like colitis induced by 2,4,6-trinitrobenzene sulfonic acid(TNBS)in mice.Methods Eighteen wild-type mice were randomly divided into three groups:control group(WT group),model group(TNBS group),and SHI intervention group(SHI group),with 6 mice in each group.Mice in TNBS group were given 5%TNBS by anal injection to induce CD-like colitis,while mice in SHI group were given SHI[40 mg/(kg·d)]by gavage at the same time of TNBS modeling,and mice in WT group and TNBS group were given equal volume of normal saline by gavage daily.One week later,the mice were euthanized and the severity of colitis was evaluated by disease activity(DAI)score,body mass change,colon length,histological inflammation score,and the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-1β(IL-1β)in mouse colon mucosa.Intestinal barrier function was evaluated by fluorescein isothiocyanate-dextran(FITC)permeability,intestinal fatty acid-binding protein(I-FABP)and bacterial displacement rate.The apoptosis of intestinal epithelial cells in each group was detected by TUNEL staining.The expressions of apoptosis-related proteins(Bcl-2 and Bax)in each group were detected by Western blot.The protective effect of SHI intervention against CD-like colitis in mice was predicted by network pharmacology and biogenic enrichment analysis.Results Compared with WT group,DAI score,body mass reduction,histological inflammation score and the levels of TNF-α,IL-6 and IL-1βin colon mucosa were significantly increased in TNBS group(P<0.05).Compared with TNBS group,DAI score,body mass reduction,histological inflammation score and the levels of TNF-α,IL-6 and IL-1βin colon mucosa were significantly decreased in SHI group(P<0.05).Compared with WT group,the colonic length was shortened in TNBS group(P<0.05).Compared with TNBS group,the colonic length was increased in SHI group(P<0.05).Network pharmacology and biogenic enrichment analysis showe

关 键 词：炎症性肠病 克罗恩病 紫菀酮 肠上皮细胞凋亡 肠道炎症 

分 类 号：R574.62[医药卫生—消化系统]