紫花前胡苷通过抑制肠上皮细胞焦亡改善2,4,6-三硝基苯磺酸诱导的小鼠实验性结肠炎  

Nodakenin ameliorates TNBS-induced experimental colitis in mice by inhibiting pyroptosis of intestinal epithelial cells

作　　者：黄菊 殷丽霞 牛民主 耿志军 左芦根 李静 胡建国 HUANG Ju;YIN Lixia;NIU Minzhu;GENG Zhijun;ZUO Lugen;LI Jing;HU Jianguo(Central Laboratory,First Affiliated Hospital of Bengbu Medical University,Bengbu 233003,China;Anhui Provincial Key Laboratory of Basic and Translational Research of Inflammation-related Diseases,First Affiliated Hospital of Bengbu Medical University,Bengbu 233003,China;Department of Clinical Laboratory,First Affiliated Hospital of Bengbu Medical University,Bengbu 233003,China;Gastrointestinal Surgery,First Affiliated Hospital of Bengbu Medical University,Bengbu 233003,China;Department of Immunology,School of Laboratory Medicine,Bengbu Medical University,Bengbu 233000,China)

机构地区：[1]蚌埠医科大学第一附属医院中心实验室,安徽蚌埠233003 [2]蚌埠医科大学第一附属医院炎症相关性疾病基础与转化研究安徽省重点实验室,安徽蚌埠233003 [3]蚌埠医科大学第一附属医院检验科,安徽蚌埠233003 [4]蚌埠医科大学第一附属医院胃肠外科,安徽蚌埠233003 [5]蚌埠医科大学检验医学院免疫学教研室,安徽蚌埠233000

出　　处：《南方医科大学学报》2025年第2期261-268,共8页Journal of Southern Medical University

基　　金：国家自然科学基金(82370534,82070561);安徽省高校优秀青年基金项目(2022AH030138);安徽省教育厅优秀青年教师培育项目(YQYB2023019);蚌埠医科大学自然科学重大科技项目孵育计划(2020byfy003);蚌埠医科大学国家自然科学基金面上项目孵育项目(2023byfy002)。

摘　　要：目的探讨天然植物单体紫花前胡苷(Nod)对克罗恩病(CD)样结肠炎的影响及其作用机制。方法建立脂多糖和三磷酸腺苷(ATP)联合诱导的结肠类器官焦亡模型,通过检测焦亡关键调节因子、通透性和促炎因子,探讨Nod对细胞焦亡、肠道屏障功能和炎症反应的影响。以2,4,6-三硝基苯磺酸(TNBS)干预小鼠为CD动物模型,通过检测体质量、DAI评分、组织病理学分析、炎症评分、肠屏障功能和肠上皮细胞焦亡,探讨Nod对CD样结肠炎的治疗效果。通过网络药理学和体内、体外实验验证探索Nod保护肠上皮细胞焦亡的潜在机制。结果脂多糖和ATP诱导的结肠类器官经Nod干预后显著抑制NLRP3、GSDMD-N、cleaved caspase-1和caspase-11的表达,改善肠道FITC-dextran(FD4,4000)通透性,以及降低IL-1β和IL-18水平(P<0.05)。在TNBS诱导的结肠炎小鼠中,Nod治疗后能缓解小鼠体质量下降幅度、降低DAI评分、改善炎症细胞浸润和炎症评分(P<0.05),并降低血清中FD4、I-FABP的含量与细菌移位至肠系膜淋巴结、脾和肝脏中比例(P<0.05)。Nod可抑制小鼠肠黏膜中NLRP3、GSDMD-N、cleaved caspase-1和caspase-11的表达(P<0.05)。网络药理学预测分析显示,Nod抗结肠炎可能与PI3K/Akt通路有关;体内外实验证实,Nod抑制PI3K/Akt通路的激活,且PI3K/Akt通路的激活剂(IGF-1)逆转了Nod对肠上皮细胞焦亡和肠屏障功能的保护作用(P<0.05)。结论Nod至少部分通过抑制PI3K/Akt信号传导拮抗肠上皮细胞的焦亡,从而保护肠屏障功能和改善CD样结肠炎,有望成为一种新的CD治疗药物。Objective To investigate the therapeutic mechanism of nodakenin for Crohn's disease(CD)-like colitis in mice.Methods Using a colonic organoid model with lipopolysaccharide(LPS)-and ATP-induced pyroptosis,we investigated the effects of nodakenin on pyroptosis,intestinal barrier function and inflammatory response by detecting key pyroptosis-regulating factors and assessing changes in permeability and pro-inflammatory factors.In a mouse model of 2,4,6-trinitrobenzenesulfonic acid(TNBS)-induced CD-like colitis,the therapeutic effect of nodakenin was evaluated by measuring changes in body weight,DAI score,colonic histopathologies,inflammation score,intestinal barrier function and intestinal epithelial cell pyroptosis.The mechanism of nodakenin protection against pyroptosis of intestinal epithelial cells was explored using network pharmacology analysis and in vivo and in vitro experiments.Results In LPS-and ATP-induced colonic organoids,treatment with nodakenin significantly inhibited the expressions of NLRP3,GSDMD-N,cleaved caspase-1 and caspase-11,improved intestinal FITC-dextran(FD4,4000)permeability,and decreased the levels of IL-1β and IL-18.In the mouse model of TNBS-induced colitis,nodakenin treatment significantly alleviated weight loss,reduced DAI score,inflammatory cell infiltration and inflammation score,and decreased serum FD4 and I-FABP levels and bacteria translocation to the mesenteric lymph nodes,spleen and liver.The mice with nodakenin treatment had also lowered expressions of NLRP3,GSDMD-N,cleaved caspase-1 and caspase-11 in the intestinal mucosa.Network pharmacology analysis suggested that the inhibitory effect of nodakenin on colitis was associated with the PI3K/Akt pathway.In both the colonic organoid model and mouse models of colitis,nodakenin effectively inhibited the activation of the PI3K/Akt pathway,and the application of IGF-1,a PI3K/Akt pathway activator,strongly attenuated the protective effect of nodakenin against intestinal epithelial cell pyroptosis and intestinal barrier dysfunction.Co

关 键 词：克罗恩病 紫花前胡苷 细胞焦亡 肠上皮细胞 PI3K/AKT 

分 类 号：R285.5[医药卫生—中药学]