苏荠宁黄酮通过抑制PI3K/AKT信号通路拮抗肠上皮细胞凋亡改善小鼠实验性结肠炎  

Moslosooflavone ameliorates dextran sulfate sodium-induced colitis in mice by suppressing intestinal epithelium apoptosis via inhibiting the PI3K/AKT signaling pathway

作　　者：储菲 陈孝华 宋博文 杨晶晶 左芦根 CHU Fei;CHEN Xiaohua;SONG Bowen;YANG Jingjing;ZUO Lugen(Department of Pharmacy,First Affiliated Hospital of Bengbu Medical University,Bengbu 233004,China;Department of Gastrointestinal Surgery,First Affiliated Hospital of Bengbu Medical University,Bengbu 233004,China;Anhui Key Laboratory of Basic and Translational Research on Inflammation-related Diseases,Bengbu 233030,China)

机构地区：[1]蚌埠医科大学第一附属医院药剂科,安徽蚌埠233004 [2]蚌埠医科大学第一附属医院胃肠外科,安徽蚌埠233004 [3]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233030

出　　处：《南方医科大学学报》2025年第4期819-828,共10页Journal of Southern Medical University

基　　金：国家自然科学基金(82370534);安徽省卫生健康科研项目(AHWJ2022a019);安徽省高校杰出青年科研项目(2022AH020085);安徽省生化药物工程技术研究中心开放课题(2022SYKFD04);安徽省高校中青年教师培养行动项目(JWFX2023032);蚌埠医科大学研究生科研创新项目(Byycxz24020)。

摘　　要：目的探讨苏荠宁黄酮(MOS)改善实验性结肠炎的作用和机制。方法通过C57BL/6J小鼠饮用2.5%葡聚糖硫酸钠(DSS)诱导实验性结肠炎,并采用腹腔注射MOS(200 mg/kg)进行干预。实验小鼠共分为4组(n=6):WT、WT+MOS、DSS和DSS+MOS组,通过检测小鼠体质量、结肠长度、HE染色、肠屏障功能和TUNEL染色来评估MOS对拮抗结肠炎和肠上皮细胞凋亡的作用。体外通过脂多糖(LPS,100μg/mL)刺激小鼠结肠类器官,并采用MOS(120μmol/L)进行干预。体外实验共分为4组:Control、Control+MOS、LPS和LPS+MOS组,以评估MOS对LPS诱导的肠屏障损伤和炎症反应的药理作用。网络药理学分析MOS作用的功能学途径和分子机制,并联合免疫印迹检测验证细胞凋亡相关蛋白的表达及其调控机制。结果体内实验表明MOS治疗改善DSS小鼠的体质量减轻(P<0.05)、DAI评分(P<0.05)、结肠缩短(P<0.05)、结肠组织炎症评分(P<0.05)和促炎因子(TNF-α、IL-1β、IL-6和IFN-γ,P<0.05)的表达。肠屏障功能检测表明,MOS治疗降低血中的FITC-Dextran(P<0.05)和I-FABP的浓度,并增加了肠上皮细胞间紧密连接蛋白的表达(ZO-1和claudin-1,P<0.05)。体外实验显示MOS治疗能够改善LPS诱导的结肠类器官的炎症因子水平和肠上皮细胞屏障的损伤(P<0.05)。免疫印迹结果表明,MOS干预在体内和体外都下调C-caspase3和BAX的表达(P<0.05),并上调抗凋亡蛋白Bcl-2的表达(P<0.05)。机制分析表明,DSS小鼠结肠组织和LPS刺激的结肠类器官的PI3K和AKT蛋白的磷酸化水平都被MOS抑制(P<0.05)。结论MOS可能通过PI3K/AKT信号抑制肠上皮细胞凋亡,从而改善肠道屏障的完整性并缓解实验性结肠炎。Objective To investigate the effect of moslosooflavone(MOS)for ameliorating dextran sulfate sodium(DSS)-induced colitis in mice and the underlying molecular mechanism.Methods C57BL/6J mice with or without DSS exposure in the drinking water were both randomized into two groups for treatment with intraperitoneal injections with MOS(200 mg/kg)or normal saline for 7 days(n=6).Disease severity of the mice was assessed by observing changes in body weight,colon length,histopathology(HE staining),intestinal barrier function,and TUNEL staining.In the in vitro studies,lipopolysaccharide(LPS)-stimulated mouse colon organoids were treated with MOS(120μmol/L)for 24 h,and the changes in barrier dysfunction and inflammation were analyzed.Network pharmacology and Western blotting were employed to identify functional pathways and apoptotic protein regulation associated with the therapeutic effect of MOS on colitis.Results In the mouse models of DSSindcued colitis,MOS treatment significantly reduced body weight loss,disease activity index(DAI)scores and colon shortening,ameliorated colonic histopathological changes and inflammation,and lowered pro-inflammatory cytokine levels(TNF-α,IL-1β,IL-6,and IFN-γ).MOS effectively restored intestinal barrier integrity in the mice by reducing serum FITCdextran and I-FABP concentrations while enhancing the tight junction proteins(ZO-1 and claudin-1).In the colon organoids,MOS significantly suppressed LPS-induced inflammatory responses and epithelial barrier disruption.Western blotting revealed that MOS downregulated C-caspase-3 and BAX and upregulated Bcl-2 expressions in both models.Mechanistically,MOS suppressed PI3K and AKT phosphorylation in both DSS-treated mouse colonic tissues and LPS-stimulated organoids.Conclusion MOS alleviates experimental colitis in mice by inhibiting intestinal epithelial apoptosis via inhibiting the PI3K/AKT pathway,thereby restoring intestinal barrier integrity and reducing inflammation.

关 键 词：炎症性肠病 苏荠宁黄酮 肠上皮细胞凋亡 PI3K/AKT信号 

分 类 号：R285.5[医药卫生—中药学]