康复新液抑制NF-κB通路及巨噬细胞M1型极化减轻IBD小鼠结肠炎的作用机制OA
Mechanism of colitis alleviation by inhibiting the nuclear factor-κB pathway and macrophage M1 polarization via Kangfuxin liquid in inflammatory bowel disease mice
目的 基于炎症性肠病(IBD)的病理机制与核因子κB(NF-κB)信号通路的过度激活及巨噬细胞M1 型极化密切相关,利用比较研究探索IBD疾病过程中康复新液对NF-κB通路及巨噬细胞M1 型极化的调节机制.方法 通过硫酸葡聚糖钠(DSS)诱导构建小鼠 IBD 模型,将 48 只 C57BL/6 小鼠分为对照(Control)、模型(Model)、柳氮磺吡啶(SASP)、康复新液低剂量(5 mL/kg,KFX-L)、康复新液中剂量(10 mL/kg,KFX-M)、康复新液高剂量(20 mL/kg,KFX-H)、NF-κB抑制剂预处理(NF-κBi-pretreated)及NF-κB抑制剂预处理+康复新液高剂量(NF-κBi-pretreated+KFX-H)组,共 8 组.记录其疾病活动指数(DAI)、结肠黏膜损伤指数(CMDI)、组织病理学评分,同时还利用RT-qPCR法检测结肠组织CD86、肿瘤坏死因子-α(TNF-α)、磷酸化IκBα(p-IκBα)和IL-1β的mRNA表达水平,蛋白免疫印迹(Western blot)法检测结肠组织CD86、iNOS、p-IκBα的蛋白表达水平,流式细胞术分析康复新液对疾病模型的影响.结果 DSS诱导成功构建了小鼠IBD模型,各项研究指标的测得值均在Model组最高(P<0.05).药物干预后,除SASP 组CD86 下调程度较小并与Model组比较差异无统计学意义(P>0.05)外,各组的各项指标与 Model 组比较均显著降低(P<0.05),而康复新液干预的 3 组中以KFX-H组的指标下调最为明显(P<0.05),但NF-κB抑制剂与高剂量康复新液联合干预后并未明显增加疗效,各指标测得值与药物单独干预各组间的差异无统计学意义(P>0.05).结论 康复新液、柳氮磺吡啶和NF-κB抑制剂对DSS诱导的IBD模型小鼠结肠炎症均具有保护作用,但其作用机制存在差异.康复新液与NF-κB抑制剂可通过降低IκBα磷酸化水平抑制NF-κB通路活化,并下调巨噬细胞M1 型极化,从而协同减轻肠道炎症反应;而柳氮磺吡啶对巨噬细胞M1 极化的影响较弱.
Objective The pathological mechanism of inflammatory bowel disease(IBD)is closely related to overactivation of the nuclear factor-κB(NF-κB)signaling pathway and M1 polarization of macrophages.Accordingly,this study aimed to investigate the effects of Kangfuxin solution on the regulatory mechanism of the NF-κB pathway during the progression of IBD.Methods A mouse model of IBD was established using dextran sodium sulfate(DSS).Forty-eight C57BL/6 mice were divided into eight groups:Control,Model,SASP,KFX-L(5 mL/kg),KFX-M(10 mL/kg),KFX-H(20 mL/kg),NF-κBi-pretreated and NF-κBi-pretreated+KFX-H groups.The disease activity index(DAI),colonic mucosal damage index(CMDI),and histopathological scores were recorded.mRNA expression levels of CD86,tumor necrosis factor-α(TNF-α),phosphorylated NF-κB inhibitor α(p-IκBα),and interleukin(IL)-1β in colonic tissues were detected by RT-qPCR,and protein expression levels of CD86,inducible nitric oxide synthase,and p-IκBα in colon tissues were detected by Western blot.Flow cytometry was used to analyze the effect of Kangfuxin liquid on the disease model.Results DSS-induction successfully established a mouse IBD model,with the measured values of various research indicators being highest in the Model group(P<0.05).The indicators in each group,except the SASP group,were significantly decreased compared with the Model group(P<0.05),while downregulation of CD86 in the SASP group was not significant(P>0.05).Among the three Kangfuxin liquid groups,the indicators were significantly reduced in the KFX-H group(P<0.05).The combined intervention of NF-κB inhibitor and high-dose Kangfuxin liquid did not significantly enhance the therapeutic effect,with no significant differences in the measured values of each indicator compared with the single-drug groups(P>0.05).Conclusions Kangfuxin liquid,sulfasalazine and NF-κB inhibitors all have protective effects on colonic inflammation in DSS-induced IBD model mice,but their sites of action differ.Kangfuxin liquid and NF-κB inhibitors synergistically attenuate the intestinal inflammatory response by inhibiting NF-κB pathway activation through reducing IκBα phosphorylation,as well as down-regulating macrophage M1-type polarization.In contrast,sulfasalazine exerts a weaker effect on macrophage M1 polarization.
陈立;李捷;张杉杉;王宁;吴学东
大理大学第一附属医院小儿外科,云南 大理 671000大理大学第一附属医院小儿外科,云南 大理 671000大理大学第一附属医院小儿外科,云南 大理 671000大理大学第一附属医院小儿外科,云南 大理 671000大理大学第一附属医院小儿外科,云南 大理 671000
医药卫生
炎症性肠病NF-κB通路巨噬细胞极化调控机制康复新液
inflammatory bowel diseaseNF-κB signaling pathwaymacrophage polarizationregulatory mechanismKangfuxin liquid
《中国比较医学杂志》 2026 (1)
22-33,12
云南省教育厅科学研究基金项目(2024Y894).
评论