葛根素抑制巨噬细胞NLRP3炎症小体表达减轻溃疡性结肠炎及其初步机制分析  

Puerarin inhibits expression of macrophage NLRP3 inflammasome to alleviate ulcerative colitis and preliminary analysis of its mechanism

作　　者：赵鑫 陈旭涛 鲁星妤 但文利 高国俊 唐康 陈丽华 ZHAO Xin;CHEN Xutao;LU Xingyu;DAN Wenli;GAO Guojun;TANG Kang;CHEN Lihua(College of Medical Technology,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Department of Immunology,Basic Medical Science Academy,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]陕西中医药大学医学技术学院,咸阳712046 [2]空军军医大学基础医学院免疫学教研室,西安710032

出　　处：《中国免疫学杂志》2025年第4期775-782,共8页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(82071848)。

摘　　要：目的:探讨葛根素(PUE)对巨噬细胞NLRP3炎症小体表达的影响及其对小鼠溃疡性结肠炎(UC)的作用。方法:建立葡聚糖硫酸钠(DSS)诱导的小鼠UC模型,连续7 d通过灌胃给予PUE后测定小鼠的体质量、疾病活动指数、结肠长度;HE染色观察组织病理损伤;流式细胞术检测小鼠外周血单个核细胞及结肠固有层巨噬细胞比例;免疫荧光检测NLRP3炎症小体与巨噬细胞的共定位情况;qRT-PCR检测小鼠结肠组织促炎细胞因子和NLRP3炎症小体相关基因的mRNA表达水平;Western blot检测结肠组织ZO-1、Occludin、cleaved caspase-1及IL-1β蛋白表达水平;利用脂多糖(LPS)及三磷酸腺苷(ATP)构建细胞模型,qRT-PCR检测NLRP3炎症小体相关基因的mRNA表达水平;Western blot检测PUE对NLRP3炎症小体相关蛋白表达水平及NF-κB信号通路的影响。结果:PUE干预显著改善DSS诱导的小鼠UC症状,包括体质量、疾病指数、结肠长度和病理损伤;PUE干预UC小鼠后,小鼠结肠固有层Ly6C+MHCⅡ-单核来源的巨噬细胞浸润减少;结肠组织中促炎细胞因子及NLRP3炎症小体相关分子表达水平降低;小鼠肠上皮细胞间ZO-1及Occludin表达水平升高;体外细胞实验证实,PUE能够降低LPS结合ATP诱导的巨噬细胞NLRP3炎症小体相关分子表达水平,同时降低p-NF-κB p65蛋白表达水平。结论:PUE可以通过减轻肠道组织炎症、修复上皮屏障显著改善UC症状。其机制可能是通过调控NF-κB信号通路,减少了巨噬细胞内NLRP3炎症小体的形成与激活。Objective:To investigate the effect of natural plant compound puerarin(PUE)on expression of NLRP3 inflammasome in macrophages and its effect in ulcerative colitis(UC)in mice.Methods:A mouse model of UC was established using dextran sulfate sodium(DSS).Mice received PUE via gavage for 7 consecutive days,body weight,disease activity index and colon length were measured.HE staining was performed to assess tissue pathological damage.Flow cytometry was used to determine the proportion of peripheral blood mononuclear cells in colonic lamina propria.Immunofluorescence was employed to assess the colocalization of NLRP3 inflammasome and macrophages.qRT-PCR was conducted to measure mRNA expression levels of pro-inflammatory cytokines and NLRP3-related genes in colonic tissues.Protein expression levels of ZO-1,Occludin,cleaved caspase-1 and IL-1βin colonic tissues were detected by Western blot.A cell model was established using lipopolysaccharide(LPS)and adenosine triphosphate(ATP).mRNA expression levels of genes related to NLRP3 inflammasome were detected by qRT-PCR.Western blot was used to detect effects of PUE on expression levels of proteins related to NLRP3 inflammasome and NF-κB signaling pathway.Results:PUE treatment significantly improved symptoms of DSS-induced UC in mice,including body weight,disease index,colon length and pathological damage.Following PUE intervention,infiltration of Ly6C+MHCⅡ−monocyte derived macrophages in the colonic lamina propria was reduced.Expression levels of pro-inflammatory cytokines and NLRP3 inflammasome related molecules in colonic tissues were decreased.PUE treatment increased expression levels of ZO-1 and Occludin in intestinal epithelial cells.In vitro experiments confirmed that PUE reduced expression levels of NLRP3 inflammasome-related molecules in macrophages induced by LPS combined with ATP,as well as protein expression level of p-NF-κB p65.Conclusion:PUE significantly alleviates the symptoms of UC by reducing intestinal tissue inflammation and repairing the epithelial

关 键 词：溃疡性结肠炎 葛根素 巨噬细胞 NLRP3炎症小体 

分 类 号：R285.5[医药卫生—中药学]