绿原酸调控巨噬细胞极化治疗溃疡性结肠炎的机制研究  

Mechanism Research of Chlorogenic Acid on Treating Ulcerative Colitis via Regulating Macrophage Polarization

作　　者：李跃文 万强[3] LI Yuewen;WAN Qiang(Jinhua Traditional Chinese Medicine Hospital,Jinhua,321017,Zhejiang China;National Famous Traditional Chinese Medicine Expert SHI Weiqun Inheritance Studio;Affiliated Hospital of Jiangxi University of Chinese Medicine)

机构地区：[1]金华市中医医院,浙江金华321017 [2]全国名老中医药专家施维群传承工作室 [3]江西中医药大学附属医院

出　　处：《浙江中医药大学学报》2023年第9期969-975,1001,共8页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金项目(81660770);江西省卫生健康委科技计划项目(202310664);江西省教育厅科学技术研究项目(GJJ190654);金华市科技计划项目(2020-4-057)。

摘　　要：[目的]探讨绿原酸(chlorogenic acid,CGA)对葡聚糖硫酸钠(dextran sulfate sodium,DSS)诱导的溃疡性结肠炎(ulcerative colitis,UC)的治疗作用及分子机制。[方法]C57BL/6小鼠50只,用3%DSS腹腔注射7 d,建立UC动物模型并随机分为5组,除UC模型组外,其余4组分别给予柳氮磺胺吡啶(sulfasalazine,SZ)100 mg/kg·d和CGA 30、60、120 mg/kg·d灌胃10 d,另以C57BL/6小鼠10只设为正常对照组。检测小鼠疾病活动指数(disease activity index,DAI);以苏木精-伊红(hematoxylin-eosin,HE)染色观察结肠组织病理变化;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测结肠组织中肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)、IL-10、IL-1β水平;定量实时聚合酶链式反应(quantitative real timepolymerase chain reaction,qRT-PCR)检测结肠组织中诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)、精氨酸酶-1(arginase-1,Arg-1)的mRNA表达;免疫印迹检测结肠组织小鼠无翅型乳腺肿瘤病毒整合位点家族成员5a(wingless type 5a,Wnt5a)、C1型尼曼-匹克蛋白(Nieman-Pick type C1,NPC1)的蛋白表达;流式细胞术检测结肠组织M1型及M2型巨噬细胞比例。[结果]与正常对照组比较,UC模型组DAI评分显著增高,结肠组织出现明显炎症病理改变,M1型巨噬细胞比例,TNF-α、IL-6、IL-1β水平,iNOS mRNA表达,Wnt5a及NPC1蛋白表达均显著增高,M2型巨噬细胞比例、IL-10水平及Arg-1 mRNA表达均显著降低(P<0.01)。与UC模型组比较,CGA可显著降低小鼠DAI评分,减轻结肠组织炎症病理改变,降低结肠组织M1型巨噬细胞比例,TNF-α、IL-6、IL-1β水平,iNOS mRNA表达,以及Wnt5a和NPC1蛋白表达,增加M2型巨噬细胞比例、IL-10水平及Arg-1 mRNA表达(P<0.05,P<0.01)。[结论]CGA可通过减少巨噬细胞向M1型极化,从而减轻由DSS诱导的炎症反应以治疗UC,其机制与抑制Wnt5a/NPC1信号通路活化有关。[Objective]To investigate the therapeutic effect and molecular mechanism of chlorogenic acid(CGA)on dextan sodium sulfate(DSS)-induced ulcerative colitis(UC).[Methods]Fifty C57BL/6 mice were intraperitoneally injected with 3%DSS for 7 days to establish the animal model of UC and were randomly divided into 5 groups.Except UC model group,the other 4 groups were separately given sulfasalazine(SZ)100 mg/kg·d and CGA 30,60,120 mg/kg·d by gavage for 10 days,and 10 C57BL/6 mice were set as normal control group.Disease activity index(DAI)was detected;hematoxylin-eosin(HE)staining was used to observe the pathological changes of colon tissues;the levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-10(IL-10)and interleukin-1β(IL-1β)in colon tissues were detected by enzyme linked immunosorbent assay(ELISA);the mRNA expressions of inducible nitric oxide synthase(iNOS)and arginase-1(Arg-1)in colon tissues were detected by quantitative real time-polymerase chain reaction(qRT-PCR);the expressions of wingless type mouse mammary tumor virus integration site family type 5a(Wnt5a)and Nieman-Pick type C1(NPC1)in colon tissues were detected by Western blot;the proportion of M1 and M2 macrophages in colon tissues was determined by flow cytometry.[Results]Compared with normal control group,the DAI score of UC model group was significantly increased,and there were obvious inflammatory pathological changes in colon tissue,proportion of M1 macrophages,TNF-α,IL-6,IL-1βlevels,iNOS mRNA expression,Wnt5a and NPC1 protein expression were significantly increased,while proportion of M2 macrophages,IL-10 level,and Arg-1 mRNA expression were significantly decreased(P0.01).Compared with UC model group,CGA significantly decreased DAI score,attenuated inflammatory pathological changes in colon tissue,decreased proportion of M1 macrophages,TNF-α,IL-6,IL-1βlevels,iNOS mRNA expression,Wnt5a and NPC1 protein expression,and increased proportion of M2 macrophages,IL-10 level and Arg-1 mRNA expression were observed in colon tis

关 键 词：绿原酸 葡聚糖硫酸钠 溃疡性结肠炎 Wnt5a/NPC1信号通路 巨噬细胞极化 炎症反应 

分 类 号：R331[医药卫生—人体生理学]