基于TLR4/NF-κB/NLRP3信号通路的马齿苋生物碱对溃疡性结肠炎作用机制  

Mechanism of Purslane Alkaloids on Ulcerative Colitis Based on TLR4/NF-κB/NLRP3 Signalling Pathway

作　　者：宋英英 郑佳慧 艾金霞 杨志平[3] SONG Yingying;ZHENG Jiahui;AI Jinxia;YANG Zhiping(School of Clinical Medicine,Beihua University,Jilin 132013,China;School of Medical Technology,Beihua University,Jilin 132013,China;Affiliated Hospital of Beihua University,Jilin 132011,China)

机构地区：[1]北华大学临床医学院,吉林吉林132013 [2]北华大学医学技术学院,吉林吉林132013 [3]北华大学附属医院,吉林吉林132011

出　　处：《北华大学学报(自然科学版)》2025年第3期322-328,共7页Journal of Beihua University(Natural Science)

基　　金：吉林省科技发展计划项目(YDZJ202301ZYTS507);北华大学研究生创新计划项目(2023080);北华大学医学技术学院“迈瑞”学生创新项目(20210010)。

摘　　要：目的探讨马齿苋生物碱(purslane alkaloids,PA)对小鼠溃疡性结肠炎(ulcerative colitis,UC)的治疗作用及调节机制。方法将35只小鼠随机分为空白对照组、模型组、药物治疗组(马齿苋生物碱低、中、高剂量组,美沙拉嗪组,马齿苋生物碱联合美沙拉嗪组)。前7 d空白对照组小鼠给予纯净水,模型组及药物治疗组给予3%硫酸葡聚糖钠盐(dextran sulfate sodium salt,DSS)溶液,构建UC动物模型。从第8天起,空白对照组及模型组给予生理盐水,药物治疗组小鼠开始接受固定剂量药物灌胃处理,持续14 d。在此期间,每日对小鼠健康状况、体质量、粪便特征及是否有血便进行详细记录。试验结束后计算各组小鼠疾病活动指数(disease activity index,DAI),测量结肠长度;采用苏木素-伊红染色观察结肠组织病理变化;酶联免疫吸附试验(ELISA)法检测血清及结肠组织中促炎因子白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量;蛋白印迹试验(Western blot)、实时荧光定量PCR(RT-qPCR)测定结肠组织Toll样受体4(Toll-like receptor 4,TLR4)、核因子kappa-B(nuclear factor kappa-B,NF-κB)及Nod样受体蛋白3(Nod-like receptor3,NLRP3)蛋白及mRNA表达。结果与模型组比较,马齿苋生物碱治疗组及联合组能缓解小鼠症状,增加体质量(P<0.05,P<0.01);降低DAI评分(P<0.05,P<0.01);缓解结肠组织缩短(P<0.01)及病理损伤;降低小鼠血清和结肠组织中IL-1β、IL-6、TNF-α炎症水平(P<0.05,P<0.01),显著下调结肠组织中TLR4、NF-κB和NLRP3蛋白及mRNA表达水平(P<0.05,P<0.01)。结论马齿苋生物碱对小鼠急性UC有明显治疗作用,能降低UC小鼠结肠组织促炎细胞因子水平,明显缓解病理损伤,其抗炎机制可能与抑制TLR4/NF-κB/NLRP3信号通路有关。Objective To investigate therapeutic effects and regulatory mechanisms of purslane alkaloids(PA)on ulcerative colitis(UC)in mice.Methods Thirty-five mice were randomly divided into control group,model group and drug treatment group(which included purslane alkaloids groups at low,medium and high doses,mesalazine group,as well as purslane alkaloids combined with mesalazine group).The UC animal model was constructed by providing pure water to the mice in control group and 3%dextran sulfate sodium salt(DSS)solution to the mice in model and drug treatment groups for the first 7 days.From day 8 onward,saline was administered to control and model groups,while the mice in drug treatment group began receiving fixed-dose drug gavage treatment for 14 days.During this period,the health status,body mass,fecal characteristics and the presence or absence of bloody stools were recorded daily.At the end of experiment,the disease activity index(DAI)was calculated for each group of mice,and the length of colon was measured.Hematoxylin-eosin staining was used to observe histopathological changes in the colon.Enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of pro-inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6)and tumour necrosis factor-α(TNF-α)in serum and colon tissues.The protein and mRNA exprossion of Toll-like receptor 4(TLR4),nuclear factor kappa-B(NF-κB)and Nod-like receptor3(NLRP3)of colon tissues were measured by Western blot and real-time fluorescence quantitative PCR(RT-qPCR).Results Compared with model group,purslane alkaloids treatment group and combined group could alleviate symptoms and gain body mass in mice(P<0.05,P<0.01),reduce DAI score(P<0.05,P<0.01),alleviate shortening of colon tissues(P<0.01)and pathological damage,and reduce the levels of inflammation of IL-1β,IL-6 and TNF-αin the serum and colon tissues of mice(P<0.05,P<0.01).Additionally,there was a significant down-regulation of the protein and mRNA expression levels of TLR4,NF-κB and NLRP3 in colon tissues(P<0.0

关 键 词：马齿苋生物碱 溃疡性结肠炎 Toll样受体4 核因子kappa-B Nod样受体蛋白3 

分 类 号：R285.5[医药卫生—中药学] R574.6[医药卫生—中医学]