健脾益肠散对TNF-α处理树突状细胞炎症因子水平及共刺激分子CD40表达的影响  

Effect of Jianpi Yichang Powder on the Levels of Inflammatory Factor and theExpression of Co-Stimulatory Molecule CD40 in Dendritic Cells Treated by TNF-α

作　　者：陈龙彪 蔺晓源[1] 刘杰民[2] 周赛男[1] CHEN Longbiao;LIN Xiaoyuan;LIU Jiemin;ZHOU Sainan(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Guizhou Provincial People's Hospital,Guiyang 550002,China)

机构地区：[1]湖南中医药大学第一附属医院,湖南长沙410007 [2]贵州省人民医院,贵州贵阳550002

出　　处：《中医药学报》2024年第9期15-21,共7页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金项目(82060850);贵州省高层次创新型人才项目(黔科合平台人才〔2020〕6016-2)。

摘　　要：目的:观察健脾益肠散对肿瘤坏死因子α(TNF-α)处理的树突状细胞(DC)炎症因子水平及其刺激分子CD40表达的影响。方法:原代分离、培养C57BL/6小鼠DC细胞,采用流式细胞术对DC的主要标记物CD11c进行鉴定,并将培养的DCs随机分为正常组、模型组、阴性对照组、阳性药组及受试药组,TNF-α处理细胞构建DC炎症微环境模型,各组给予不同处理后,于倒置荧光显微镜下观察各组DCs的形态,采用CCK-8检测细胞的增殖能力,采用ELISA法检测各组DC培养上清液中CD40、IL-1β及IL-18含量,采用Western blot及RT-PCR技术分别检测各组DCs中CD40的蛋白和mRNA表达水平。结果:分离纯化的DC的CD11c表达率为85.1%,与正常组比较,模型组DC培养上清液中CD40、IL-1β及IL-18的含量、细胞中CD40的蛋白及其mRNA表达均明显增多,而增殖能力显著下降(P<0.01);受试药组DC培养上清液中CD40、IL-1β及IL-18的含量、CD40的蛋白及其mRNA表达减少,且细胞增殖能力上升,与模型组及阴性对照组相比较皆有统计学意义(P<0.01)。结论:健脾益肠散对TNF-α处理的DCs共刺激分子CD40的蛋白和mRNA表达及炎症因子的含量具有明显的抑制作用,可能是其发挥免疫耐受作用治疗溃疡性结肠炎的重要机制之一。Objective:To investigate the effect of Jianpi Yichang powder(JYP)on the expression of co-stimulatory molecule CD40 and the levels of inflammatory factor in dendritic cell(DC)treated by TNF-αin vitro.Methods:Primary C57BL/6 mouse DCs were isolated and cultured.The main marker of DC,CD11c,was identified by flow cytometry,and the cultured DCs were randomly divided into normal group,model group,negative control group,positive drug group and test drug group.Cells were treated by TNF-αto construct the inflammatory microenvironment model of DC.After different treatments,the morphology of DCs in each group was observed under inverted fluorescence microscope,and the proliferation ability of DCs was detected by CCK-8.The proliferation capacity of DCs was detected by CCK-8,the contents of CD40,IL-1βand IL-18 in DC culture supernatant of each group were detected by ELISA,and the expression levels of CD40 protein and mRNA in DCS of each group were detected by Western blot and RT-PCR,respectively.Results:The CD11c expression rate of purified DC was 85.1%.Compared with normal group,the contents of CD40,IL-1βand IL-18 in DC culture supernatant,the expression of CD40 protein and its mRNA in cells in model group significantly increased,while the proliferation ability significantly decreased(P<0.01).The contents of CD40,IL-1βand IL-18,and the expression of CD40 protein and its mRNA in DC culture supernatant of test group decreased,and the cell proliferation ability increased,which had statistical significance compared with model group and negative control group(P<0.01).Conclusion:Jianpi Yichang powder can significantly inhibit the expression of TNF-αtreated and Co-Stimulatory Molecule CD40 protein and mRNA and the content of inflammatory factors.It may be one of the important mechanisms by which it exerts immune tolerance in the treatment of ulcerative colitis.

关 键 词：溃疡性结肠炎 健脾益肠散 树突状细胞 CD40 炎症因子 

分 类 号：R285.5[医药卫生—中药学]