健脾益肠散对TNF-α诱导的树突状细胞NLRP3炎症小体信号通路表达的影响  

Effect of Jianpi Yichang Powder on the Expression of NLRP3 Inflammasome Signaling Pathway of Dendritic Cells Induced by TNF-α

作　　者：蔺晓源[1] 陈龙彪 罗沛文 周赛男[1] 刘杰民[2] LIN Xiaoyuan;CHEN Longbiao;LUO Peiwen;ZHOU Sainan;LIU Jiemin(The First 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：《中国中医药信息杂志》2024年第10期109-114,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金地区科学基金(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)诱导的树突状细胞(DCs)NLRP3炎症小体信号通路相关蛋白表达的影响,探讨其治疗溃疡性结肠炎的作用机制。方法分离培养C57BL/6小鼠原代DCs,分为正常组、模型组、阴性对照组、阳性药组和中药组,TNF-α诱导构建DCs炎症微环境模型,各组分别予相应处理后,采用CCK-8法检测细胞增殖能力,免疫荧光、Western blot分别检测DCs核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)、胱天蛋白酶-1(Caspase-1)蛋白表达,Western blot、RT-PCR分别检测DCs白细胞介素(IL)-1β、IL-18蛋白和mRNA表达。结果与正常组比较,模型组DCs增殖能力显著下降(P<0.01),细胞NLRP3、ASC、Caspase-1阳性表达显著升高,NLRP3、ASC、Caspase-1、IL-18、IL-1β蛋白表达及IL-18、IL-1βmRNA表达显著升高(P<0.01);与模型组和阴性对照组比较,阳性药组和中药组DCs增殖能力显著上升(P<0.01),细胞NLRP3、ASC、Caspase-1阳性表达显著降低(P<0.01),NLRP3、ASC、Caspase-1、IL-1β、IL-18蛋白表达显著降低(P<0.05,P<0.01),IL-1β、IL-18mRNA表达显著降低(P<0.01),且中药组细胞增殖能力、Caspase-1阳性表达、IL-18 mRNA表达均明显高于阳性药组(P<0.05),IL-1βmRNA表达明显低于阳性药组(P<0.05)。结论健脾益肠散能抑制TNF-α诱导的DCs炎性反应,其治疗溃疡性结肠炎作用机制可能与调控NLRP3炎症小体信号通路NLRP3、ASC、Caspase-1、IL-18、IL-1β表达有关。Objective To investigate the effects of Jianpi Yichang Powder on regulating the NLRP3 inflammasome signaling pathway to inhibit dendritic cells(DCs)inflammatory injury induced by TNF-α;To explore its mechanism in the treatment of ulcerative colitis.Methods C57BL/6 mice DCs were primitively isolated and cultured.The cultured DCs were randomly divided into the normal group,model group,negative control group,positive drug group and TCM group.DCs inflammatory microenvironment model was established by TNF-αinduction.After corresponding treatments were given to each group,CCK-8 method was used to detect cell proliferation ability,immunofluorescence and Western blot were used to detect the protein expressions of NLRP3,ASC and Caspase-1 in DCs respectively,Western blot and RT-PCR were used to detect the protein and mRNA expressions of IL-1βand IL-18 in DCs.Results Compared with the normal group,the proliferation ability in DCs of model group significantly decreased(P<0.01),the positive expressions of NLRP3,ASC and Caspase-1 significantly increased,the expressions of NLRP3,ASC,Caspase-1,IL-18,IL-1βprotein and IL-18,IL-1βmRNA in DCs significantly increased(P<0.01).Compared with the model group and negative control group,the proliferation ability of DCs significantly increased in positive drug group and TCM group(P<0.01),the positive expressions of NLRP3,ASC,and Caspase-1 significantly decreased(P<0.01),the protein expressions of NLRP3,ASC,Caspase-1,IL-1βand IL-18 significantly decreased(P<0.05,P<0.01),and the mRNA expressions of IL-1βand IL-18 significantly decreased(P<0.01).The cell proliferation ability,Caspase-1 positive expression,and IL-18 mRNA expression of TCM group were significantly higher than those of the positive drug group(P<0.05),while the mRNA expression of IL-1βwas lower than that of the positive drug group(P<0.05).Conclusion Jianpi Yichang Powder can inhibit the inflammatory response of DCs induced by TNF-α,and its mechanism in the treatment of ulcerative colitis may be related to regulating the e

关 键 词：健脾益肠散 溃疡性结肠炎 树突状细胞 NLRP3炎症小体 

分 类 号：R285.5[医药卫生—中药学]