白芍总苷对TNF-α诱导Caco-2炎性细胞模型的作用机制  

Effect of Total Glucosides of Paeony on TNF-αMechanism of Inducing Caco-2 Inflammatory Cell Model

作　　者：朱占荣 官杰[1] 吴艳敏[1] 王琪[1] 罗晓庆[1] 张贺 王慧[1] ZHU Zhanrong;GUAN Jie;WU Yanmin(Department of Immunology,Qiqihar Medical University,Heilongjiang 161006,China)

机构地区：[1]齐齐哈尔医学院医学技术学院,161006

出　　处：《医学研究杂志》2022年第12期64-68,共5页Journal of Medical Research

基　　金：黑龙江省教育厅科学技术研究项目(2017-KYYWF-0733)。

摘　　要：目的研究白芍总苷(total glucosides of paeony,TGP)对肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)诱导的人结肠上皮Caco-2炎性细胞自噬和凋亡水平的影响,并探究其保护机制。方法将对数生长期的Caco-2细胞分为对照组、模型组和给药组。其中对照组细胞常规培养,模型组和给药组细胞TNF-α100ng/ml刺激48h建立炎性细胞模型;给药组细胞使用不同浓度TGP培养,对照组和模型组细胞使用等量完全培养基培养;用噻唑蓝(MTT)法检测细胞增殖情况;利用酶联免疫吸附试验检测各组中白介素1β(interleukin 1β,IL-1β)和白介素6(interleukin 6,IL-6)蛋白质水平表达变化;采用Western blot法检测程序性死亡受体1(Beclin-1)、微管相关蛋白轻链3B(LC3B)、Bcl-2蛋白的表达。结果TGP对Caco-2炎性细胞模型的增殖具有抑制作用,与正常组比较,TNF-α诱导的Caco-2炎性细胞中IL-1β、IL-6的表达明显增高(P<0.05);与模型组比较,TGP能够下调IL-1β、IL-6的表达水平,Beclin-1、LC3B的表达水平降低,Bcl-2表达水平升高,差异有统计学意义(P<0.05)。结论TGP能够下调IL-1β、IL-6的表达水平,抑制Beclin-1、LC3B蛋白表达水平,上调抗凋亡基因Bcl-2表达,TGP对TNF-α诱导Caco-2炎性细胞模型的治疗作用与抑制炎症及调节肠上皮细胞自噬有关。Objective To investigate the effect of total glucosides of paeony(TGP)on the autophagy and apoptosis levels of TNF-αinduced human colonic epithelial Caco-2 inflammatory cells,and to explore its protective mechanism.Methods The Caco-2 cells in logarithmic growth phase were divided into control group,model group and administration group.Among them,the cells in the control group were routinely cultured,and the cells in the model group and the treatment group were cultured with TNF-α100ng/ml to establish an inflammatory cell model;the cells in the treatment group were cultured with total glucosides of different concentrations of paeony,and the cells in the control group and the model group were cultured with the same amount of complete culture.The cell proliferation was detected by thiazolyl blue(MTT)method;the protein levels of interleukin 1β(IL-1β)and interleukin 6(IL-6)in each group were detected by enzyme-linked immunosorbent assay;Western blot was used to detect the expression of Beclin-1,LC3B and Bcl-2 proteins.Results The expression level of autophagy in Caco-2 inflammatory cells was increased.Compared with the model group,total glucosides of paeony could down-regulate the expression levels of IL-1βand IL-6,inhibit the protein expression levels of Beclin-1 and LC3B,and up-regulate the anti-apoptotic expression levels.The expression of apoptotic gene Bcl-2 was significantly different(P<0.05).Conclusion TGP can down regulate IL-1β,The expression level of IL-6 inhibits the expression level of beclin-1 and LC3B protein and up regulates the expression of anti apoptotic gene Bcl-2.This regulation depends on autophagy of intestinal epithelial cells.

关 键 词：白芍总苷 炎症性肠病 肿瘤坏死因子α 细胞自噬 

分 类 号：R285.5[医药卫生—中药学]