摘要
目的探讨表没食子儿茶素没食子酸酯(EGCG)对SSA/Ro52(Ro52)介导的人单核-巨噬细胞株THP-1凋亡及炎症因子分泌的影响。方法构建稳定Ro52过表达质粒(p CMV-Ro52),p CMVRo52转染THP-1细胞48 h,检测Ro52 m RNA和蛋白表达水平的变化,采用膜联蛋白-荧光素异硫氰酸酯和碘化丙锭双染检测THP-1细胞凋亡率,酶联免疫吸附法检测肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、IL-13水平。p CMV-Ro52质粒转染48 h后,分别使用0、5、10、20和50 mg/L EGCG处理细胞24 h,检测Ro52表达的变化、THP-1细胞的凋亡,以及TNF-α、IL-1β、IL-13水平。结果 p CMVRo52促进Ro52表达(P<0.05),上调THP-1细胞凋亡率及TNF-α、IL-1β、IL-13水平(P<0.05);在Ro52过表达的THP-1细胞中,与0 mg/L EGCG组相比,10、20和50 mg/L EGCG组Ro52蛋白表达水平、凋亡率及TNF-α、IL-1β、IL-13水平降低(P<0.05)。结论 Ro52过表达可促进人单核-巨噬细胞THP-1凋亡及炎症因子TNF-α、IL-1β、IL-13分泌;EGCG可减弱Ro52介导的THP-1细胞凋亡,减少炎症因子TNF-α、IL-1β、IL-13的分泌。
Objective To explore the influence of epigallocatechin gallate(EGCG)on apoptosis of human monocyte macrophage cell line(THP-1)and expression of inflammatory cytokines through Ro52 overexpression.Methods Ro52 over-expression plasmid(PCMV-Ro52,20μg)was constructed and transfected into THP-1 cells for 48 h.Real-time fluorescent quantitative PCR was used to detect the mRNA level of Ro52 and the protein expression level of Ro52 was detected by Western blot.The apoptosis of THP-1 cells was detected by Annexin V-FITC and propidium iodide(PI)staining.The levels of TNF-α,IL-1βand IL-13 were measured by ELISA.After 48 h of pCMV-Ro52 plasmid transfection,the cells were treated with EGCG at different concentrations(0,5,10,20 and 50 mg/L)for 24 h.The protein expression level of Ro52,the expression levels of TNF-α,IL-1βand IL-13 and the apoptosis rate of THP-1 cells were detected.Results pCMV-Ro52(20μg)significantly increased the mRNA and protein levels of Ro52(P<0.05)and up-regulated the apoptosis rate of THP-1 cells and the levels of TNF-α,IL-1βand IL-13(P<0.05).In the Ro52-overexpressed THP-1 cells,compared with the 0 mg/L group,the Ro52 protein levels,the cell apoptosis rates and the levels of inflammatory cytokines(TNF-α,IL-1βand IL-13)were significantly decreased in the 10 mg/L,20 mg/L and 50 mg/L EGCG-treated groups(P<0.05).Conclusions The over-expression of Ro52 could promote the apoptosis of THP-1 cells and the secretion of TNF-α,IL-1βand IL-13.EGCG can attenuate Ro52-induced apoptosis of THP-1 cells and secretion of inflammatory cytokines.
作者
张懿
刘畅
Yi Zhang;Chang Liu(Jinzhou Medical University,Jinzhou,Liaoning 121000,China;Department of Endocrinology,the First Affiliated Hospital of Jinzhou Medical University,Jinzhou,Liaoning 121000,China)
出处
《中国现代医学杂志》
CAS
2018年第26期38-44,共7页
China Journal of Modern Medicine
