表没食子儿茶素没食子酸酯抑制MAPK/NF-κB通路减轻脂多糖诱导的RAW 264.7细胞炎症反应  

Epigallocatechin gallate inhibits MAPK/NF-κB pathway and reduces lipopolysaccharide-induced inflammatory response in RAW 264.7 cells

作　　者：邹俊 杜逸亭 Zou Jun;Du Yiting(Department of Emergency,Women’s and Children’s Hospital Affiliated to Medical College of University of Electronic Science and Technology of China,Chengdu Women’s and Children’s Central Hospital,Chengdu Sichuan 610073,China)

机构地区：[1]电子科技大学医学院附属妇女儿童医院·成都市妇女儿童中心医院急诊科,四川成都610073

出　　处：《遵义医科大学学报》2022年第3期310-316,共7页Journal of Zunyi Medical University

基　　金：成都市卫生局科学研究基金资助项目(NO:20140776)。

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)调控丝裂原活化蛋白激酶/核因子κB(MAPK/NF-κB)信号通路对脂多糖(LPS)诱导的RAW 264.7细胞炎症反应的影响。方法取对数生长期的RAW264.7细胞,随机分为对照组、LPS(1μg/mL)组、LPS+EGCG-L(12.5μmol/L)组、LPS+EGCG-M(25μmol/L)组、LPS+EGCG-H(50μmol/L)组、LPS+DEX(阳性对照,0.5μg/mL)组、LPS+EGCG-H+Anisomycin(300 ng/mL MAPK/NF-κB通路激活剂Anisomycin)组,细胞在添加对应的药物处理1 h后,再加入100μL 1μg/mL LPS,培养24h后,进行后续相关实验。利用倒置显微镜观察各组细胞的形态特征;MTT法检测各组细胞的存活率;Griess法检测各组细胞上清中一氧化氮(NO)含量;ELISA法检测各组细胞上清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)的表达;Western blotting检测诱导型一氧化氮合酶(iNOS)、环氧化酶-2(COX-2)蛋白及MAPK/NF-κB通路相关蛋白的表达。结果各组细胞的存活率均大于92%;与对照组比较,LPS组RAW264.7细胞形态呈梭形,细胞中iNOS、COX-2、p-p38、p-IκBα、p-NF-κB蛋白及上清中NO含量、TNF-α、IL-6、IL-1β表达显著升高,IL-10表达显著降低(P<0.05);与LPS组比较,LPS+EGCG-L组、LPS+EGCG-M组、LPS+EGCG-H组、LPS+DEX组对应指标变化与上述相反,且EGCG浓度越高,趋势越明显(P<0.05);Anisomycin可逆转高剂量EGCG对LPS诱导RAW264.7细胞炎症反应的抑制作用。结论EGCG可抑制LPS诱导的RAW 264.7细胞炎症反应,该机制与抑制MAPK/NF-κB信号通路有关。Objective To explore the effect of epigallocatechin gallate(EGCG)regulating mitogen-activated protein kinase/nuclear factor-κB(MAPK/NF-κB)signaling pathway on the inflammatory response of RAW 264.7 cells induced by lipopolysaccharide(LPS).Methods RAW264.7 cells in logarithmic growth phase were randomly divided into control group,LPS(1μg/ml)group,LPS+EGCG-L(12.5μmol/L)group,LPS+EGCG-M(25μmol/L)group,LPS+EGCG-H(50μmol/L)group,LPS+DEX(positive control,0.5μg/ml)group,and LPS+EGCG-H+Anisomycin(300 ng/ml MAPK/NF-κB pathway activator Anisomycin)group.Cells were treated with the corresponding drugs for 1 hour,and then 100μl of 1μg/ml LPS was added.After 24 hours of culture,the follow-up related experiments were carried out:An inverted microscope was used to observe the morphological characteristics of each group of cells;MTT method to detect the survival rate of each group of cells;Griess method to detect the content of nitric oxide(NO)in the supernatant;ELISA method to detect the tumor necrosis factor-α(TNF-α),Interleukin-6(IL-6),Interleukin-1β(IL-1β),Interleukin-10(IL-10)expression in the supernatant;Western blot was used to detect inducible monoxide Nitrogen synthase(iNOS),cyclooxygenase-2(COX-2)protein and MAPK/NF-κB pathway related protein expression.Results The survival rate of the cells in each group was greater than 92%;compared with the control group,the RAW264.7 cells in the LPS group showed a spindle shape,the expression iNOS,COX-2,p-p38,p-IκBα,and p-NF-κB proteins in cells,and the expression of NO,TNF-α,IL-6 and IL-1βin the cell supernatant was significantly increased,and the expression of IL-10 was decreased(P<0.05);Compared with the LPS group,the corresponding index changes in the LPS+EGCG-L group,LPS+EGCG-M group,LPS+EGCG-H group,and LPS+DEX group were alleviated,and the higher the EGCG concentration,the more obvious the trend(P<0.05);Anisomycin can reverse the inhibitory effect of high-dose EGCG on LPS-induced inflammatory response in RAW264.7 cells.Conclusion EGCG can inhibit the infl

关 键 词：炎症 表没食子儿茶素没食子酸酯 丝裂原活化蛋白激酶/核因子κB通路 脂多糖 RAW264.7细胞 

分 类 号：R725[医药卫生—儿科]