川藏香茶菜庚素的体外抗炎作用与分子机制研究  

Anti-inflammatory effect and mechanism of Pseurata H on inflammation model of LPS-induced RAW264.7 cells

作　　者：杨东娟[1] 查广才[1] 邹湘辉[1] 吴丹彤 伍清婷 YANG Dongjuan;ZHA Guangcai;ZOU Xianghui;WU Dantong;WU Qingting(School of Life Sciences and Food Engineering,Hanshan Normal University,Chaozhou 521041,China)

机构地区：[1]韩山师范学院生命科学与食品工程学院,潮州521041

出　　处：《生物学杂志》2024年第2期39-43,共5页Journal of Biology

基　　金：韩山师范学院一般项目(LY202601);广东省粤东药食资源功能物质与治未病研究重点实验室(2021B1212040015);广东省普通高校重点科研项目(2022ZDZX4030)。

摘　　要：为探究川藏香茶菜庚素(Pseurata H)对巨噬细胞炎症模型的抗炎作用及分子机制,利用LPS刺激RAW264.7细胞建立体外炎症模型,并分别以浓度为10、20和40μmol/L的Pseurata H干预该细胞模型。利用CCK-8法检测Pseurata H对细胞增殖活性的影响;Griess法和ELISA试剂盒分别检测细胞上清液中NO、IL-6和TNF-α的含量;Western Blot检测COX-2、iNOS蛋白、p-NF-κB和p-ERK蛋白的表达变化;以RT-qPCR法检测药物干预前后细胞中iNOS、TNF-α、IL-6和IL-1β的mRNA基因表达变化。结果表明:浓度为10、20和40μmol/L的Pseurata H对细胞没有毒性,不同浓度的Pseurata H对LPS诱导RAW264.7细胞炎症模型中NO、IL-6和TNF-α分泌具有很好的下调作用,对iNOS、COX-2、NF-κB、p-NF-κB、ERK和p-ERK蛋白的表达具有良好的抑制作用,且具有浓度效应。Pseurata H具有明显的抗炎作用,其作用机制可能是通过抑制NF-κB/MAPK信号通路而发挥抗炎作用。To study the anti-inflammatory effect and mechanism of Pseurata H on peritoneal macrophages(RAW264.7 cells)inflammation models,RAW264.7 cells were stimulated with LPS to establish an inflammation model in vitro,and Pseurata H with the concentrations of 10,20,and 40μmol/L was used to interfere with this cell model.The cytotoxicity of Pseurata H in RAW264.7 cells was determined by CCK-8 colorimetric assay.Griess assay was employed to detect the content of NO in the supernatant.The release of IL-6 and TNF-αin the supernatant was assayed using ELISA.The protein expressions of COX-2,iNOS,p-NF-κB and the p-ERK were measured by Western Blot.The mRNA expressions of iNOS,TNF-α,IL-6 and IL-1βwere analyzed by RT-qPCR.The results showed that Pseurata H(10,20,40μmol/L)displayed no apparent cytotoxicity in RAW264.7 cells.The release levels of NO,iNOS,TNF-α,and IL-6 were decreased by Pseurata H(10,20,40μmol/L).In addition,Pseurata H also inhibited the expressions of iNOS,COX-2,NF-κB,p-NF-κB,ERK,and p-ERK proteins with concentration effect.Pseurata H has a significant anti-inflammatory effect,and its mechanism of action may be through the inhibition of NF-κB/MAPK signaling pathway to exert anti-inflammatory effects.

关 键 词：川藏香茶菜庚素 RAW264.7细胞 脂多糖 炎症 NF-κB/MAPK通路 

分 类 号：Q753[生物学—分子生物学] R285[医药卫生—中药学]