野菊花水提物对RAW264.7炎症细胞模型的抗炎作用及其机制  

Anti-inflammatory Effect and Mechanismof Chrysanthemum Indicum Decoction on RAW264.7 Inflammatory Cell Model

作　　者：熊鑫[1] 黄传奇[1] 程璐[1] XIONG Xin;HUANG Chuanqi;CHENG Lu(Department of Pharmacy,Wuhan No.1 Hospital,Wuhan 430022,China)

机构地区：[1]武汉市第一医院药学部,武汉430022

出　　处：《医药导报》2024年第8期1192-1198,共7页Herald of Medicine

基　　金：2020年武汉市卫生健康委面上项目一般项目(WZ20D06);2022年武汉药学会-定向科研(WHPA2022020032)。

摘　　要：目的建立以脂多糖(LPS)诱导的RAW 264.7巨噬细胞为模型,探讨野菊花提取液(CID)通过核转录因子(NF-κB)信号通路发挥抗炎活性的作用及其分子机制。方法以噻唑蓝(MTT)法检测不同浓度CID对RAW 264.7巨噬细胞活性的影响以筛选适宜的实验浓度;分别采用Griess法和酶联免疫吸附试验(ELISA)测定50、100、200μg·mL^(-1) CID干预后各组细胞中一氧化氮(NO)和肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的释放量;实时荧光定量聚合酶链式反应(RT-PCR)分析各组中环氧合酶-2(COX-2)和诱导型一氧化氮合酶(iNOS)mRNA的相对表达水平;免疫印迹实验(WB)观察各组中nuclear factor-kappa B p65(NF-κB p65)、inhibitor kappa B(IκB-α)和磷酸化IκB-α(p-IκB-α)的蛋白表达。结果50~200μg·mL^(-1)的CID可显著降低LPS诱导RAW264.7巨噬细胞中NO、TNF-α和IL-6的生成量(P<0.01),并能下调COX-2和iNOX mRNA的相对表达(P<0.01)、下调p-IκB-α、总的NF-κB p65、细胞核NF-κB p65的蛋白相对含量(P<0.01),并上调IκB-α、细胞质NF-κB p65的相对含量(P<0.01)。结论CID可有效降低LPS诱导RAW 264.7巨噬细胞的炎症因子释放,其机制可能与通过减少TNF-α等关键蛋白表达以及通过抑制NF-κB等炎症信号通路激活来抑制炎症发生有关。Objective To investigate the anti-inflammatory effect and molecular mechanism of Chrysanthemum indicum decoction(C.indicum decoction,CID)through NF-κB signaling pathway based on RAW264.7 macrophages model induced by lipopolysaccharide(LPS).Methods MTT was used to select the appropriate concentration of CID on the activity of RAW 264.7 macrophages;Griess method and enzyme-linked immunosorbent assay(ELISA)were used to measure the release of nitric oxide(NO),tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in cells treated with 50,100 and 200μg·mL^(-1) CID,respectively;and the relative contents expression levels of cyclooxygenase-2(COX-2)and inducible nitric oxide synthase(iNOS)in each group were analyzed by real-time fluorescent quantitative polymerase chain reaction(Real-time PCR);the relative production of nuclear factor-kappa B p65(NF-κB p65),inhibitor kappa B(IκB-α)and phosphorylated IκB-α(p-IκB-α)in each group was observed by Western blotting(WB).Results 50-200μg·mL^(-1) CID significantly decreased the relative production of NO,TNF-α,and IL-6 in RAW264.7 macrophages induced by LPS(P<0.01),down-regulated the expression of COX-2 and iNOX mRNA(P<0.01),down-regulated the relative contents of p-IκB-α,total NF-κB p65 and nuclear NF-κB p65(P<0.01),and up-regulated the relative contents of IκB-αand cytoplasmic NF-κB p65(P<0.01).Conclusion CID could effectively inhibit the release of inflammatory factors from RAW 264.7 macrophages induced by LPS,and the mechanism may be related to the inhibition of inflammation by reducing the expression of TNF-αand other key proteins and regulating the inflammatory signaling pathway such as NF-κB.

关 键 词：野菊花 抗炎作用 RAW264.7炎症细胞模型 脂多糖 核转录因子-ΚB 

分 类 号：R978.1[医药卫生—药品]