乌蔹莓醇提物对LPS诱导RAW264.7细胞炎症模型的抗炎作用  

Anti-inflammatory effects of ethanolic extract of Cayratia japonica on LPS-induced inflammation model of RAW264.7 cells

作　　者：龙莉 秦慧真 林思 朱华 韦安达 LONG Li;QIN Hui-zhen;LIN Si;ZHU Hua;WEI An-da(Centre for Preparations of Zhuang and Yao Ethnic Medicines,Guangxi International Zhuang Medicine Hospital,Nanning 530201,China;Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西国际壮医医院壮瑶药制剂中心,广西南宁530201 [2]广西中医药大学壮瑶药重点实验室,广西南宁530200

出　　处：《中成药》2023年第8期2525-2531,共7页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(82060695);广西壮瑶药重点实验室项目(桂科基字[2014]32号);广西壮瑶药协同创新中心项目(桂教科研[2013]20号);广西第八批特聘专家项目——壮瑶药质量标准研究(桂人才通字〔2019〕13号);广西科技基地和人才专项(桂科AD21238031);广西重点研发计划项目(桂科AB21196016);广西壮族自治区民族药资源与应用工程研究中心(桂发改高技函[2020]2605号)。

摘　　要：目的探讨乌蔹莓70%醇提物对脂多糖(LPS)诱导RAW264.7细胞炎症模型的作用及其机制。方法MTT法观察不同质量浓度乌蔹莓醇提物对RAW264.7细胞活性的影响,筛选合适的实验剂量。以LPS(1μg/mL)诱导建立细胞炎症模型,同时加入30、60、120μg/mL乌蔹莓醇提物进行处理,24 h后Griess法检测细胞上清液NO水平,DCFH-DA荧光探针检测细胞内ROS水平,ELISA法检测细胞上清液PGE_(2)、TNF-α、IL-6、IL-1β水平,RT-qPCR法检测细胞TNF-α、IL-6、IL-1βmRNA表达,Western blot法检测细胞iNOS、COX-2、NF-κB p65、p-NF-κB p65、Akt、p-Akt、p38 MAPK、p-p38 MAPK蛋白表达。结果乌蔹莓醇提取物质量浓度在180μg/mL及以下时对RAW264.7细胞存活率无明显变化(P>0.05)。与模型组比较,乌蔹莓醇提物组细胞内ROS水平及上清液NO、PGE_(2)、TNF-α、IL-6、IL-1β水平均降低(P<0.01),细胞TNF-α、IL-6、IL-1βmRNA表达降低(P<0.05,P<0.01),iNOS、COX-2蛋白表达均下调(P<0.05,P<0.01),p-NF-κB p65/NF-κB p65、p-Akt/Akt比值均降低(P<0.05,P<0.01),但p-p38 MAPK/p38 MAPK比值无明显变化(P>0.05)。结论乌蔹莓醇提物可拮抗LPS诱导的小鼠巨噬细胞炎症反应,其机制可能与抑制NF-κB/Akt信号通路,进而抑制炎症相关因子的表达有关。AIM To explore the effects and mechanism of 70%ethanolic extract of Cayratia japonica on lipopolysaccharide(LPS)-induced inflammation model of RAW264.7 cells.METHODS MTT method was used to observe the effects of ethanolic extract of C.japonica at different concentrations on the activity of RAW264.7 cells,and to screen the appropriate experimental dose.RAW264.7 cells were induced into the inflammation models by LPS(1μg/mL)and simultaneusly underwent 30,60 and 120μg/mL of ethanolic extract of C.japonica treatment respectively.24 hours later,the cell models had their NO level in cell supernatant detected by Griess method;their intracellular ROS level detected by DCFH-DA fluorescent probes;and their levels of PGE_(2),TNF-α,IL-6 and IL-1βin cell supernatant detected by ELISA;their mRNA expressions of TNF-α,IL-6 and IL-1βdetected by RT-qPCR;and their protein expressions of iNOS,COX-2,NF-κB p65,p-NF-κB p65,Akt,p-Akt,p38 MAPK and p-p38 MAPK detected by Western blot.RESULTS Ethanolic extract of C.japonica under concentration of 180μg/mL brought no significant change in the survival rate of RAW264.7 cells(P>0.05).Compared with the model group,the ethanolic extract groups displayed decreased ROS level in cells and levels of NO,PGE_(2),TNF-α,IL-6 and IL-1βin supernatant(P<0.01);decreased mRNA expressions of TNF-α,IL-6 and IL-1βin cells(P<0.05,P<0.01);and decreased ratios of p-NF-κB p65/NF-κB p65 and p-Akt/Akt(P<0.05,P<0.01),but no significant change of the ratio of p-p38 MAPK/p38 MAPK(P>0.05).CONCLUSION The ethanolic extract of C.japonica can antagonize LPS-induced inflammatory reaction of mouse macrophages,and its mechanism may lie in the inhibition of the expressions of inflammation-related factors via NF-κB/Akt signaling pathway.

关 键 词：乌蔹莓 醇提物 脂多糖 RAW264.7细胞 炎症 NF-κB/Akt信号通路 

分 类 号：R285.5[医药卫生—中药学]