基于NF-κB/Akt信号通路探讨犁头草醇提物对LPS诱导RAW264.7细胞的抗炎作用  

Anti-inflammatory effects of Viola inconspicua ethanol extract on LPS-induced RAW264.7 cells based on NF-κB/Akt signal pathway

作　　者：秦慧真 林思 张淼[1] 谢凤凤 罗静 朱华[1] QIN Hui-zhen;LIN Si;ZHANG Miao;XIE Feng-feng;LUO Jing;ZHU Hua(Guangxi Key Laboratory of Zhuang and Yao Ethnic Medicine,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]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》2022年第12期3811-3817,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);广西民族药资源与应用工程研究中心(桂发改高技函[2020]2605号);广西产5种壮瑶药材质量标准与产品开发研究(桂科计字[2021]196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);广西中医药大学-中南大学壮瑶药研究联合实验室(桂科计字[2021]238号)。

摘　　要：目的探究犁头草醇提物对脂多糖(LPS)诱导的小鼠单核巨噬细胞RAW264.7体外抗炎作用及其相关机制。方法采用MTT法检测不同质量浓度犁头草醇提物对RAW264.7细胞活性的影响,以筛选实验用剂量。以LPS诱导RAW264.7细胞进行造模,给予40、80、120μg/mL犁头草醇提物进行干预,Griess法检测细胞上清NO水平,ELISA法检测细胞上清IL-1β、IL-6、TNF-α水平,RT-qPCR法检测细胞IL-1β、IL-6、TNF-αmRNA表达,Western blot法检测细胞iNOS、COX-2、p65、p-p65、IκBα、p-IκBα、Akt、p-Akt蛋白表达。结果犁头草醇提物在20~120μg/mL质量浓度范围内对RAW264.7细胞无抑制作用。与模型组比较,犁头草醇提物各剂量组能降低NO水平,IL-1β、IL-6、TNF-α水平和mRNA表达,以及iNOS、COX-2、p-p65、p-IκBα、p-Akt蛋白表达(P<0.05,P<0.01)。结论犁头草醇提物能够降低LPS诱导的RAW264.7细胞炎症,其分子机制与抑制NF-κB/Akt信号通路有关。AIM To explore the anti-inflammatory effects and mechanism of Viola inconspicua ethanol extract on mouse mononuclear macrophage RAW264.7 cells induced by lipopolysaccharide(LPS)in vitro.METHODS The V.inconspicua ethanol extract at different concentrations had its effect on the viability of RAW264.7 cells detected by MTT assay for the screening of the experimental dosage.The LPS-induced RAW264.7 cell models were intervened with 40,80 and 120μg/mL V.inconspicua ethanol extract,respectively.Subsequently,they had their NO level in cell supernatant detected by Griess method;their IL-1β,IL-6 and TNF-αlevels in cell supernatant detected by ELISA;their mRNA expressions of IL-1β,IL-6 and TNF-αdetected by RT-qPCR;and their protein expressions of iNOS,COX-2,p65,p-p65,IκBα,p-IκBα,Akt and p-Akt detected by Western blot.RESULTS No inhibitory effect of V.inconspicua ethanol extract was observed on RAW264.7 cells within the concentration range of 20-120μg/mL.Compared with the model group,each group intervened with V.inconspicua ethanol extract shared reduced NO level,levels and mRNA expressions of IL-1β,IL-6,TNF-α,and the protein expressions of iNOS,COX-2,p-p65,p-IκBαand p-Akt(P<0.05,P<0.01).CONCLUSION The V.inconspicua ethanol extract can reduce the inflammation of LPS-induced RAW264.7 cells,and its molecular mechanism is related to the inhibition of NF-κB/Akt signal pathway.

关 键 词：犁头草醇提物 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]