金线莲提取物对LPS诱导RAW264.7细胞炎性因子和MAPK信号通路的影响  

Study on Anti-inflammatory Mechanism in Vitro of Jinxianlian(Anoectochilus roxburghii)Extract

作　　者：朱亮 王群星[1] 徐菲拉[1] ZHU Liang;WANG Qunxing;XU Feila(Jinhua Municipal Central Hospital,Jinhua,Zhejiang 321000)

机构地区：[1]金华市中心医院,浙江金华321000

出　　处：《中国中医药科技》2025年第3期415-420,共6页Chinese Journal of Traditional Medical Science and Technology

基　　金：浙江省金华市科技计划项目公益类项目(2021-4-281);浙江省医学会项目(2020ZYC-A126)。

摘　　要：目的:探讨金线莲提取物对脂多糖诱导RAW264.7细胞炎症反应的抗炎作用及其作用机制。方法:采用MTT法分析金线莲对RAW264.7细胞活力的影响。通过脂多糖处理RAW264.7细胞24 h建立细胞炎症模型,实验组造模前1 h用金线莲200、100、50 mg/L预处理,ELISA测定RAW 264.7细胞上清液白细胞介素6(IL-6)、单核细胞趋化蛋白-1(MCP-1)、活性白细胞介素12(IL-12/p70)、干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)和白细胞介素10(IL-10)水平;免疫荧光检测各组RAW 264.7细胞磷酸化乙酰辅酶A羧化酶(p-ACC)、磷酸化细胞外信号调节激酶(p-ERK)和磷酸化核转录因子κB亚基(p-p65)的表达;Western blot检测RAW 264.7细胞p-ERK、磷酸化c-Jun氨基末端激酶(p-JNK)、磷酸化丝裂原活化蛋白激酶p38(p-p38)、磷酸化腺苷酸-活化蛋白激酶α(p-AMPKα)、髓过氧化物酶(MPO)蛋白表达水平。结果:与空白组比较,不同剂量金线莲提取物对RAW 264.7细胞活性无影响。LPS刺激后,模型组细胞中细胞上清液IL-6、MCP-1、IL-12p70、IFN-γ、TNF-α和IL-10含量明显增加,细胞内p-ACC、p-ERK、p-p65、p-JNK、p-p38、p-AMPKα及MPO蛋白表达显著升高;中高剂量金线莲提取物能显著降低细胞上清液IL-6、MCP-1、IL-12p70、IFN-γ、TNF-α、IL-10含量;显著降低RAW 264.7细胞p-ERK、p-JNK、p-p38、p-AMPKα、MPO蛋白表达水平;中剂量组对p-ACC的蛋白表达量均无显著性影响。结论:金线莲提取物能可有效抑制脂多糖所致炎症反应,其作用机制可能与调节MAPK信号通路减轻炎症反应有关。Objective:To investigate the anti-inflammatory effect and mechanism of Jinxianlian(Anoectochilus roxburghii)on LPS-induced inflammatory response in RAW264.7 cells.Methods:The effect of Jinxianlian on the viability of RAW264.7 cells was analyzed by MTT method.RAW264.7 cells were cultured with LPS for 24 h to establish a cellular inflammation model,the cells were given different doses(200,100,50 mg/L)of JXL solution pre-cultured 1 h before modeling.The levels of IL-6,MCP-1,IL-12p70,IFN-γ,TNF-αand IL-10 in the supernatant of RAW 264.7 cells were measured by ELISA,The expressions of p-ACC,p-ERK and p-p65 in RAW 264.7 cells were detected by immunofluorescence.The expression levels of p-ERK,p-JNK,p-p38,p-AMPKαand MPO in RAW 264.7 cells were detected by Western blot.Results:Compared with the the blank control group,the cell activity of RAW 264.7 was not affected by different doses of JXL solution.After LPS stimulation,the concentrations of IL-6,MCP-1,IL-12p70,IFN-γ,TNF-αand IL-10 in the cell supernatant of the model group were increased significantly,and the expressions of p-ACC,p-ERK,p-p65,p-JNK,p-p38,p-AMPKαand MPO proteins were increased significantly.Medium and high doses of JXL solution could significantly reduce the concentrations of IL-6,MCP-1,IL-12p70,IFN-γ,TNF-αand IL-10 in cell supernatants and the expression levels of p-ERK,p-p65,p-JNK,p-p38,p-AMPKαand MPO proteins in cell supernatants,but there was no significant change in the expression of p-ACC proteins in the medium dose group.Conclusion:JXL solution can effectively inhibit the inflammatory response caused by LPS,and its mechanism of action may be related to the regulation of MAPK signaling pathway to reduce inflammatory response.

关 键 词：金线莲 RAW 264.7细胞 炎性因子 MAPK信号通路 体外实验 

分 类 号：R285.5[医药卫生—中药学]