基于NF-κB信号通路研究鸡娃草石油醚提取物对LPS诱导RAW264.7细胞的抗炎作用  

Stuy on anti-inflammatory effect of petroleum ether extract of Plumbagella micrantha on LPS-induced RAW264.7 cells through NF-κB signaling pathway

作　　者：莫婷 何婷 杨丽娟 杨森 康菊英 马静 赵芳芳 火明才 卢海瑞 MO Ting;HE Ting;YANG Lijuan;YANG Sen;KANG Juying;MA Jing;ZHAO Fangfang;HUO Mingcai;LU Hairui(Ningxia research institute of Hui medicine,Yinchuan Ningxia 750021)

机构地区：[1]宁夏回族医药研究所,宁夏银川750021

出　　处：《广西中医药》2024年第2期67-73,共7页Guangxi Journal of Traditional Chinese Medicine

基　　金：宁夏自然科学基金项目(编号:2023AAC03718);宁夏回族自治区卫生健康系统科研课题(编号:2021-NW-059);宁夏中药资源保障能力提升项目(宁卫办发﹝2023﹞34号)。

摘　　要：目的:观察鸡娃草石油醚提取物(Pm-PE)通过核因子κB(NF-κB)信号通路对脂多糖(LPS)诱导RAW264.7细胞的抗炎作用。方法:以不同浓度Pm-PE作用于RAW264.7细胞24 h,分析在LPS刺激下药物对细胞形态变化的影响;采用CCK-8法检测不同浓度的Pm-PE对RAW264.7细胞活性的影响;以一氧化氮(NO)试剂盒法检测致RAW264.7炎症细胞的NO释放量;以酶联免疫吸附法(ELISA)检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和白介素-10(IL-10)的分泌;以逆转录聚合酶链反应法(RT-PCR)检测促炎因子诱导型一氧化氮合酶(iNOS)、TNF-α、IL-6和抗炎因子IL-10的mRNA表达情况;以免疫印迹法(Western blot)检测细胞NF-κB p65、IκB-α蛋白的表达。结果:与LPS模型组比较,Pm-PE不同剂量组能剂量依赖性显著降低NO、TNF-α、IL-6的分泌水平和下调iNOS、TNF-α、IL-6 mRNA的表达,同时调节抗炎因子IL-10的表达;Pm-PE中、高剂量组能显著性抑制NF-κB p-p65和p-IκB-α蛋白的相对表达水平。结论:鸡娃草石油醚提取物可通过NF-κB信号通路减少LPS诱导的RAW264.7细胞炎症相关因子的释放,从而发挥抗炎作用。Objective:To observe the anti-inflammatory effect of petroleum ether extract of Plumbagella micrantha(Pm-PE)on lipopolysaccharide(LPS)-induced RAW264.7 cells through nuclear factor-κB(NF-κB)signaling path⁃way.Methods:RAW264.7 cells were treated with different concentrations of Pm-PE for 24 h,and the effects of drugs on cell morphology under LPS stimulation were analyzed.The effect of different concentrations of Pm-PE on the activi⁃ty of RAW264.7 cells was detected by Cell Counting Kit-8(CCK-8).The NO release of RAW264.7 inflammatory cells was detected by nitric oxide(NO)kit method.The secretion of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and interleukin-10(IL-10)was detected by enzyme-linked immunosorbent assay(ELISA).Reverse transcription poly⁃merase chain reaction(RT-PCR)was used to detect pro-inflammatory factors.The mRNA expression of inducible ni⁃tric oxide synthase(iNOS),TNF-α,IL-6 and anti-inflammatory factor IL-10 were detected by RT-PCR.The expres⁃sion of NF-κB p65 and IκB-αprotein was detected by Western blot.Results:Compared with the LPS model group,Pm-PE at different doses could significantly reduce the dose dependence.NO,TNF-α,IL-6 secretion and iNOS,TNF-α,IL-6 mRNA expression,while regulating the expression of anti-inflammatory factor IL-10;the medium and high dose groups of Pm-PE could significantly inhibit the relative expression levels of NF-κBp-p65 and p-IκB-αprotein.Conclusion:The petroleum ether extract of Plumbagella micrantha can reduce the release of inflammation-related fac⁃tors in RAW264.7 cells induced by LPS through NF-κB signaling pathway,thus exerting anti-inflammatory effects.

关 键 词：鸡娃草 石油醚提取物 小鼠巨噬细胞系 脂多糖 抗炎 NF-ΚB信号通路 

分 类 号：R285.5[医药卫生—中药学]