摘要
目的:研究参莲(SL)提取物在脂质过氧化炎症损伤模型中,对巨噬细胞功能的作用和促进炎症消散的药效和药理机制。方法:利用氧化低密度脂蛋白(ox-LDL)体外诱导小鼠巨噬细胞RAW264. 7细胞和人单核细胞THP-1造成脂质过氧化损伤模型,酶联免疫吸附测定(ELISA)检测不同质量浓度的SL提取物(2. 5,5. 0,10. 0,20. 0 mg·L^(-1))对巨噬细胞肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β),单核细胞趋化蛋白-1(MCP-1)的含量;流式细胞术检测巨噬细胞泡沫化形成;transwell实验检测SL提取物对THP-1趋化功能的影响;以及用蛋白免疫印迹法(Western blot)检测诱导型一氧化氮合酶(i NOS),炎症消散因子5脂氧合酶(ALOX5)以及对核转录因子-κB(NF-κB)信号通路中的磷酸化p65(p-p65)和磷酸化IκK(p-IκK)蛋白表达情况。结果:与正常组比较,ox-LDL提高M1型巨噬细胞标志物TNF-α,IL-1β,i NOS的表达(P <0. 01),抑制M2型标志物IL-10的表达,促进泡沫细胞形成(P <0. 01),诱导THP-1向ox-LDL趋化,提高MCP-1分泌量(P <0. 01),抑制ALOX5的表达。与模型组比较,SL提取物可以降低TNF-α,IL-1β,i NOS的表达(P <0. 01),提高IL-10的表达(P <0. 05);能显著降低巨噬细胞吞噬脂质后泡沫化的形成(P <0. 01);能降低THP-1细胞MCP-1的分泌(P <0. 01),减少趋化的细胞数目(P <0. 01);促进炎症消散因子ALOX5的升高(P <0. 05),同时抑制p65和IκK磷酸化表达(P <0. 01)。结论:在脂质过氧化炎症损伤模型中,SL提取物具有诱导巨噬细胞M2极化,抑制巨噬细胞对ox-LDL的趋化和泡沫化的形成,提高ALOX5表达,促进炎症消散,从而改善脂质过氧化炎症损伤状态,上述功能与抑制NF-κB信号通路中p65和IκK磷酸化水平有关。
Objective:To study the effect of Shenlian extract(SL extract)on macrophage function and inflammatory resolution in lipid peroxidation inflammatory injury models.Method:The effects of different concentrations of SL extract(2.5,5.0,10.0,20.0 mg·L-1)on the polarization type,foam formation and chemotactic function of macrophages were detected with RAW264.7 cells induced by oxidized low-density lipoprotein(ox-LDL).Western blot was used to detect pro-inflammatory resolution factor arachidonate5-lipoxygenase(ALOX5)and inducible inducible nitric oxide synthase(i NOS),and phosphorylated p65(p-p65)and phosphorylated IκK(p-IκK)in nuclear factor(NF)-κB related signaling pathways.Result:Compared with the control group,ox-LDL enhanced the expressions of M1 macrophage markers TNF-α,IL-1β,i NOS(P<0.01),inhibited the expressions of IL-10 of M2 markers,promoted foam cell formation(P<0.01),induced THP-1 chemotaxis ability,increased the content of MCP-1(P<0.01),and inhibited the expression of ALOX5.Compared with the model group,SL extract reduced the expressions of TNF-α,IL-1β,and i NOS(P<0.01),increased the expression of IL-10(P<0.05),and down-regulated the formation of foam of macrophages(P<0.01).In transwell assay,SL extract obviously decreased the MCP-1 secretion and the number of chemotaxis cells(P<0.01),and promoted the increase of the inflammatory resolution factor ALOX5.The phosphorylation of p65 and IκK was inhibited significantly(P<0.01).Conclusion:In the inflammatory damage model of lipid peroxidation,SL extract can regulate the polarization of macrophage,inhibit the chemotaxis and foaming of ox-LDL,increase the inflammatory resolution molecular expression,and improve the state of lipid peroxidation,which may be related to the inhibition of NF-κB signaling pathway.
作者
尹婕
李琦
赵正
杨庆
刘思思
李玉洁
陈颖
王娅杰
翁小刚
蔡维艳
朱晓新
YIN Jie;LI Qi;ZHAO Zheng;YANG Qing;LIU Si-si;LI Yu-jie;CHEN Ying;WANG Ya-jie;WENG Xiao-gang;CAI Wei-yan;ZHU Xiao-xin(School of Traditional Chinese Medicine,Capital Medical University,Beijing 100069,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第10期26-32,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81573649)
国家"重大新药创制"科技重大专项(2017ZX09101002-002-002
2017ZX09101002-002-008)
中国中医科学院国际合作项目(GH2017-01
ZXKT17010)
