土荆皮乙酸对脂多糖诱导的RAW264.7巨噬细胞炎症反应及M1表型偏移的抑制作用  

Inhibitory effects of pseudolaric acid B on inflammatory response and M1 phenotype polarization in RAW264.7 macrophages induced by lipopolysaccharide

作　　者：李玉秀[1,2] 李覃[2,3] 姬文婕[2] 李宵[2] 马永强[2] 赵季红[2] 周欣[2] 李玉明[2] 

机构地区：[1]辽宁医学院武警后勤学院附属医院研究生培养基地,天津300162 [2]中国人民武装警察部队后勤学院附属医院心脏中心心血管病研究所天津市心血管重塑与靶器官损伤重点实验室,天津300162 [3]中国人民武装警察部队后勤学院病原生物与免疫学教研室,天津300309

出　　处：《细胞与分子免疫学杂志》2016年第5期625-629,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81570335);天津市应用基础与前沿技术研究计划(14JCZDJC36700)

摘　　要：目的初步探讨土荆皮乙酸(PLAB)对脂多糖(LPS)诱导RAW264.7细胞抗炎作用及影响M1表型偏移的分子机制。方法 LPS诱导RAW264.7细胞建立体外炎症模型,给予0.5μmol/L PLAB和1μmol/L过氧化物酶体增殖物激活受体γ(PPARγ)阻滞剂GW9662处理。流式细胞术检测细胞周期变化,实时定量PCR检测PPARγ和M1型巨噬细胞标志物白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)的mRNA表达,Western blot法检测核因子κB(NF-κB)信号通路相关信号分子水平。结果 PLAB能够明显降低LPS诱导RAW264.7细胞的IL-1β、TNF-α的mRNA水平,上调PPARγ的mRNA水平。下调NF-κB p65、p NF-κB p65、IKKα、IKKβ、p IKKα/β、IκBα、p IκBα的蛋白水平,使RAW264.7细胞阻滞在G0和G2期。GW9662可以抵抗PLAB的抗炎作用。结论 PLAB抑制LPS诱导RAW264.7细胞炎症反应并抑制巨噬细胞向M1表型偏移,与影响细胞周期分布、调控NF-κB/PPARγ通路有关。Objective To investigate the effects of pseudolaric acid B （PLAB） on the inflammatory response and M1 phenotype polarization in RAW264.7 cells induced by lipopelysaccharide （LPS） and the related mechanisms. Methods The inflammatory model in vitro was made using RAW264.7 cells stimulated by LPS, and then was treated with 0.5 μmol/L PLAB and 1 μmol/L GW9662, a peroxisome proliferaters-activated receptor γ （PPARγ） antagonist. The cell cycle was tested by flow cytometry. The mRNA expressions of PPAR-γ and M1 phenotype markers interleukin 16 （IL-1β）, tumor necrosis factor α （TNF-α） were measured by real-time PCR. The expression levels of signal molecules involved in nuclear factor-κB （NF-κB） signal pathway were detected by Western blotting. Results PLAB markedly decreased the expressions of IL-1β and TNF-α mRNAs induced by LPS and increased PPAR）, mRNA level. Moreover, the expressions of NF-κB 1065, pNF-κN p65,IKKα, IKKβ, pIKKα/β, IKBα and pIκBα decreased in PLAB-treated cells. Meanwhile, RAW264.7 cells were arrested in GO and G2 phase after the treatment with PLAB. However, the effects of PLAB on RAW264.7 cells could be reversed by GW9662 obviously. Conclusion PLAB could inhibit the inflammatory response and M1 phenotype polarization in RAW264.7 cells induced by LPS via modulating cell cycle and NF-κB/PPARγ signal pathway.

关 键 词：土槿乙酸 RAW264.7细胞 过氧化物酶体增殖物激活受体γ(PPARγ) 核因子κB(NF-κB) 炎症反应 

分 类 号：R285.5[医药卫生—中药学] R392.12[医药卫生—中医学]