扶正化瘀方对脂多糖诱导的RAW264.7巨噬细胞M1型炎性极化JNK通路的影响(1)

http://www.100md.com 2020年3月1日 《中国中医药信息杂志》 20203

     摘要：目的 觀察扶正化瘀方对脂多糖(LPS)诱导的M1型RAW264.7巨噬细胞一氧化氮合酶(iNOS)基因和一氧化氮(NO)分子过表达的影响，探讨其可能的抗炎机制。方法 采用LPS刺激小鼠RAW264.7巨噬细胞，建立M1型巨噬细胞炎性细胞模型，分别给予扶正化瘀方50、100、200 μg/mL和10 nmol/L JNK蛋白抑制剂SP600125进行孵育。Griess法检测炎症介质NO含量，RT-PCR检测炎症因子白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和iNOS基因表达，Western blot检测iNOS蛋白表达和MAPK信号通路中JNK、p38、ERK蛋白的磷酸化水平。结果 与LPS比较，扶正化瘀方和SP600125均显著降低巨噬细胞中NO分泌(P<0.01)和iNOS蛋白表达(P<0.01)，降低MAPK信号通路JNK蛋白的磷酸化水平(P<0.05，P<0.01);扶正化瘀方对炎症因子IL-6、TNF-α的基因表达及MAPK信号通路ERK、p38蛋白的磷酸化水平无明显影响。结论 扶正化瘀方可能通过抑制JNK蛋白的磷酸化而抑制iNOS基因过表达，最终减少NO的生成，从而抑制RAW264.7巨噬细胞M1型极化，发挥抗炎作用。
, 百拇医药
    关键词：扶正化瘀方;巨噬细胞;一氧化氮;一氧化氮合酶;MAPK信号通路;JNK信号通路

    中图分类号：R285.5 文献标识码：A 文章编号：1005-5304(2020)03-0043-05

    DOI：10.3969/j.issn.1005-5304.201909314

Effects of Fuzheng Huayu Prescription on LPS-induced Inflammatory M1 Polarization of RAW264.7 Macrophages Through JNK Pathway

    ZHANG Man1， HU Xudong2， HUANG Kai1， TAO Yanyan1， PENG Yuan1， LIU Chenghai1，3，4
, 百拇医药
    1. Institute of Liver Diseases， Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine， Shanghai 201203， China; 2. School of Basic Medical Sciences， Shanghai University of Traditional Chinese Medicine， Shanghai 201203， China; 3. Key Laboratory of Liver and Kidney Diseases of Shanghai University of Traditional Chinese Medicine， Ministry of Education， Shanghai 201203， China; 4. Shanghai Key Laboratory of Clinical Chinese Medicine， Shanghai 201203， China
, 百拇医药
    Abstract： Objective To explore the effects of Fuzheng Huayu Prescription on iNOS gene overexpression and NO overproduction in M1 polarized RAW264.7 macrophages induced by lipopolysaccharide (LPS); To discuss its possible anti-inflammatory mechanism. Methods LPS was used to stimulate mouse RAW264.7 macrophages to establish an inflammatory M1 macrophage model， and Fuzheng Huayu Prescription (50， 100， 200 μg/mL) and 10 nmol/L JNK inhibitor SP600125 were incubated respectively. Inflammatory mediator NO production was measured by Griess method. The mRNA expressions of inflammatory cytokines IL-6， TNF-α and iNOS were detected by Real-time RT-PCR. iNOS protein expression and the phosphorylation levels of key proteins in MAPK signaling pathway， namely p-JNK， p-p38 and p-ERK were detected by Western blot. Results Compared with LPS， Fuzheng Huayu Prescription and SP600125 could significantly reduce the overproduction of NO (P<0.01)， the overexpression of, 百拇医药(张满 胡旭东 黄恺 陶艳艳 彭渊 刘成海)

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