摘要
目的:探讨卫矛粗提物对M1型巨噬细胞的作用及其作用机制。方法:采用Griess法、免疫酶联吸附试验(ELISA)、实时荧光定量聚合酶链反应(q RT-PCR)、流式细胞术和免疫印迹(Western Blot)法考察卫矛粗提物对脂多糖(LPS)建立的M1亚型巨噬细胞模型的作用及机制。结果:1μg/m L的LPS诱导巨噬细胞12h后,可成功构建M1巨噬细胞模型;与LPS模型组比较,卫矛粗提物组可以明显抑制巨噬细胞分泌NO、白细胞介素(IL)-6和肿瘤坏死因子α(TNF-α)以及下调i NOS、IL-6和TNF-αm RNA的表达水平,降低M1型巨噬细胞膜表面分子CD16/32的阳性表达率,并对磷酸化的氨基末端激酶(JNK)和细胞外信号调节激酶(ERK1/2)蛋白表达具有明显抑制作用(P<0.05)。结论:卫矛粗提物对M1型巨噬细胞具有良好的抑制作用,可能与其抑制丝裂原活化蛋白激酶(MAPK)信号转导通路中JNK和ERK1/2蛋白的磷酸化有关。
Objective: To investigate the effects and mechanisms of the crude extract of Euonymus alatus on M1 macrophage. Methods: The effects and mechanisms of the crude extract of Euonymus alatus on M1 macrophage was inspected by was established by lipopolysaccharide(LPS) through the methods such as Griess assay, enzyme-linked immunosorbent assay(ELISA), real-time quantitative PCR(q RT-PCR), flow cytometry and the Western Blot. Results: The cell model of M1 macrophage was successfully established by treating with LPS for 12 h. Compare with the LPS group, the crude extract of Euonymus alatus group could inhibit the M1 macrophage secret nitric oxide(NO), interleukin 6(IL-6) and tumor necrosis factor(TNF-α), decrease the m RNA expression of inducible nitric oxide synthase(i NOS), IL-6 and TNF-α m RNA, reduce the positive expression rate of the CD16/32 that was a cell membrane surface molecule of M1 macrophage, and obviously restrain the protein expression of c-jun N-terminal kinase(JNK) in phosphorylation and extracellular signal-regulated kinase(ERK1/2)(P<0.05). Conclusion: The crude extract of Euonymus alatus can effectively inhibit the polarization of M1 macrophage, which is related to inhibit the protein phosphorylation of JNK and ERK1/2 in MAPK signaling pathway.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2017年第10期4407-4411,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国医大师洪广祥全国名老中医药专家传承工作室[国中医药发(2014)20号]
国家自然科学基金项目(No.81460708
No.81560765
No.81660760)
江西省高等学校科技落地计划项目(No.KJLD13063)
江西省自然科学基金项目(No.20132BAB205093)
江西省卫生厅中医药科研计划项目(No.2013Z001)
江西省教育厅科研项目(No.GJJ14600)~~
关键词
卫矛粗提物
M1型巨噬细胞
细胞因子
信号通路
Crude extract of Euonymus alatus
M1 macrophage
Cytokines
Signaling pathway
