發(fā)布時(shí)間：2018-04-29 07:07

  本文選題：骨髓細(xì)胞 + 表沒食子兒茶素-3-沒食子酸酯��； 參考：《河南師范大學(xué)》2017年碩士論文

【摘要】：巨噬細(xì)胞是一類具有高度異質(zhì)性和可塑性的免疫細(xì)胞群體,起源于骨髓的單核細(xì)胞,能夠改變自己的表型以適應(yīng)于所處微環(huán)境。巨噬細(xì)胞極化分為經(jīng)典活化型巨噬細(xì)胞(classically activated macrophage,M1)和替代活化型巨噬細(xì)胞(alternativelly activated macrophage,M2)兩種類型。巨噬細(xì)胞在γ-干擾素(IFN-γ)、細(xì)菌脂多糖(LPS)刺激下分化為M1型巨噬細(xì)胞,分泌多種促炎細(xì)胞因子、活性氧等,如TNF-α、IL-1β、ROS等以清除病原體、壞死組織和激活其他免疫細(xì)胞,幫助機(jī)體抵御感染。M2型巨噬細(xì)胞由IL-4、IL-13誘導(dǎo)產(chǎn)生,可分泌多種抗炎因子,如Ym-1、IL-10等,主要出現(xiàn)在炎癥消退階段有助于組織修復(fù)。表沒食子兒茶素沒食子酸酯(epigallocatechin-3-gallate,EGCG)是綠茶多酚的主要活性成分,具有抗氧化、抗炎、抗癌等多種生物學(xué)效應(yīng)。EGCG抗癌和抗炎作用機(jī)制已有較多的研究。但EGCG能否影響髓源性巨噬細(xì)胞的極化,目前并不清楚。因此,本實(shí)驗(yàn)通過觀察EGCG對(duì)髓源性巨噬細(xì)胞分化的M1型巨噬細(xì)胞和M2型巨噬細(xì)胞的標(biāo)志性因子的表達(dá)情況探討EGCG對(duì)髓源性巨噬細(xì)胞極化的影響。本實(shí)驗(yàn)以6~8周齡C57BL/6小鼠骨髓細(xì)胞為材料,經(jīng)100 ng/mL巨噬細(xì)胞集落刺激因子(macrophage colony-stimulating factor,M-CSF)體外誘導(dǎo)分化為巨噬細(xì)胞(bone marrow-derived macrophages,BMDM),加入50 ng/mL的IFN-γ和1μg/m L的LPS刺激分化為M1型巨噬細(xì)胞,或者加入20 ng/mL的IL-4刺激分化為M2型巨噬細(xì)胞,然后暴露于12.5~50μmol/L的EGCG處理,7天后收集細(xì)胞提取RNA和蛋白,利用實(shí)時(shí)熒光定量PCR(RT-PCR)和酶聯(lián)免疫吸附試驗(yàn)(ELISA),檢測(cè)M1型巨噬細(xì)胞標(biāo)志物i NOS和促炎因子IL-1β、TNF-α以及M2型巨噬細(xì)胞標(biāo)志物Arg-1、Ym-1和抗炎因子IL-10的mRNA表達(dá)水平和蛋白含量。結(jié)果顯示,IFN-γ和LPS刺激使髓源性巨噬細(xì)胞的IL-1β、TNF-α和iNOS mRNA和蛋白水平上調(diào),而EGCG處理可抑制IFN-γ和LPS刺激的IL-1β、TNF-α和iNOS表達(dá),且具有劑量依賴效應(yīng)。IL-4刺激使髓源性巨噬細(xì)胞的Arg-1、Ym-1、IL-10表達(dá)上調(diào),EGCG處理組可增強(qiáng)Arg-1、Ym-1、IL-10的表達(dá),且存在劑量依賴效應(yīng)。上述結(jié)果表明,EGCG以劑量依賴方式減弱M1型巨噬細(xì)胞標(biāo)志因子的表達(dá),而增強(qiáng)M2型巨噬細(xì)胞標(biāo)志因子的表達(dá)。
[Abstract]:Macrophages are a class of immune cells with high heterogeneity and plasticity. They are derived from bone marrow mononuclear cells and can change their phenotypes to adapt to their microenvironment. The polarization of macrophages can be divided into two types: classical activated activated macrophage M _ 1) and alternative activated activated macrophage M _ 2). Macrophages, stimulated by IFN- 緯, lipopolysaccharide (LPS), differentiate into M1 macrophages, secrete a variety of pro-inflammatory cytokines and reactive oxygen species, such as TNF- 偽, IL-1 尾 Ros, in order to clear pathogens, necrotic tissue and activate other immune cells. The macrophages can be induced by IL-4 and IL-13, and secrete many kinds of anti-inflammatory factors, such as Ym-1, IL-10 and so on. Epigallocatechin-3-gallate EGCGG is the main active component of green tea polyphenols, and has many biological effects, such as anti-oxidation, anti-inflammation, anti-cancer and so on. The mechanism of anti-cancer and anti-inflammatory effects of EGCG has been studied. However, it is not clear whether EGCG can affect the polarization of myelogenous macrophages. Therefore, the effect of EGCG on the polarization of myelogenous macrophages was investigated by observing the expression of the marked factors of M1 type macrophages and M2 type macrophages of myelogenous macrophages. In this study, bone marrow cells of C57BL/6 mice aged 6 to 8 weeks were used as materials. Macrophage colony-stimulating factor M-CSF (100 ng/mL macrophage colony-stimulating factor) was used to induce the differentiation of macrophages into marrow-derived macrophages BMDM, and 50 ng/mL IFN- 緯 and 1 渭 g / mL LPS were added to induce macrophages to differentiate into M1 macrophages. Alternatively, 20 ng/mL IL-4 was added to stimulate differentiation into M2 macrophages, and then exposed to 12.5 渭 mol/L EGCG for 7 days, the cells were collected to extract RNA and protein. The mRNA expression and protein content of M 1 macrophage markers I NOS, IL-1 尾 TNF- 偽, M 2 macrophage markers Arg-1Tm-1 and anti-inflammatory factor IL-10 were detected by real-time fluorescence quantitative PCR and Elisa assay. The results showed that IFN- 緯 and LPS increased the levels of IL-1 尾 -TNF- 偽 and iNOS mRNA and protein in myelogenous macrophages, while EGCG treatment inhibited the expression of IL-1 尾 -TNF- 偽 and iNOS stimulated by IFN- 緯 and LPS. The expression of Ym-10 in myelogenous macrophages increased in a dose-dependent manner after stimulation with IL-4. In EGCG treated group, the expression of IL-10 was enhanced in a dose-dependent manner, and there was a dose-dependent effect on the expression of IL-10. These results suggested that EGCG could attenuate the expression of M1 type macrophage marker factor in a dose-dependent manner, but enhance the expression of M 2 type macrophage marker factor.
【學(xué)位授予單位】：河南師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)碩士學(xué)位論文 前1條

1 劉敏;表沒食子兒茶素沒食子酸酯對(duì)白介素-4誘導(dǎo)的小鼠M2型巨噬細(xì)胞標(biāo)志性因子表達(dá)的影響[D];河南師范大學(xué);2016年

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本文編號(hào)：1818911