本文選題：慢性丙型肝炎　切入點(diǎn)：鐵過(guò)載　出處：《細(xì)胞與分子免疫學(xué)雜志》2017年08期

【摘要】：目的觀察鐵過(guò)載對(duì)人肝癌Huh7.5細(xì)胞活性的影響及機(jī)制。方法用(50、100、200)μmol/L枸櫞酸鐵銨(FAC)處理Huh7.5細(xì)胞,鐵離子熒光探針Phen Green FL標(biāo)記結(jié)合熒光顯微鏡檢測(cè)細(xì)胞鐵載量。MTT法檢測(cè)鐵過(guò)載細(xì)胞的增殖活性,實(shí)時(shí)定量PCR檢測(cè)鐵過(guò)載細(xì)胞運(yùn)鐵蛋白受體1(TfR1)、TfR2、二價(jià)金屬離子轉(zhuǎn)運(yùn)體1(DMT1)和膜鐵轉(zhuǎn)運(yùn)蛋白1(FPN1)的mRNA水平,Western blot法檢測(cè)TfR1、TfR2、DMT1和FPN 1蛋白水平,二氯二氫熒光素乙酰乙酸酯(DCFH-DA)染色結(jié)合流式細(xì)胞術(shù)檢測(cè)活性氧(ROS)水平;在FAC處理細(xì)胞以及FAC聯(lián)合400μmol/L N-乙酰半胱氨酸(NAC)處理后,采用異硫氰酸熒光素標(biāo)記的膜聯(lián)素Ⅴ/碘化丙啶(annexinⅤ-FITC/PI)雙染色結(jié)合流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡。結(jié)果細(xì)胞鐵載量隨FAC增加呈濃度依賴性上升;FAC處理細(xì)胞TfR1、TfR2和DMT1 mRNA和蛋白水平顯著下調(diào),FPN1 mRNA和蛋白水平顯著上調(diào);ROS水平隨FAC濃度增加顯著升高,細(xì)胞增殖活性隨FAC濃度增加顯著下降;FAC處理細(xì)胞的凋亡率均顯著高于對(duì)照組,而向FAC處理細(xì)胞加入抗氧化劑NAC后,細(xì)胞凋亡率顯著下降。結(jié)論鐵過(guò)載抑制Huh7.5細(xì)胞增殖活性并促進(jìn)細(xì)胞凋亡,其作用機(jī)制可能與氧化應(yīng)激途徑有關(guān)。
[Abstract]:Objective to observe the effect of iron overload on the activity of human hepatoma Huh7.5 cells and its mechanism. Methods Huh7.5 cells were treated with 50 100 200 渭 mol/L ferric ammonium citrate (FAC). The ferric ion fluorescence probe Phen Green FL labeling and fluorescence microscope were used to detect the iron load of the cells and the proliferation activity of the iron overload cells. Real-time quantitative PCR was used to detect the levels of TfR2, divalent metal ion transporter 1 (DMT1) and membrane iron transporter 1 (FPN1) in iron-overload cells. Western blot was used to detect the levels of TfR1TfR2DMT1 and FPN 1 protein. DCFH-DAA staining combined with flow cytometry (FCM) was used to detect the level of reactive oxygen species (Ros) in cells treated with FAC and FAC combined with 400 渭 mol/L N-acetylcysteine (NAC). Apoptosis was detected by double staining of annexin 鈪，

本文編號(hào)：1679792