铁沉积于巨噬细胞对转化生长因子及炎症介质基因表达的影响  

Effects of iron deposition in macrophages on transforming growth factor-β1, inducible nitric oxide synthase and tumor necrosis factor-α mRNA expressions

作　　者：江远[1] 张玲[1] 何金洋[2] 聂广[3] 

机构地区：[1]广州医学院附属深圳沙井医院中医肝病科,广东深圳518104 [2]广州中医药大学热带医学研究所,广东广州510405 [3]深圳市第三人民医院,广东深圳518020

出　　处：《中国中西医结合急救杂志》2011年第2期103-106,I0001,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：广东省深圳市宝安区科技计划项目（2010698）

摘　　要：目的 探讨铁沉积于巨噬细胞对肝纤维化发生发展的作用机制.方法 以柠檬酸铁胺（FAC）与巨噬细胞株（RAW264.7细胞）共同温育,体外建立铁沉积于巨噬细胞模型.①将RAW264.7细胞分为FAC 1 mmol/L、100 μmol/L组及对照组,每组8孔.观察铁沉积于RAW264.7细胞对转化生长因子-β1（TGF-β1）、诱导型一氧化氮合酶（iNOS）、肿瘤坏死因子-α（TNF-α）mRNA表达的影响.②另将RAW264.7细胞分为对照组、铁沉积组、去铁铵组;对照组为正常培养的RAW264.7细胞,铁沉积组加入100 μmol/L FAC,去铁铵组分别加入100 μmol/L FAC和1 000、200、40、8、1.6、0.32 μmol/L的去铁铵,每组3孔.观察不同浓度去铁铵对铁沉积于RAW264.7细胞表达TGF-β1、iNOS、TNF-α mRNA的影响.结果 ①铁沉积于RAW264.7细胞主要使TGF-β1 mRNA表达明显降低,iNOS、TNF-α mRNA表达显著升高,以100 μmol/L FAC作用显著,与对照组比较差异有统计学意义（TGF-β1 mRNA：0.60±0.18比1.42±0.21,iNOS mRNA：3.79±1.48比1.10±0.33,TNF-α mRNA：1.72±0.37比1.07±0.10,P〈0.05或P〈0.01）.②去铁铵能使RAW264.7细胞出现与铁沉积后相反的基因表达变化,以较高浓度1 000 μmol/L、200 μmol/L（TGF-β1 mRNA：3.13±1.03、2.39±0.29,iNOS mRNA：3.28±0.62、5.36±1.04,TNF-α mRNA：3.04±1.35、3.35±0.52）作用显著,与铁沉积组（TGF-β1 mRNA：0.22±0.09,iNOS mRNA：18.57±8.44,TNF-α mRNA：6.30±1.00）比较差异有统计学意义（均P〈0.01）.结论 铁沉积于巨噬细胞可能是通过诱导一氧化氮（NO）过度合成,促进炎症的形成并进一步促进肝纤维化的发生发展,而不是通过上调TGF-β1 mRNA表达来实现的.Objective To investigate the effect of iron deposition in macrophages on the mechanism of liver fibrogenensis. Methods Iron deposited macrophages model was established by incubating with ferric ammonium citrate （FAC） and macrophages line （RAW264.7 cells）.①RAW264.7 cells were divided into 1 mmol/L FAC group, 100μmol/L FAC group and control group, each group repeated for 8 wells. The effects of iron deposition in macrophages on mRNA expressions of transforming growth factor-β1（TGF-β1）, inducible nitric oxide synthase （iNOS） and tumor necrosis factor-α （TNF-α） were investigated. ② Additionally, RAW264.7 cells were divided into control group, iron deposition group and desferrioxamine groups. Control group contained the normally cultured RAW264.7 cells. Iron deposition group was added with 100 μmol/L of FAC. Desferrioxamine groups were differently added with 1 000, 200, 40, 8, 1.6, 0. 32 μmol/L of desferrioxamine and 100 μmol/L of FAC, each group being repeated for 3 wells. The effects of different concentrations of desferrioxamine on mRNA expressions of TGF-β1, iNOS and TNF-α of iron deposited macrophages were investigated. Results ① Iron deposition in macrophages mainly decreased the expression of TGF-β1 mRNA, and increased the mRNA expressions of iNOS and TNF-α. The mRNA expression was more considerable in 100 μmol/L FAC group, and compared with the control group, there were statistical significant differences （TGF-β1 mRNA： 0.60±0.18 vs. 1.42±0.21, iNOS mRNA： 3.79± 1.48 vs. 1.10±0. 33, TNF-a mRNA： 1. 7210.37 vs. 1.07±0.10, P〈0.05 or P〈0.01）.② The opposite gene expression was observed with desferrioxamine therapy, which were more considerable in 1 000 mol/L and 200 μmol/L desferrioxamine groups （TGF-~I mRNA： 3.13±1.03, 2.39±0.29; iNOS mRNA： 3.28± 0.62, 5.36 ± 1.04 ; TNF-α mRNA： 3.04 ± 1.35, 3.35 ± 0.52）. Compared with the iron deposition group （TGF-β1 mRNA： 0.22 ± 0.09, iNOS mRNA： 18.57 ± 8.44, TNF-α mRNA： 6.30 ± 1.00）, the

关 键 词：巨噬细胞 肝纤维化 铁 

分 类 号：R256.4[医药卫生—中医内科学] Q813.11[医药卫生—中医学]