不同铁负载水平影响肝星状细胞活化和凋亡的抗肝纤维化机制研究  

An anti-fibrosis mechanism study on effects of difference in iron load levels on activation and apoptosis of hepatic stellate cells

作　　者：江远[1] 张玲[1] 邓远绮 何金洋[2] 

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

出　　处：《中国中西医结合急救杂志》2013年第6期369-373,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：基金项目：广东省中医药管理局项目（20122017）

摘　　要：目的：探讨不同铁负载水平影响肝星状细胞（HSC）活化和凋亡的抗肝纤维化机制。方法根据细胞内不同铁负载水平将肝星状细胞株（HSC-T6细胞）分为空白对照组、铁沉积模型组、50μmol/L去铁铵组和25μmol/L去铁铵组共4组。用定量聚合酶链反应（PCR）检测HSC-T6细胞Ⅰ型胶原和转化生长因子-β1（TGF-β1）的mRNA表达；免疫组化法检测HSC-T6细胞α-平滑肌肌动蛋白（α-SMA）的表达；原位末端缺刻标记法（TUNEL）检测HSC-T6细胞的凋亡；电镜下观察HSC-T6细胞超微结构。结果与空白对照组比较，铁沉积模型组TGFβ1 mRNA表达虽有所增强，但差异无统计学意义（1.594±0.168比1.477±0.126，P＞0.05），Ⅰ型胶原mRNA表达则显著增强（1.354±0.076比1.197±0.104，P＜0.01）。50μmol/L和25μmol/L去铁铵均能下调Ⅰ型胶原和TGF-β1的mRNA表达，且50μmol/L去铁铵组优于25μmol/L去铁铵组（Ⅰ型胶原mRNA：0.391±0.076比0.688±0.060，TGF-β1 mRNA：0.421±0.068比0.714±0.090，均P＜0.01）。铁沉积可诱导HSC中α-SMA大量表达，却仅见有较少的凋亡细胞。而经去铁铵处理后HSC表达α-SMA明显减少，但HSC发生了凋亡。结论细胞内不同铁负载水平能够诱导HSC活化或凋亡，表明铁在调节HSC活化和凋亡过程中发挥重要作用，而且也揭示了铁螯合剂治疗肝纤维化的潜在作用。Objective To investigate the anti-fibrosis mechanism from effects of difference in iron load levels on activation and apoptosis of hepatic stellate cells（HSCs）. Methods According to the difference in iron load levels in HSCs,HSC-T6 cells were divided into four groups：blank control,iron deposition model,50μmol/L desferrioxamine and 25 μmol/L desferrioxamine groups. Quantitative polymerase chain reaction（PCR）was applied for the detection of collagen type Ⅰ and transforming growth factor-β1（TGF-β1）mRNA expressions of HSC-T6 cells. Immunohistochemical assay was used for the detection of α-smooth muscle actin（α-SMA）expression. The method of terminal deoxynucleotidyl transferase（TdT）-mediated dUTP nick end labeling（TUNEL）was used for the examination of apoptosis of HSC-T6 cells. Under electron microscope,the ultrastructures of HSC-T6 cells were observed. Results Compared with the blank control group,despite the TGF-β1 mRNA expression in iron deposition model group was increased,no statistical significant difference was seen（1.594±0.168 vs. 1.477±0.126, P〉0.05）,whereas collagen type I mRNA expression was significantly enhanced（1.354±0.076 vs. 1.197±0.104, P〈0.01）. Both 50μmol/L and 25μmol/L desferrioxamine could down-regulate collagen type I and TGF-β1 mRNA expressions,and the action of 50μmol/L desferrioxamine was superior to that of 25μmol/L desferrioxamine（collagen typeⅠmRNA：0.391±0.076 vs. 0.688±0.060,TGF-β1 mRNA：0.421±0.068 vs. 0.714±0.090,both P〈0.01）. Iron deposition could induce HSCs to expressα-SMA in great amount,while apoptosis could be seen scarcely in iron deposited HSCs. By desferrioxamine therapy,α-SMA expression of HSCs was decreased significantly,but some of the cells underwent apoptosis. Conclusion Different iron load levels inside HSCs can induce activation or apoptosis of the cells,showing that iron plays an important role in regulating the process of HSCs activation and apoptosis and revealing that desferrioxamine possesses t

关 键 词：肝星状细胞 肝纤维化 铁 去铁铵 活化 凋亡 

分 类 号：R575.2[医药卫生—消化系统]