铁沉积对大鼠肝纤维化影响的机制研究  

Effects of iron deposition on liver fibrosis in rats

作　　者：江远[1] 张玲[1] 钟肖英[1] 吴文如[2] 何金洋[2] 

机构地区：[1]广州医科大学附属深圳沙井医院中医肝病科,广东省深圳市518104 [2]广州中医药大学

出　　处：《实用肝脏病杂志》2015年第2期173-177,共5页Journal of Practical Hepatology

基　　金：广东省中医药管理局项目(项目编号:20122017)

摘　　要：目的探讨铁沉积对大鼠肝纤维化影响的机制。方法随机将39只SD大鼠分为模型组和空白对照组,采用二甲基亚硝胺(DMN,10μL·kg-1)腹腔注射,制作大鼠肝纤维化模型。造模大鼠在注射DMN 1 w后,再将模型大鼠随机分为模型组(15只)和去铁铵组(12只)。两组大鼠分别自第3 w开始腹腔注射生理盐水或100 mg·kg-1去铁铵,3次/w,2 w后处死动物。取肝组织分别行HE染色、Masson染色、普鲁士蓝染色;采用免疫组化法检测肝组织α-平滑肌肌动蛋白(α-SMA)的表达;采用火焰原子吸收光谱法(FAAS)测定大鼠肝组织铁浓度(HIC);采用ELISA法检测大鼠血清铁蛋白、转铁蛋白;使用全自动生化分析仪检测肝功能、血清铁水平;采用PCR法检测肝组织转化生长因子(TGF)-β1 m RNA水平。结果肝组织病理学检查显示,伴随着胶原纤维的沉积、肝细胞变性坏死和肝星状细胞(HSC)大量活化,模型组大鼠铁负载显著增加;铁沿纤维间隔分布,主要沉积于库普弗细胞(KC)和HSC;模型组肝组织铁浓度为(0.778±0.098)mg/g,空白对照组为(0.436±0.043)mg/g,两组差别有统计学意义(LSD-t=5.15,P<0.01);去铁铵组为(0.595±0.146)mg/g,显著低于模型组(LSD-t=-2.76,P<0.05);模型组血清铁蛋白和转铁蛋白分别为(47.657±27.851)ng/m L和(0.322±0.099)mg/m L,空白对照组分别为(24.166±27.626)ng/m L和(0.653±0.170)mg/m L,去铁铵组分别为(10.261±12.466)ng/m L和(0.584±0.180)mg/m L,说明模型组铁蛋白明显增加,血清转铁蛋白明显减少,与空白对照组比较,差异均有统计学意义(LSD-t=2.21和-4.78,P<0.05和P<0.01);去铁铵能明显降低血清铁蛋白水平,增加血清转铁蛋白水平,与模型组比较差异有统计学意义(LSD-t=-3.52和3.77,P<0.05和P=0.01);模型组肝组织TGF-β1m RNA水平为(11.896±0.63),空白对照组为(2.292±0.222),两组差别有统计学意义(LSD-t=25.95,P<0.01),去铁铵组为(7.481±0.745),显著低于模型组(LSD-t=-11.95,P<0.01)。结论铁沉积对Objective To investigate the effects of iron deposition on the liver fibrosis and its potential mechanisms in rats. Methods 39 SD rats were randomly divided into fibrosis group （n=27） and control group （n=12）. Rats with hepatic fibrosis （n=27） were established by intraperitoneal injection of dimethylnitrosamine （DMN,10 μL.kg-1）. After one-week DMN injection,the 27 rats were randomly divided into model group （n=15） and desferrioxamine group （n=12）. At the beginning of the third week,rats in model and desferrioxamine group were respectively injected intraperitoneally with normal saline or desferrioxamine at the dose of 100 mg＆#183;kg-1＆#183;d-1,3 times per week for 2 weeks before they were sacrificed. Liver tissues were stained with HE,Masson and Prussian blue,respectively;Immunohistochemistry was applied for the detection of α-smooth muscle actin （α-SMA） expres-sion;Hepatic iron concentration （HIC） in liver tissues were evaluated by flame atomic absorption spectrophotome-try （FAAS）;ELISA was adopted to examine the concentrations of serum ferritin and transferrin. Automatic bio-chemical analyzer was used to detect the liver func-tion and the serum level of iron. The mRNA levels of transforming growth factor-β1 （TGF-β1） was de-tected by quantitative PCR. Results Our histopatho-logical findings showed that iron loads in liver tissues in model group increased obviously,accompanied with excessive collagen deposition,hepatocyte denaturation and necrosis and activation of a great number of hepatic stellate cells （HSCs） and the iron distributed in fibrous septa with main deposition in Kupffer cells（KCs）and HSCs;The hepatic iron concentration in model group [（0.778± 0.098） mg/g] was higher than that in control group[（0.436±0.043） mg/g,LSD-t=5.15,P〈0.01] and than in desfer-rioxamine group[（0.595±0.146） mg/g,LSD-t=-2.76,P〈0.05];Compared with in control,the level of serum ferritin in model group significantly increased,and the level of serum transferr

关 键 词：肝纤维化 铁 去铁铵 肝星状细胞 库普弗细胞 大鼠 

分 类 号：R575.2[医药卫生—消化系统]