原肌球蛋白1在大鼠肝纤维化模型及肝星状细胞中的动态表达及其意义  

Dynamic expression of tropomyosinl in rat model of hepatic fibrosis and hepatic stellate cells

作　　者：刘俊[1] 王春莹[1] 陈永平[1] 林镯[1] 阳韬[1] 陆小蒟[1] 

机构地区：[1]温州医学院附属第一医院感染内科,325000

出　　处：《中华肝脏病杂志》2011年第11期848-852,共5页Chinese Journal of Hepatology

基　　金：王宝恩肝纤维化研究基金（20080017）;浙江省自然科学基金（Y207464）

摘　　要：目的观察原肌球蛋白1（TPMl）在大鼠肝纤维化模型及肝星状细胞（HSC）中的动态表达。方法将SD大鼠随机分为正常对照组（6只）和模型组（24只）。二甲基亚硝胺腹腔注射建立大鼠肝纤维化模型，于第2、4、6、8周（每组各6只）门静脉采血及取肝组织标本；HSC-T6细胞设对照组及刺激组，刺激组以5ng／ml转化生长因子βl（TGFβ1）作用48h。苏木素－伊红和Masson染色观察肝组织病理变化，RT-PCR、免疫组织化学和Westemblot检测组织与细胞中TPMl，TGFβ1及α平滑肌肌动蛋白（α-SMA）的mRNA和蛋白的表达，以及TPMl在肝组织中的定位。两样本均数比较采用独立样本t检验；相关性采用Pearson直线相关分析。结果成功建立肝纤维化模型，TPMl在正常肝组织中低表达于汇管区血管内皮上，在模型组TPMl强表达于增生的肝纤维间隔，TPMl及α－SMA的mRNA及蛋白的表达在肝纤维化过程中均逐渐升高，6周时高于其他各组，8周时下降，与对照组相比，差异均有统计学意义（P〈0．05）；TGFβ1先升高，4周时高于其他各组，6周时下降（P〈0．05）；相关性分析表明TPMl与α－SMA和TGFβ1的表达均呈正相关（rs=0．688和rs=0．692，P〈0．01）；HSC－T6细胞中，TGFβ1刺激组TPMl及α－SMA的mRNA表达均升高，差异有统计学意义（P〈0．05）。结论TPMl参与了肝纤维化的发生和发展过程，有望成为肝纤维化诊断与治疗的新靶点。Objective To investigate the dynamic expression of TPM1 in rat model of hepatic fibrosis and hepatic stellate cells induced by TGFβ1. Method Thirty male SD rats were divided into control group （n = 6） and model group （n = 24）. The rat model of hepatic fibrosis was established by intraperitoneal injection of dimethylnitrosamine（DMN）, the sera were collected from portal vein and liver tissues were taken from animals 2,4,6,8 weeks HSC-T6 cells were cutured and induced 48 hours by 5 ng/ml TGF -β1. The pathological changes of liver were observed by Hematoxylin-Eosin and Masson Staining. Reverse Transcription-polymerase chain reaction （RT-PCR）, immunohistochemistry and Western-blotting were used to determine the mRNA and protein expressions of TPM1, TGFβ1and α-SMA in rat models and HSC-T6 cells and the localization of TPM1 in rat models. Results rat models of hepatic fibrosis were successfully established. TPM1 was lowly stained in the wall of blood vessels in portal areas in normal livers, in fibrotic livers TPM1 was mainly stained along the fibrotic septum. The mRNA and protein expressions of TPM1 and ct-SMA in rat models of hepatic fibrosis increased at the week 2 and peaked at week 6,, which was statistical significance compared to control group, P 〈 0,05; TGF-β1 increased at week 2 and it was higher than the levels in other groups at week 4, which was statistical significance compared to control groupt P 〈 0.05; Correlation analysis showed that TPM1 positively correlated with a-SMA and TGF-β1, rs = 0.688, rs = 0.692, P 〈 0.01. In HSC-T6, the mRNA expressions of TPM1 and α-SMA increased after being induced by TGF -β1. compare with control group, the differences were significant, P 〈 0.05. Conclusion TPM1 may be playing an important role in the occurrence and development of liver fibrosis. Maybe it could become a potential therapeutic target for hepatic fibrosis.

关 键 词：肝硬化 原肌球蛋白 转化生长因子Β Α平滑肌肌动蛋白 

分 类 号：R575.2[医药卫生—消化系统]