Adenovirus-mediated inhibition of SPARC attenuates liver fibrosis in rats
Adenovirus-mediated inhibition of SPARC attenuates liver fibrosis in rats
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DOI：
10.1002/jgm.1228
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发表时间：
2008-09-01
影响因子：
3.5
通讯作者：
Mazzolini, Guillermo
中科院分区：
文献类型：
--
作者：
Camino, Alejandra M.;Atorrasagasti, Catalina;Mazzolini, Guillermo
背景肝纤维化发生过程中，纤维化细胞与细胞外基质之间的相互作用发挥了重要作用，但其机制尚不清楚。分泌蛋白，酸性和富含半胱氨酸（Cys）是一种基质细胞糖蛋白，由肝星状细胞表达，并在纤维化肝脏中过度表达。方法构建携带反义β-淀粉样蛋白基因的重组腺病毒Adasaplase，并将其转染大鼠肝纤维化模型，观察反义β-淀粉样蛋白在肝纤维化中的作用。在Sprague-Dawley大鼠中通过长期腹腔内给予硫代乙酰胺诱导晚期肝纤维化。在第一次给药后1周，动物通过尾静脉接受Adaslavin或Ad β gal（对照腺病毒）的注射并直接注射到肝脏中。8周后观察肝组织病理学改变及肝纤维化指标。采用实时定量聚合酶链反应、免疫印迹、酶联免疫吸附试验和免疫组织化学方法检测肝纤维化大鼠肝组织中TGF-β和α-平滑肌肌动蛋白的表达。通过胶原沉积减少、肝羟脯氨酸含量降低和纤维化形态学分期较低评估，阿达木单抗显著减弱硫代乙酰胺处理大鼠的肝纤维化发展。阿达木单抗治疗降低了炎症活性（Knodell评分），并在体内抑制了肝星状细胞向肌成纤维细胞样表型的转分化。此外，在体外抑制肝星状细胞的TGF-β的产生减少。结论这是第一次研究表明，敲低肝TGF-β的表达改善硫代乙酰胺诱导的肝纤维化大鼠慢性肝损伤。结论：hepatocellular carcinoma是肝纤维化基因治疗的潜在靶点。版权所有（C）2008约翰威利父子有限公司
Background The interaction between fibrogenic cells and extracellular matrix plays a role in liver fibrosis, yet the mechanisms are largely unknown. Secreted protein, acidic and rich in cysteine (SPARC) is a matricellular glycoprotein that is expressed by hepatic stellate cells and is overexpressed in fibrotic livers. We investigated the in vivo role of SPARC in experimentally induced liver fibrosis in rats.Methods A recombinant adenovirus carrying antisense SPARC was constructed (AdasSPARC). Advanced liver fibrosis was induced in Sprague-Dawley rats by prolonged intraperitoneal administration of thioacetamide. Animals received injections of AdasSPARC or Ad beta gal (control adenovirus) via the tail vein and directly into the liver 1 week after the first dose. The pathological changes in liver tissues and indices of fibrosis were assessed at eight weeks. Expression of SPARC, transforming growth factor (TGF)-beta and a-smooth muscle actin were evaluated by quantitative real-time polymerase chain reaction, western blotting, enzyme-linked immunosorbent assay and immunohistochemistry.Results Hepatic SPARC expression significantly increased during the development of liver fibrosis. AdasSPARC markedly attenuated the development of hepatic fibrosis in rats treated with thiocetamide, as assessed by decreased Collagen deposition, lower hepatic content of hydroxyproline and less advanced morphometric stage of fibrosis. AdasSPARC treatment reduced inflammatory activity (Knodell score) and suppressed transdifferentiation of hepatic stellate cell to the myofibroblasts like phenotype in vivo. Furthermore, in vitro inhibition of SPARC on hepatic stellate cells decreases the production of TGF-beta.Conclusions This is the first study to demonstrate that knockdown of hepatic SPARC expression ameliorates thioacetamide-induced liver fibrosis in rats with chronic liver injury. SPARC is a potential target for gene therapy in liver fibrosis. Copyright (C) 2008 John Wiley & Sons, Ltd.