Adenoviral expression of a transforming growth factor-beta1 antisense mRNA is effective in preventing liver fibrosis in bile-duct ligated rats.
Adenoviral expression of a transforming growth factor-beta1 antisense mRNA is effective in preventing liver fibrosis in bile-duct ligated rats.
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DOI：
10.1186/1471-230x-3-29
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发表时间：
2003-10-18
影响因子：
2.4
通讯作者：
Weiskirchen R
中科院分区：
文献类型：
--
作者：
Arias M;Sauer-Lehnen S;Treptau J;Janoschek N;Theuerkauf I;Buettner R;Gressner AM;Weiskirchen R
转化生长因子-β(转化生长因子-β)是肝纤维化形成过程中的关键介质。因此，转化生长因子-β作为致病因子可能成为抗肝纤维化基因治疗的主要靶点。我们先前已经证明，通过腺病毒载体携带结构性表达转化生长因子-β1反义基因的转基因可以阻断培养激活的肝星状细胞转化生长因子-β的合成，并有效地消除体外正在进行的纤维化形成。结扎胆总管复制大鼠肝纤维化模型。在此肝损伤模型上，分析转化生长因子-β-1反义核酸对肝纤维化形成的影响。在目前的研究中，我们证明了腺病毒载体指导的mRNA量的合成大约是内源性转化生长因子-β1mRNA量的8000倍。在结扎胆总管引起的实验性损伤大鼠肝脏中，腺病毒载体的给药会抑制转化生长因子-β促进的胶原和α-平滑肌肌动蛋白的产生。此外，应用Ad5-CMV-AS-α-β1后，由于激活的肝星状细胞在胆管结构周围的活跃募集而导致的转化生长因子-平滑肌肌动蛋白阳性细胞的数量显著减少。然而，在该阻塞性肝损伤模型中观察到的升高的血清天冬氨酸转氨酶、丙氨酸转氨酶和胆红素水平在转化生长因子-β拮抗剂的存在下没有显著改变。综上所述，我们的数据提供了体内证据，即转化生长因子-β1反义基因的传递特异性地取消了直接转化生长因子-β功能在持续肝纤维化中的各种作用。因此，我们得出结论，表达的转基因对于抑制转化生长因子-β的作用在治疗上是有用的，从阐明转化生长因子-β在肝损伤过程中的功能到开发新的基因治疗方法。
Transforming growth factor-β (TGF-β) is a key mediator in establishing liver fibrosis. Therefore, TGF-β as a causative agent may serve as a primary target for antifibrotic gene therapy approaches. We have previously shown that the adenoviral delivery of a transgene constitutively expressing a TGF-β1 antisense mRNA blocks TGF-β synthesis in culture-activated hepatic stellate cells and effectively abolishes ongoing fibrogenesis in vitro. Ligature of the common bile duct was used to induce liver fibrosis in rats. The effect of the TGF-β1 antisense on fibrogenesis was analyzed in this model of liver injury. In the present study, we demonstrate that the adenoviral vector directs the synthesis of mRNA quantities that are approximately 8000-fold more abundant than endogenous TGF-β1 mRNA. In experimentally injured rat livers induced by ligature of the common bile duct, a model for persistent fibrogenesis and cirrhosis, administration of the adenoviral vector abrogates TGF-β-enhanced production of collagen and α-smooth muscle actin. Furthermore, the number of cells positive for α-smooth muscle actin resulting from active recruitment of activated hepatic stellate cells around the bile ductular structures was significantly reduced in animals after application of Ad5-CMV-AS-TGF-β1. However, the observed elevated serum levels of aspartate aminotransferase, alanine aminotransferase, and bilirubin induced in this obstructive liver injury model were not significantly altered in the presence of the TGF-β antagonist. Taken together, our data provides in vivo evidence that the delivery of TGF-β1 antisense mRNA specifically abolishes the diverse effects of direct TGF-β function in ongoing liver fibrogenesis. Therefore, we conclude that the expressed transgene is therapeutically useful for inhibition of TGF-β effects in diverse applications, ranging from clarification of TGF-β function in the course of liver injury to the development of novel gene therapeutic approaches.