Antifibrotic Effects of a Recombinant Adeno-Associated Virus Carrying Small Interfering RNA Targeting TIMP-1 in Rat Liver Fibrosis
Antifibrotic Effects of a Recombinant Adeno-Associated Virus Carrying Small Interfering RNA Targeting TIMP-1 in Rat Liver Fibrosis
复制标题
携带靶向 TIMP-1 的小干扰 RNA 的重组腺相关病毒对大鼠肝纤维化的抗纤维化作用
DOI：
10.1016/j.ajpath.2013.01.036
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发表时间：
2013-05-01
影响因子：
6
通讯作者：
You, Hong
中科院分区：
文献类型：
--
作者：
Cong, Min;Liu, Tianhui;You, Hong
金属蛋白酶组织抑制剂1（TIMP-1）表达升高有助于肝纤维化细胞外基质的过量产生。本研究构建了携带TIMP-1基因siRNA的重组腺相关病毒（rAAV/siRNA-TIMP-1），并研究了其对大鼠肝纤维化的影响。用rAAV/siRNA-TIMP-1处理两种大鼠肝纤维化模型，四氯化碳和胆管结扎模型。在四氯化碳模型中，rAAV/siRNA-TIMP-1给药减轻了纤维化严重程度，如通过肝胶原积累、羟脯氨酸含量以及肝和血清中I型和III型胶原浓度的组织学分析所确定的。mRNA和活性基质金属蛋白酶（MMP）13的水平升高，而mRNA和活性MMP-2的水平降低。此外，还观察到α-平滑肌肌动蛋白（激活的肝星状细胞（HSC）的生物标志物）和转化生长因子-β 1（对肝纤维化的发展至关重要）的表达显著降低。类似地，rAAV/siRNA-TIMP-1治疗显著减轻胆管结扎诱导的肝纤维化。此外，该处理显著抑制了来自两种模型大鼠的HSC中的TIMP-1表达。这些数据表明，rAAV/siRNA-TIMP-1的施用通过直接提高MMP-13的功能和减少活化的HSC来减轻肝纤维化。它还导致I型胶原、MMP-2和转化生长因子β 1的表达间接降低。结论：rAAV/siRNA-TIMP-1可能是一种有效的抗肝纤维化基因治疗药物。
Elevated tissue inhibitor of metalloproteinase 1 (TIMP-1) expression contributes to excess production of extracellular matrix in liver fibrosis. Herein, we constructed a recombinant adeno-associated virus (rAAV) carrying siRNA of the TIMP-1 gene (rAAV/siRNA-TIMP-1) and investigated its effects on Liver fibrosis in rats. Two models of rat liver fibrosis, the carbon tetrachloride and bile duct ligation models, were treated with rAAV/siRNA-TIMP-1. In the carbon tetrachloride model, rAAV/siRNA-TIMP-1 administration attenuated fibrosis severity, as determined by histologic analysis of hepatic collagen accumulation, hydroxyproline content, and concentrations of types I and III collagen in Livers and sera. Levels of mRNA and active matrix metalloproteinase (MMP) 13 were elevated, whereas levels of mRNA and active MMP-2 were decreased. Moreover, a marked decrease was noted in the expression of alpha-smooth muscle actin, a biomarker of activated hepatic stellate cells (HSCs), and transforming growth factor-beta 1, critical for the development of Liver fibrosis. Similarly, rAAV/siRNA-TIMP-1 treatment significantly alleviated bile duct Ligation-induced liver fibrosis. Furthermore, this treatment dramatically suppressed TIMP-1 expression in HSCs from both model rats. These data indicate that the administration of rAAV/siRNA-TIMP-1 attenuated liver fibrosis by directly elevating the function of MMP-13 and diminishing activated HSCs. It also resulted in indirect decreased expression of type I collagen, MMP-2, and transforming growth factor-beta 1. In conclusion, rAAV/siRNA-TIMP-1 may be an effective antifibrotic gene therapy agent.