Targeted inhibition of platelet-derived growth factor receptor-β subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats
Targeted inhibition of platelet-derived growth factor receptor-β subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats
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DOI：
10.1038/gt.2008.93
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发表时间：
2008-11-01
期刊：
影响因子：
5.1
通讯作者：
Xie, W-F
中科院分区：
文献类型：
--
作者：
Chen, S-W;Chen, Y-X;Xie, W-F
肝星状细胞（HSCs）的活化是肝纤维化发病的关键事件。血小板衍生生长因子（PDGF）是造血干细胞最有效的丝裂原，PDGF受体- β亚基（pdgfr - β）是PDGF诱导造血干细胞增殖所必需的。本研究合成了一种高基因沉默效能的pdgfr - β小干扰RNA (siRNA)，在体外可抑制pdgfr - β的表达，抑制其活化和增殖，但不能诱导造血干细胞凋亡。为了避免pdgfr - β非特异性干扰的副作用，我们构建了一种hscs特异性短发夹RNA （shRNA）表达质粒，其中pdgfr - β shRNA由胶质纤维酸性蛋白（GFAP）启动子驱动。双染色免疫荧光检测表明GFAP启动子可靶向四氯化碳诱导的急性损伤大鼠肝脏和胆管结扎（BDL）诱导的慢性损伤大鼠肝脏的hsc转基因表达。此外，造血干细胞特异性pdgfr - β shRNA可减轻BDL所致大鼠模型的肝损伤和肝纤维化。本研究表明pdgfr - β siRNA可能是一种抗纤维化药物。GFAP启动子诱导hscs特异性RNA干扰的应用可能为肝纤维化的细胞特异性基因治疗提供新的有力工具。基因治疗（2008）15,1424-1435；doi: 10.1038 / gt.2008.93;2008年5月29日在线发布
The activation of hepatic stellate cells (HSCs) is the key event of the pathogenesis of hepatic fibrosis. Platelet-derived growth factor (PDGF) is the most potent mitogen for HSCs, and PDGF receptor-beta subunit (PDGFR-beta) is required for the proliferation of HSCs induced by PDGF. In this study, a high gene-silencing-efficacy PDGFR-beta small interference RNA (siRNA) was synthesized that could suppress the PDGFR-beta expression and inhibit the activation and proliferation but could not induce the apoptosis of HSCs in vitro. To avoid the side effect of nonspecific interference of PDGFR-beta, we constructed an HSCs-specific short hairpin RNA (shRNA) expression plasmid in which PDGFR-beta shRNA was driven by a glial fibrillary acidic protein (GFAP) promoter. The double-staining immunofluorescence examination indicated that GFAP promoter could target the transgene expression into HSCs in carbon tetrachloride induced acute injured rat's liver and bile duct ligation (BDL)-induced chronic injured rat's liver. Furthermore, HSCs-specific PDGFR-beta shRNA could relieve liver injury and hepatic fibrosis in the rat's model induced by BDL. This study demonstrates that PDGFR-beta siRNA may be presented as an antifibrogenic agent. The application of HSCs-specific RNA interference induced by the GFAP promoter might supply a new powerful tool for cell-specific gene therapy of hepatic fibrogenesis. Gene Therapy (2008) 15, 1424-1435; doi:10.1038/gt.2008.93; published online 29 May 2008