Targeted inhibition of platelet-derived growth factor receptor-beta subunit in hepatic stellate cells ameliorates hepatic fibrosis in rats.
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.
Chen
S-W
SW
Department of Gastroenterology, Changzheng Hospital, Second Military Medical University, Shanghai, China.
Chen
Y-X
YX
Zhang
X-R
XR
Qian
H
H
Chen
W-Z
WZ
Xie
W-F
WF
D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D005355:Fibrosis; D020868:Gene Silencing; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D055166:Hepatic Stellate Cells / Q000378:metabolism* / Q000473:pathology; D008297:Male; D023421:Models, Animal; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000379:methods
D005904:Glial Fibrillary Acidic Protein; D034741:RNA, Small Interfering; D020797:Receptor, Platelet-Derived Growth Factor beta