title: Recombinant adeno-associated virus-mediated transfer of shRNA against Notch3 ameliorates hepatic fibrosis in rats.
abstract: Liver fibrosis, a wound healing process following all kinds of liver injuries, is characterized by excessive deposition of extracellular matrix (ECM). Our previous study revealed that Notch3 might participate in liver fibrogenesis by regulating the activation of hepatic stellate cells (HSCs). The aim of this study was to assess the effects of Notch3 shRNA on hepatic fibrosis in a rat model induced by carbon tetrachloride (CCl4) and to clarify the mechanisms underlying those effects. Recombinant adeno-associated virus type 1 (rAAV1) vector carrying Notch3 shRNA (rAAV1-Notch3-shRNA) was generated and transferred to rat livers via the tail vein. The expression of Notch3, Jagged1, Hes1 and α-SMA were detected by real-time RT-PCR and immunofluorescence. The effects of rAAV1-Notch3-shRNA on fibrosis was investigated by pathological and immunohistochemical examination. Our findings showed that Notch3, Jagged1, Hes1 and α-SMA were downregulated. This downregulation was accompanied by improved hepatic fibrosis after the inhibition of Notch3 in vivo. rAAV1-Notch3-shRNA treatment reversed the epithelial-mesenchymal transition (EMT) in fibrotic livers by decreasing the expression of transforming growth factor β1 (TGF-β1) and vimentin in a line with the increased expression of E-cadherin. The inhibition of Notch3 was not found to play a role in hepatocyte proliferation. Rather, it inhibited hepatocyte apoptosis in vivo to some extent. The results of the present study suggest that the inhibition of Notch3 can protect hepatocytes from undergoing apoptosis and attenuate liver fibrogenesis. This may be a viable therapeutic option for hepatic fibrosis.
authors.affiliation: Department of Ultrasonography, Union Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan, 430022, China.
authors.forename: Shao-Ping
authors.initials: SP
authors.lastname: Zheng
authors.forename: Yi-Xiong
authors.initials: YX
authors.lastname: Chen
authors.forename: Jun-Li
authors.initials: JL
authors.lastname: Guo
authors.forename: Dan
authors.initials: D
authors.lastname: Qi
authors.forename: Shao-Jiang
authors.initials: SJ
authors.lastname: Zheng
authors.forename: Shu-Ling
authors.initials: SL
authors.lastname: Zhang
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Weng
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D005109:Extracellular Matrix / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000378:metabolism* / Q000473:pathology; D008103:Liver Cirrhosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008106:Liver Cirrhosis, Experimental / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D008297:Male; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D015820:Cadherins; C498556:Notch3 protein, rat; D034741:RNA, Small Interfering; D000071656:Receptor, Notch3; D051880:Receptors, Notch; D053773:Transforming Growth Factor beta1
keywords: Hepatic fibrosis; epithelial-mesenchymal transition; notch3; recombinant adeno-associated virus