Inhibition of ATIR by shRNA prevents collagen synthesis in hepatic stellate cells.
Currently, strategies aimed at disrupting renin-angiotensin-aldosterone system (RAAS) are extensively investigated for treating liver fibrosis. However, the experiment results remain unsatisfactory, mainly due to excessive level of angiotensin II (AngII) in gene expression. In this article, we aim to investigate whether suppression of AngII-type I receptor (ATIR) expression by short hairpin RNA (shRNA) expression vectors decreases the level of collagen synthesis in hepatic stellate cells (HSCs). Three pairs of ATIR-targeted shRNA expression vectors were transfected into HSC-T6 cells. Compared with the control group, both mRNA and protein levels of ATIR expression were significiently decreased in shRNA-treated groups, and the inhibitory effect exhibited a dose- and time-dependent pattern. Accordingly, TGF-β1 mRNA expression in shRNA1 group was reduced by about 54% compared with the control group. The level of Procollagen type III, hyaluronic acid, and laminin declined by about 46.4, 52.6, and 42%, respectively. In conclusion, shRNA expression vectors targeting ATIR could attenuate collagen synthesis.
Dong
Peihong
P
Department of Infectious Diseases, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou, Zhejiang, China.
Yu
Fujun
F
Fan
Xufei
X
Lin
Zhuo
Z
Chen
Yongping
Y
Li
Ji
J
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003094:Collagen / Q000096:biosynthesis*; D017931:DNA Primers; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D012313:RNA / Q000235:genetics*; D051381:Rats; D044140:Receptor, Angiotensin, Type 1 / Q000187:drug effects* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017931:DNA Primers; D044140:Receptor, Angiotensin, Type 1; D012313:RNA; D003094:Collagen