Adeno-associated virus mediated interferon-gamma inhibits the progression of hepatic fibrosis in vitro and in vivo.
OBJECTIVE
To investigate the effects of adeno-associated virus (AAV) mediated expression of human interferon-gamma for gene therapy in experimental hepatic fibrosis in vitro and in vivo.


METHODS
We constructed the recombinant AAV encoding human INF-gamma (rAAV- INF-gamma) and took the primary rat hepatic stellate cells and carbon tetrachloride induced rats as the experimental hepatic fibrosis model in vitro and in vivo. Immunocytochemistry analysis was used to reveal the expression of alpha-SMA, the marker protein expressed in hepatic stellate cells. The mRNA expression of TGF-beta, TIMP-1, and MMP-13 were analyzed by RT-PCR method. In vivo study, the hydroxyproline content in liver and serum AST, ALT were also detected.


RESULTS
In vitro study, AAV vector could mediated efficient expression of human INF-gamma, which inhibit the activation of hepatic stellate cells, decrease the expression of alpha-SMA and mRNA of TIMP-1, TGF-beta, with the MMP-13 unchanged. In vivo study, the histological examination revealed that rAAV- INF-gamma could inhibit the progression of the hepatic fibrosis. In the rAAV-INF-gamma induced group, the hydroxyproline content and serum AST, ALT level were decreased to 177+/-28 microg/g wet liver, 668.5+/-140.0, 458.4+/-123.5 U/L, compare with the fibrosis control group 236+/-31 microg/g wet liver, 1 019.1+/-276.3, 770.5+/-154.3 U/L, respectively (P<0.01). mRNA expression of TIMP-1 in the rAAV-INF-gamma induced rat liver was decreased while no significant change was observed in TGF-beta and MMP-13.


CONCLUSIONS
All these results indicated that rAAV-INF-gamma has potential effects for gene therapy of hepatic fibrosis, which could inhibit the progression of hepatic fibrosis.
Jiangsu Key Lab of Drug Metabolism and Pharmacokinetics, China Pharmaceutical University, No. 24 Tongjiaxiang, Nanjing 210009, Jiangsu Province, China.
Miao
M
Chen
Guang-Ji
GJ
Wang
Yong
Y
Diao
Rui-An
RA
Xu
Hai-Tang
HT
Xie
Xin-Yan
XY
Li
Jian-Guo
JG
Sun
D000818:Animals; D002252:Carbon Tetrachloride Poisoning / Q000517:prevention & control*; D002478:Cells, Cultured; D017364:Collagenases / Q000235:genetics; D000229:Dependovirus / Q000235:genetics / Q000276:immunology; D004195:Disease Models, Animal; D018450:Disease Progression; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology*; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D008106:Liver Cirrhosis, Experimental / Q000517:prevention & control*; D008111:Liver Function Tests; D053509:Matrix Metalloproteinase 13; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics; D016212:Transforming Growth Factor beta / Q000235:genetics; D014771:Virion / Q000276:immunology
D012333:RNA, Messenger; D011994:Recombinant Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D016212:Transforming Growth Factor beta; D007371:Interferon-gamma; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; C507083:Mmp13 protein, rat