Transplantation of human thioredoxin gene-modified hepatocytes for treatment of acute liver failure in rat model
10.3760/cma.j.issn.0366-6999.2009.21.020
- Author:
Hua LI
1
;
Nan JIANG
;
Jian ZHANG
;
Gen-Shu WANG
;
Yang YANG
;
Gui-Hua CHEN
Author Information
1. Liver Transplantation Center,Third Affiliated Hospital of Sun Yat-sen University,Guangzhou,Guangdong 510630,China
- Keywords:
hepatocyte;
transplantation;
thioredoxin;
gene expression;
liver failure;
acute
- From:
Chinese Medical Journal
2009;(21):2631-2635
- CountryChina
- Language:Chinese
-
Abstract:
Background Mostly because of the limited number and proliferative ability of the transplanted hepatocytes,hepatocyte transplantation offers only temporary support to the hepatic function with rather poor functional replacement of the damaged liver parenchyma.This study aimed to observe the therapeutic effect of human thioredoxin(hTrx)gene-modified hepatocytes on experimental acute liver failure in rats.Methods hTrx cDNA was obtained by reverse transcription-polymerase chain reaction(RT-PCR)from human osteosercoma 143(TK-)cells to construct the recombinant retrovirus vector pLEGFP/hTrx,which was packaged into PA317 cells to collect the recombinant retrovirus containing hTrx gene.After titration and characterization,the recombinant retrovirus was applied to primary cultured rat hepatocyte for infection to generate hTrx gene-modified rat hepatocytes,whose viability and antioxidative capacity were examined by immunohistochemistry and MIF assay,respectively.In a Sprague-Dawley(SD)rat model of acute liver failure,the modified hepatocytes were injected into the spleen,and the hepatic function and survival rate of the recipient rats were evaluated at different time points after the transplantation.Results NIH3T3 cells infected by the recombinant retrovirus were capable of expressing bioactive hTrx in the form of fusion proteins.Immunohistochemistry demonstrated normal function of the hTrx gene-modified hepatocytes,which possessed strong antioxidative capacity as shown by MTT assay.Transplantation of the modified hepatocytes in rats with acute liver failure resulted in significantly lowered serum alanine aminotransferase(ALT)and total bilirubin(TBIL)levels(P＜0.05).The hepatocytes exhibited long-term survival and efficient proliferation after transplantation.Fourteen days after the operation,the rat models receiving hTrx gene-modified hepatocytes had significantly higher survival rate than those without the transplantation.Conclusion hTrx gene-modifled hepatocyte transplantation can effectively alleviate acute liver failure in rats.
