Transplantation of human thioredoxin gene-modified hepatocytes for treatment of acute liver failure in rat model.
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 osteosarcoma 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 MTT 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.


CONCLUSIONS
hTrx gene-modified hepatocyte transplantation can effectively alleviate acute liver failure in rats.
Li
Hua
H
Liver Transplantation Center, Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China.
Jiang
Nan
N
Zhang
Jian
J
Wang
Gen-shu
GS
Yang
Yang
Y
Chen
Gui-hua
GH
D000818:Animals; D022781:Hepatocytes / Q000378:metabolism* / Q000637:transplantation*; D006801:Humans; D017114:Liver Failure, Acute / Q000628:therapy*; D051379:Mice; D041681:NIH 3T3 Cells; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013879:Thioredoxins / Q000235:genetics*
D013879:Thioredoxins