Adenoviral transfection of hepatocytes with the thioredoxin gene confers protection against apoptosis and necrosis.
A recombinant adenovirus vector containing the human thioredoxin (TRX) gene was constructed using the Cre-loxP recombination system and used to transfect rat hepatocytes with very high efficiency. The TRX gene was expressed in a dose-dependent manner and significantly modulated rat cellular functions. The TRX gene conferred resistance to oxidative stress, such as hydrogen peroxide treatment, on the host hepatocytes. FACS analysis of DNA fragmentation showed that the TRX gene suppressed hepatocyte apoptosis. It also significantly extended the life span of hepatocytes cultured conventionally on polystyrene plates. Liver-specific functions were maintained in the viability-modulated hepatocytes. Moreover, TRX expression did not affect hepatocyte spheroid formation and it extensively suppressed necrosis in the internal cells. Thus, the transfection of hepatocytes with the TRX gene successfully confers global maintenance of liver functions. These findings provide important information for the development of bioartificial liver support systems and gene therapy for liver diseases.
Tsutsui
Toshio
T
Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan.
Koide
Hiroko
H
Fukahori
Hiroko
H
Isoda
Katsuhiro
K
Higashiyama
Shinji
S
Maeda
Isamu
I
Tashiro
Fumi
F
Yamato
Eiji
E
Miyazaki
Jun-Ichi
J
Yodoi
Junji
J
Kawase
Masaya
M
Yagi
Kiyohito
K
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis; D002478:Cells, Cultured; D019610:Cytoprotection; D005822:Genetic Vectors; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D019164:Liver, Artificial; D008297:Male; D009336:Necrosis; D018384:Oxidative Stress; D051381:Rats; D017207:Rats, Sprague-Dawley; D018874:Spheroids, Cellular; D013879:Thioredoxins / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection
D013879:Thioredoxins