title: Adenoviral transfection of hepatocytes with the thioredoxin gene confers protection against apoptosis and necrosis.
abstract: 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.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Tsutsui
authors.forename: Hiroko
authors.initials: H
authors.lastname: Koide
authors.forename: Hiroko
authors.initials: H
authors.lastname: Fukahori
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Isoda
authors.forename: Shinji
authors.initials: S
authors.lastname: Higashiyama
authors.forename: Isamu
authors.initials: I
authors.lastname: Maeda
authors.forename: Fumi
authors.initials: F
authors.lastname: Tashiro
authors.forename: Eiji
authors.initials: E
authors.lastname: Yamato
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Miyazaki
authors.forename: Junji
authors.initials: J
authors.lastname: Yodoi
authors.forename: Masaya
authors.initials: M
authors.lastname: Kawase
authors.forename: Kiyohito
authors.initials: K
authors.lastname: Yagi
mesh_terms: 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
chemical_list: D013879:Thioredoxins