TITLE:
Long-term culture of primary porcine mature hepatocytes in the medium supplemented with ascorbic acid 2-phosphate
AUTHORS:
Yohichi Kumaki, Iku Kumaki, Xiaomei Guo, Weilin Shang, Toshie Koyama, Ai Okamura, Yoshiaki Shiba, Toshiyuki Mukaiyama, Noriko Sasaki, Makoto Kodama
KEYWORDS:
Albumin Secretion; Ammonium Metabolic Activity; Ascorbic Acid 2-Phosphate; Porcine Hepatocytes; Reverse Transcriptase-Polymerase Chain Reaction
JOURNAL NAME:
Natural Science,
Vol.2 No.11,
November
30,
2010
ABSTRACT: In this study, the effect of ascorbic acid 2-phosphate (Asc2P) was tested on porcine and rat mature hepatocytes in vitro. a). Asc2P increased the porcine, but not rat, albumin secretion and mRNA expression. The enhancing effect of Asc2P on porcine C/EBP alpha mRNA was observed in porcine mature hepatocytes. These data suggested that Asc2P played an important role in the regulation of porcine albumin mRNA level. b). The enhancing effect of Asc2P on ammonium metabolic activity was also observed in porcine, but not rat, mature hepatocytes. The porcine ornithine transcarbamylase (OTC) and arginase mRNAs were augmented by Asc2P, indicating that Asc2P had a direct effect on the urea cycle. c). The porcine collagen type I and type III mRNA, but not type XII mRNA, were detected as well, sugessting that Asc2P did not have the effect on the non-parenchymal hepatocytes to induce collagen type I and III mRNA expression. d). Our RT-PCR analysis demonstrated that the porcine hepatocytes expressed the sodium-ascorbate co-transporters SVCT1 and SVCT2, however, the intensities of porcine sodium-ascorbate co-transporters SVCT1 and SVCT2 bands were not changed markedly. These findings indicated that the Asc2P had no effect on SVCT1 and SVCT2 mRNA expression. e). The enhancing effect of Asc2P on porcine albumin mRNA was inhibited by staurosporine, a portein kinase inhibitor. We conclude that the enhanced albumin mRNA by Asc2P might be due to activation of tyrosine protein kinase and/or PKC and the Asc2P enhanced porcine albumin mRNA mainly at the transcriptional step.