[The effect of recombinant human hepatocyte growth factor on the functions of rat hepatocytes in primary culture].
The effect of recombinant human Hepatocyte Growth Factor (rhHGF) on gluconeogenesis, urea synthesis, ATP level and total protein amount of rat hepatocytes in primary culture was investigated for 5 days in culture. According to the rhHGF concentration in media (1,5,10 ng/ml), three groups were designed. The group added Epidermal Growth Factor (EGF) was chosen as a control group. Gluconeogenesis was increased significantly on day 1 by addition of HGF, not by a concentration dependency fashion, but urea synthesis was not activated by HGF and declined on day 5. ATP levels were maintained and kept high during the culture in the EGF group. But ATP levels in the HGF groups were lower than those of the control and significantly declined on day 5. The same tendency was observed in the total amount of protein. In the EGF group, total amount of protein was maintained throughout the experimental period, but the HGF groups could not keep the amount of protein of day 1 till day 5. Phase contrast microscopic findings also showed detachment and deformity of cells on day 5 in the HGF groups. These results suggested that HGF activated gluconeogenesis of the rat hepatocytes in primary culture in early days, but it did not activate urea synthesis, and prolonged exposure of HGF caused to decline intracellular ATP levels and led to cell death in culture condition, which was speculated that the strong effect on hepatocytes by HGF exhausted cell energy. HGF was thought to be unfavorable in the respect of maintenance of primary cultured hepatocytes, but further studies are needed to confirm this speculation.
First Department of Surgery, Hokkaido University School of Medicine, Sapporo, Japan.
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Kamachi
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D005943:Gluconeogenesis / Q000187:drug effects; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D008297:Male; D011506:Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology; D014508:Urea / Q000378:metabolism
D011506:Proteins; D011994:Recombinant Proteins; D017228:Hepatocyte Growth Factor; D000255:Adenosine Triphosphate; D014508:Urea