1989 Fiscal Year Annual Research Report

培養肝細胞を用いた機能と増殖を調節する肝組織構築の生化学的研究

Research Project

Project/Area Number	61440032
Research Institution	University of Tokushima
Principal Investigator	市原明徳島大学, 酵素センター, 教授 (40035374)
Co-Investigator(Kenkyū-buntansha)	富田優美子徳島大学, 酵素センター, 助手 (00089913) 田中啓二徳島大学, 酵素センター, 助手 (10108871) 野田千征子徳島大学, 酵素センター, 助教授 (40035506)
Keywords	肝細胞 / 増殖因子 / 初代培養 / 細胞接触 / Transfoming Growth Factor(TGF-beta) / Interleukin(IL-1) / cyclic AMP / ホルボ-ルエステル
Research Abstract	肝再生機構の根本として初代培養肝細胞を用いその増殖因子を分離同定しその構造もcDNAから明らかとなった。その分子量やサブユニット構造は蛋白構造と変わらなかったが、アミノ酸配列には既知蛋白との相同は無く新らしい増殖因子と考えられる。従来この因子はラット血小板から放出されると考えられたが、そのcDNAによるノ-ザン法、増殖促進活性でラット各種組織を検討したが、肝臓のみならず肺、脾臓など多様な組織に分布していた。また再生肝や毒物による障害肝で肝非実質細胞に増加する。この事は肝再生時に非実質細胞に増殖因子が増加、放出され実質細胞の増殖を促進するパラクリン機構が考えられる。次にこの増殖促進には低細胞密度で培養する必要があり、高細胞密度ではc-mycの発現は抑えられ、また分化機能のセリン脱水酵素のmRNAの発現も高かった。一方増殖抑制因子の作用はTGF-betaでは低細胞密度(増殖相)で、IL-1は高細胞密度(分化相)で抑制効果が強かった。これら増殖の細胞内シグナル伝達機構としてAキナ-セやCキナ-セの関与が考えられるがcAMP添加は低細胞密度で増殖を促進し、ホルボ-ルエステル(TPA)は高細胞密度で促進した。この様に肝細胞の増殖は細胞接触で強く調節されており、肝再生機構として成熟肝臓組織では実質細胞は周囲の細胞やマトリックスと密に接触し分化機能を発現し静止している(Go相)。しかし肝障害がおこるとこの接触がゆるみG相に移り増殖因子の効果を受ける様になりS期に入る。この時期には分化機能は抑制されていて、こうして増殖が進むと接触が回復し、更に増殖抑制因子の効果も出て細胞はGoにもどり分化し再生が完成する。この様に肝再生調節にその組織構築の重要性が明らかとなり更にその細胞内機構にAやCキナ-セの関与が示唆された。

Research Products

(16 results)

All Other

All Publications (16 results)

[Publications] Inoue,C.: "Nicotinamide prolongs survival of primary cultured hepatocytes without involving loss of hepatocyte-specific functions" Journal of Biological Chemistry. 264. 4747-4750 (1989)
[Publications] Okada,F.: "Purification and strucutre analysis of a latent form of transforming growth factor-beta from rat platelets" Journal of Biochemistry. 106. 304-310 (1989)
[Publications] Ichihara,A.: "Regulation of protein and amino acid metabolism in cultured hepatocytes" Nutrion:Protein and Amino Acids(Eds.Naito,H.et al.)(Japanese Science Society Press,Tokyo). 49-65 (1990)
[Publications] Ichihara,A.: "Hormonal regulation of gene expressions in primary cultured hepatocytes and its relation with cell growth" Proceedings of Conference on Molecular Biology and Gene Engeneerings. 4. 16-20 (1989)
[Publications] Okada,F.: "One of two subunits of masking protein in latent TGF-beta is a part of pro-TGF-beta" FEBS Letter. 242. 240-244 (1989)
[Publications] Tanaka,K.: "Direct evidence for nuclear and cytoplasmic co-localization of proteasomes(Multi-protease complex)in liver." Journal of Cellular Physiology. 139. 34-41 (1989)
[Publications] Tanaka,K.: "Autodegradation of rat liver proteasomes(Large multi-catalytic proteinase complex)" Biochemical Biophysical Research Communication. Vol.158.548-554 (1989)
[Publications] Matthews,W.: "Involvement of proteasome in different degradative processes in mammalian cells" Proceedings of National Academy of Sciences,USA. Vol.86. 2597-2601 (1989)
[Publications] Tanaka,K.: "Half-life of proteasomes(Multi-protease complexes)in rat liver" Biochemical and Biophysical Research Communication. 159. 1309-1315 (1989)
[Publications] Seol,J.H.: "Na^+,K^+-specific inhibition of protein and peptide hydrolyses by proteasomes from human hepatoma tissues" FEBS Letter. 247. 197-200 (1989)
[Publications] Fujiwara,T.: "Molecular cloning of cDNA for proteasomes(multicatalytic proteinase complex)from rat liver:Primary strucutre of the largest component(C2)" Biochemistry. 28. 7332-7340 (1989)
[Publications] Noda,C.: "Developmental and growth-related regulation of expression of serine dehydratase mRNA in rat liver" Biochemical and Biophysical Research Communication. (1990)
[Publications] Arakaki,R.: "Reciprocal, cell density dependent inhibitions of growth of rat hepatocytes in primary cultures by transforming growth factor-beta and interleukin-1beta" Experimental Cell Research. (1990)
[Publications] Noda,C.: "Structure and organization of the rat serine dehydratase gene:Possible production of two mRNAs from the single gene" Journal of Biological Chemistry. (1990)
[Publications] Andoh,T.: "Relation between serume hepatocyte growth factor and regeneration of liver after treatment with carbon tetrachloride" Hepatology. (1990)
[Publications] Kanda,S.: "Negative regulation of autonomous growth of neonatal rat hepatocytes in primary culture by interleukins and beta-type transforming growth factor" Cancer Research. (1990)

1989 Fiscal Year Annual Research Report

培養肝細胞を用いた機能と増殖を調節する肝組織構築の生化学的研究

Principal Investigator

市原 明 徳島大学, 酵素センター, 教授 (40035374)

Research Products

[Publications] Inoue,C.: "Nicotinamide prolongs survival of primary cultured hepatocytes without involving loss of hepatocyte-specific functions" Journal of Biological Chemistry. 264. 4747-4750 (1989)

[Publications] Okada,F.: "Purification and strucutre analysis of a latent form of transforming growth factor-beta from rat platelets" Journal of Biochemistry. 106. 304-310 (1989)

[Publications] Ichihara,A.: "Regulation of protein and amino acid metabolism in cultured hepatocytes" Nutrion:Protein and Amino Acids(Eds.Naito,H.et al.)(Japanese Science Society Press,Tokyo). 49-65 (1990)

[Publications] Ichihara,A.: "Hormonal regulation of gene expressions in primary cultured hepatocytes and its relation with cell growth" Proceedings of Conference on Molecular Biology and Gene Engeneerings. 4. 16-20 (1989)

[Publications] Okada,F.: "One of two subunits of masking protein in latent TGF-beta is a part of pro-TGF-beta" FEBS Letter. 242. 240-244 (1989)

[Publications] Tanaka,K.: "Direct evidence for nuclear and cytoplasmic co-localization of proteasomes(Multi-protease complex)in liver." Journal of Cellular Physiology. 139. 34-41 (1989)

[Publications] Tanaka,K.: "Autodegradation of rat liver proteasomes(Large multi-catalytic proteinase complex)" Biochemical Biophysical Research Communication. Vol.158.548-554 (1989)

[Publications] Matthews,W.: "Involvement of proteasome in different degradative processes in mammalian cells" Proceedings of National Academy of Sciences,USA. Vol.86. 2597-2601 (1989)

[Publications] Tanaka,K.: "Half-life of proteasomes(Multi-protease complexes)in rat liver" Biochemical and Biophysical Research Communication. 159. 1309-1315 (1989)

[Publications] Seol,J.H.: "Na^+,K^+-specific inhibition of protein and peptide hydrolyses by proteasomes from human hepatoma tissues" FEBS Letter. 247. 197-200 (1989)

[Publications] Fujiwara,T.: "Molecular cloning of cDNA for proteasomes(multicatalytic proteinase complex)from rat liver:Primary strucutre of the largest component(C2)" Biochemistry. 28. 7332-7340 (1989)

[Publications] Noda,C.: "Developmental and growth-related regulation of expression of serine dehydratase mRNA in rat liver" Biochemical and Biophysical Research Communication. (1990)

[Publications] Arakaki,R.: "Reciprocal, cell density dependent inhibitions of growth of rat hepatocytes in primary cultures by transforming growth factor-beta and interleukin-1beta" Experimental Cell Research. (1990)

[Publications] Noda,C.: "Structure and organization of the rat serine dehydratase gene:Possible production of two mRNAs from the single gene" Journal of Biological Chemistry. (1990)

[Publications] Andoh,T.: "Relation between serume hepatocyte growth factor and regeneration of liver after treatment with carbon tetrachloride" Hepatology. (1990)

[Publications] Kanda,S.: "Negative regulation of autonomous growth of neonatal rat hepatocytes in primary culture by interleukins and beta-type transforming growth factor" Cancer Research. (1990)

市原明徳島大学, 酵素センター, 教授 (40035374)