2003 Fiscal Year Annual Research Report

酸化ストレスによる情報伝達変化と神経細胞傷害の分子構構

Research Project

Project/Area Number	14570201
Research Institution	Tokai University
Principal Investigator	竹腰進東海大学, 医学部, 講師 (70216878)
Co-Investigator(Kenkyū-buntansha)	長村義之東海大学, 医学部, 教授 (10100992) 滝澤俊也東海大学, 医学部, 助教授 (70197234)
Keywords	diacylglycerol / protein kinase C / lipid peroxidation / signal tansduction / oxidative stress / MAP kinase / BIAcore / glutathione peroxidase
Research Abstract	プロテインキナ-ゼC(PKC)は、イノシトール燐脂質情報伝達系のkey enzymeであり、種々のlipid mediatorの中でもジアシルグリセロール(DAG)によって活性化されることにより様々な細胞応答に重要な役割を果たしている。我々はDAGが過酸化されることにより強力な活性化物質に変質することを証明した。本研究では、過酸化DAGが特定のアイソザイムの活性化を介して病変を引き起こすことを、アデノウイルスベクターを用いたPKCαおよびδ高発現細胞を用いて検討した。その結果、過酸化DAGは、PKCδを過剰発現させた初代培養神経細胞においてMAPキナーゼカスケードの異常活性化を誘導し神経細胞傷害を引き起こすことが明らかとなった。また、過酸化水素、t-BHPなどの単なる過酸化物は傷害作用を示さず、脂溶性情報伝達物質の酸化物が特に強い傷害作用を有することが判明した。一方、PKCαあるいはPKCδkinase negative(kinase活性を持たないmutant δ分子)を高発現した初代培養神経細胞では、過酸化DAGによる神経細胞傷害は全く認められなかった。これらの結果から、過酸化DAGはPKCδ過剰活性化→MAP kinaseカスケード異常活性化を介して神経細胞傷害を引き起こすと考えられた。また、BIAcore装置を用いた検討により、過酸化DAGの強いPKC活性化作用はPKC分子と高い親和性を有するためであることが明らかとなった。更に、グルタチオンペルオキシダーゼ活性を有する薬剤であるebselenは過酸化DAGによる傷害作用を有意に抑制した。また、セレン添加および欠乏条件下で過酸化DAGの作用を検討したところ、セレン欠乏条件下でのみ神経細胞傷害が誘導された。

Research Products
(16 results)

All Other

All Publications (16 results)

[Publications] Robert Yoshiyuki Osamura, et al.: "Mechanisms for Production and Secretion of Hormones in Physiologic and Pathologic Conditions"Acta Histochemca Cytochemica. 36. 99-103 (2003)
[Publications] Yasutomo Sekido, et al.: "Heterogeneous gene alterations in primary breast cancer contribute to discordance between primary and asynchronous metastatic/recurrent sites : HER2 gene amplification and p53 mutation."International Journal of Oncology. 22. 1225-1232 (2003)
[Publications] Tsutomu Kanehira, et al.: "Kinobeon A as a Potent Tyrosinase Inhibitor from Cell Culture of Safflower : In vitro Comparisons of Kinobeon A with Other Putative Inhibitors."Planta Medica. 69. 457-459 (2003)
[Publications] Johbu Itoh, et al.: "Simultaneous Detection of TUNEL and Methyl Green Signals on Apoptotic Nuclei in Rat Intestine Absorptive Epithelial Cells by Confocal Laser Scanning Microscopy."Acta Histochemca Cytochemica. 36. 367-376 (2003)
[Publications] R.Yoshiyuki.Osamura, et al.: "Endocrine Pathoology in Translational Medicine : An Overview of Current and Future Prospects."Endocrine Pathology. 14. 117-121 (2003)
[Publications] Akira Matsuno, et al.: "Dynamics of Subcellular Organelles, Growth Hormone, Rab3b, SNAP-25, and Syntaxin in Rat Pituitary Cells Caused by Growth Hormone Releasing Hormone and Somatostatin."Microscopy Research and Technique. 62. 232-239 (2003)
[Publications] Susumu Takekoshi, et al.: "Lipid Peroxidation Involvement in the Alteration of Protein Kinase C Signaling"Acta Histochemca Cytochemica. 36. 281-286 (2003)
[Publications] Shinobu Umemura, et al.: "Methyl green Staining and DNA Strands in vitro : High Affinity of Methyl Green Dye to Cytosine and Guanine."Acta Histochem Cytochem. 36. 361-366 (2003)
[Publications] Tsutomu Kanehira, et al.: "A Novel and Potent Biological Antioxidant, Kinobeon A, from Cell Culture of Safflower"Life Sciences. 74. 87-97 (2003)
[Publications] R.Yoshiyuki.Osamura, et al.: "Application of gene targeting technologies for the study of pituitary development and neoplasms"Acta Histochemca Cytochemica. 36. 249-254 (2003)
[Publications] Ichiro Mori, et al.: "Exogenous calcitonin suppresses growth fraction of thyroid C cells"Acta Histochemca Cytochemica. 36. 439-442 (2003)
[Publications] Akira Matsuno, et al.: "Production of pituitary hormone by human pituitary adenoma is under autocrine and paracrine regulationof hypothalamic hormones secreted from adenoma cells"Acta Histochemca Cytochemica. 36. 415-420 (2003)
[Publications] Takashi Hirasawa, et al.: "Chemotherapeutic Effects on p53 Immunoexpression in Ovarian Cancer : Difference between Two Chemotherapy Regimens"Acta Histochemca Cytochemica. in press. (2004)
[Publications] Hidetaka Nagata, et al.: "Quercetin Enhances Melanogenesis by Increasing Activity and Synthesis of Tyrosinase in Human Melanoma Cells"Pigment Cell Research. in press. (2004)
[Publications] Reiko Takeyama, et al.: "Quercetin induced melanogenesis in a reconstituted three-dimensional human epidermal model"Journal of Molecular Histology. in press. (2004)
[Publications] 竹腰進(分担): "組織化学2003 組織化学シグナル増強法どうすれば強い反応を得ることができるか"日本組織細胞化学会. 11 (2003)

2003 Fiscal Year Annual Research Report

酸化ストレスによる情報伝達変化と神経細胞傷害の分子構構

Principal Investigator

竹腰 進 東海大学, 医学部, 講師 (70216878)

Research Products

[Publications] Robert Yoshiyuki Osamura, et al.: "Mechanisms for Production and Secretion of Hormones in Physiologic and Pathologic Conditions"Acta Histochemca Cytochemica. 36. 99-103 (2003)

[Publications] Yasutomo Sekido, et al.: "Heterogeneous gene alterations in primary breast cancer contribute to discordance between primary and asynchronous metastatic/recurrent sites : HER2 gene amplification and p53 mutation."International Journal of Oncology. 22. 1225-1232 (2003)

[Publications] Tsutomu Kanehira, et al.: "Kinobeon A as a Potent Tyrosinase Inhibitor from Cell Culture of Safflower : In vitro Comparisons of Kinobeon A with Other Putative Inhibitors."Planta Medica. 69. 457-459 (2003)

[Publications] Johbu Itoh, et al.: "Simultaneous Detection of TUNEL and Methyl Green Signals on Apoptotic Nuclei in Rat Intestine Absorptive Epithelial Cells by Confocal Laser Scanning Microscopy."Acta Histochemca Cytochemica. 36. 367-376 (2003)

[Publications] R.Yoshiyuki.Osamura, et al.: "Endocrine Pathoology in Translational Medicine : An Overview of Current and Future Prospects."Endocrine Pathology. 14. 117-121 (2003)

[Publications] Akira Matsuno, et al.: "Dynamics of Subcellular Organelles, Growth Hormone, Rab3b, SNAP-25, and Syntaxin in Rat Pituitary Cells Caused by Growth Hormone Releasing Hormone and Somatostatin."Microscopy Research and Technique. 62. 232-239 (2003)

[Publications] Susumu Takekoshi, et al.: "Lipid Peroxidation Involvement in the Alteration of Protein Kinase C Signaling"Acta Histochemca Cytochemica. 36. 281-286 (2003)

[Publications] Shinobu Umemura, et al.: "Methyl green Staining and DNA Strands in vitro : High Affinity of Methyl Green Dye to Cytosine and Guanine."Acta Histochem Cytochem. 36. 361-366 (2003)

[Publications] Tsutomu Kanehira, et al.: "A Novel and Potent Biological Antioxidant, Kinobeon A, from Cell Culture of Safflower"Life Sciences. 74. 87-97 (2003)

[Publications] R.Yoshiyuki.Osamura, et al.: "Application of gene targeting technologies for the study of pituitary development and neoplasms"Acta Histochemca Cytochemica. 36. 249-254 (2003)

[Publications] Ichiro Mori, et al.: "Exogenous calcitonin suppresses growth fraction of thyroid C cells"Acta Histochemca Cytochemica. 36. 439-442 (2003)

[Publications] Akira Matsuno, et al.: "Production of pituitary hormone by human pituitary adenoma is under autocrine and paracrine regulationof hypothalamic hormones secreted from adenoma cells"Acta Histochemca Cytochemica. 36. 415-420 (2003)

[Publications] Takashi Hirasawa, et al.: "Chemotherapeutic Effects on p53 Immunoexpression in Ovarian Cancer : Difference between Two Chemotherapy Regimens"Acta Histochemca Cytochemica. in press. (2004)

[Publications] Hidetaka Nagata, et al.: "Quercetin Enhances Melanogenesis by Increasing Activity and Synthesis of Tyrosinase in Human Melanoma Cells"Pigment Cell Research. in press. (2004)

[Publications] Reiko Takeyama, et al.: "Quercetin induced melanogenesis in a reconstituted three-dimensional human epidermal model"Journal of Molecular Histology. in press. (2004)

[Publications] 竹腰進(分担): "組織化学2003 組織化学シグナル増強法 どうすれば強い反応を得ることができるか"日本組織細胞化学会. 11 (2003)

竹腰進東海大学, 医学部, 講師 (70216878)

[Publications] 竹腰進(分担): "組織化学2003 組織化学シグナル増強法どうすれば強い反応を得ることができるか"日本組織細胞化学会. 11 (2003)