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Effect of Saikokeishito, a Kampo medicine, on hydrogen peroxide-induced premature senescence of normal human dermal fibroblasts

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Journal of Integrative Medicine 2014年第6期12卷 495-503页

作者： Takanobu Takata Yoshiharu Motoo naohisa tomosugi Medical Research Institute of Kanazawa Medical University Department of Medical Oncology Kanazawa Medical University Department of Nephrology Kanazawa Medical University

OBJECTIVE： Saikokeishito（TJ-10） is a Kampo（traditional Japanese herbal） medicine, clinically used for hundreds of years in East Asia. Among its various mechanisms elucidated so far, TJ-10 inhibits the production of transforming growth factor-β1（TGF-β1） and development of pancreatic fi brosis in vivo. Oxidative damage of normal human dermal fi broblasts（NHDFs） in the corium is a cause of human dermal senescence. Our aim was to determine whether TJ-10 protects NHDFs from premature senescence by hydrogen peroxide（H2O2）. METHODS： Premature senescence was induced in NHDFs by 200 μmol/L H2O2 for 4 h. Cell viability and the expressions of p53, AMP-activated protein kinase α1（AMPKα1）, AMPKα2, and 14-3-3 protein sigma（14-3-3 σ） were measured in NHDFs treated with TJ-10 for 48 h before exposure to H2O2 for 4 h. RESULTS： Cell viability after treatment with 200 μmol/L H2O2 for 4 h was similar（about 80%） to after pre-treatment with TJ-10. Ascorbic acid as a control did not protect NHDFs from damage by 200 μmol/L H2O2. Treatment with 200 μmol/L H2O2 tended to up-regulate p53 and to down-regulate SIRT1 and AMPKα1, but had no effect on AMPKα2 and 14-3-3 σ expression. Pretreatment with TJ-10 inhibited H2O2-induced up-regulation of p53 and enhanced AMPKα1 expression. CONCLUSION： It is suggested that Saikokeishito has a protective effect on oxidative stressinduced senescence of NHDFs.

关键词： Saikokeishito medicine Kampo premature senescence p53 fibroblasts

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Analysis of hepcidin expression: In situ hybridization and quantitative polymerase chain reaction from paraffin sections

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World Journal of Gastroenterology 2012年第28期18卷 3727-3731页

作者： Yuhki Sakuraoka Tokihiko Sawada Takayuki Shiraki Kyunghwa Park Yuhichiro Sakurai naohisa tomosugi Keiichi Kubota Second Department of Surgery School of Medicine Dokkyo Medical University Kitakobayashi 880 Mibu Shimotsuga Tochigi 321-0293 Japan Proteomics Research Unit Division of Advanced Medicine Medical Research Institute Kanazawa Medical College Daigaku 1-1 Uchinada Kanazawa 920-0293 Japan

AIM: To establish methods for quantitative polymerase chain reaction (PCR) for hepcidin using RNAs isolated from paraffin-embedded sections and in situ hybridization of hepatocellular carcinoma (HCC). METHODS: Total RNA from paraffin-embedded sections was isolated from 68 paraffin-embedded samples of HCC. Samples came from 54 male and 14 female patients with a mean age of 66.8 ± 7.8 years. Quantitative PCR was performed. Immunohistochemistry and in situ hybridization for hepcidin were also performed. RESULTS: Quantitative PCR for hepcidin using RNAs isolated from paraffin-embedded sections of HCC was performed successfully. The expression level of hepcidin mRNA in cancer tissues was significantly higher than that in non-cancer tissues. A method of in situ hybridization for hepcidin was established successfully, and this demonstrated that hepcidin mRNA was expressed in non-cancerous tissue but absent in cancerous tissue. CONCLUSION: We have established novel methods for quantitative PCR for hepcidin using RNAs isolated from paraffin-embedded sections and in situ hybridization of HCC.

关键词： Hepcidin Expression In situ hybridization Immunohistochemistry Real-time polymerase chain reaction

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