強皮症線維芽細胞を用いた線維化の機序の解明

2010 Fiscal Year Annual Research Report

• Project/Area Number: 20591350
• Research Institution: Kumamoto University
• Principal Investigator: 尹 浩信 熊本大学, 生命科学研究部, 教授 (20282634)
• Keywords: collagen / TGF-β / extracellular matrix / signal transduction / transcriptional regulation

Research Abstract

正常および強皮症培養線維芽細胞を用いて免疫プロット法にてCD36蛋白の発現量を検討したところ強皮症線維芽細胞において発現が低下していた。皮膚組織を用いた免疫染色にて強皮症線維芽細胞におけるCD36蛋白の発現を検討したところCD36発現低下がin vivoにて確認された。またreal-time quantitative PCR法にてCD36 mRNA発現の低下が認められた。CD36蛋白の発現低下はトロンボスポンジン-1の過剰発現、I型コラーゲン過剰発現、α-smooth muscle actin過剰発現、Smad3のリン酸化亢進と相関した。また正常線維芽細胞ではTGF-β刺激にてCD36発現低下が確認され、強皮症線維芽細胞におけるCD36の発現低下はautocrine TGF-β signaling(強皮症線維芽細胞自身が分泌するTGF-βが活性化してTGF-β情報伝達が亢進した結果)によるものと考えられた。また正常線維芽細胞におけるsiRNA CD36を用いたRNA干渉によるCD36発現抑制によってトロンボスポンジン-1の過剰発現、I型コラーゲン過剰発現、α-smooth muscle actin過剰発現、Smad3のリン酸化亢進が誘導され、CD36発現抑制は線維化を誘導する可能性が示唆された。強皮症線維芽細胞におけるCD36発現低下の意義をさらに明らかにするため、正常および強皮症線維芽細胞へのlatent form TGF-β添加によってCD36の発現量、TGF-β活性化、トロンボスポンジン-1の発現量、I型コラーゲン発現童、α-smooth muscle actin発現量、Smad3のリン酸化の程度を検討した。正常線維芽細胞ではlatent form TGF-β添加によってCD36の発現低下、TGF-β活性化、トロンボスポンジン-1の発現亢進、I型コラーゲン発現亢進、α-smooth muscle actin発現亢進、Smad3のリン酸化亢進が確認されたが、強皮症線維芽細胞においてはCD36の発現量、TGF-β活性化、トロンボスポンジン-1の発現量、I型コラーゲン発現量、α-smooth muscle actin発現量、Smad3のリン酸化の程度に変化を認めなかった。さらに強皮症線維芽細胞におけるCD36とトロンボスポンジン-1の介合、CD36とlatent form TGF-βの介合、トロンボスポンジン-1とlatent form TGF-βが確認されたが、正常線維芽細胞では無刺激下では各々認められなかった。

Research Products

• [Journal Article] Circulating miR-29a levels in patients with scleroderma-spectrum disorder. (2011)
  • Author(s): Kawashita Y, Jinnin M, Makino T, Kajihara I, Makino K, Honda N, Masuguchi S, Fukushima S, Inoue Y, Ihn H.
  • Journal Title: J Dermatol Sci Volume: 61 Pages: 67-69
  • Peer Reviewed
• [Journal Article] Altered dynamics of TGF-β receptors in scleroderma fibroblasts. (2011)
  • Author(s): Asano Y, Ihn H, Jinnin M, Tamaki K.
  • Journal Title: Ann Rheum Dis Volume: 70 Pages: 384-387
  • Peer Reviewed
• [Journal Article] CD109 expression levels in human malignant melanoma. (2010)
  • Author(s): Ohshima Y, Yajima I, Yanagishita T, Kumasaka MY, Yanagishita T, Watanabe D, Takahashi M, Inoue Y, Ihn H, Matsumoto Y, Kato M.
  • Journal Title: J Dermatol Sci Volume: 57 Pages: 132-146
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• [Journal Article] Prevention of ultraviolet B radiation-induced epidermal damage by expression of heat shock protein 70. (2010)
  • Author(s): Matsuda M, Hoshino T, Yamashita Y, Tanaka KI, Maji D, Sato K, Adachi H, Sobue G, Ihn H, Funasaka Y, Mizushima T.
  • Journal Title: J Biol Chem Volume: 285 Pages: 5848-5858
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• [Journal Article] The expression of human high molecular weight melanoma-associated antigen in acral lentiginous melanoma. (2010)
  • Author(s): Nishi H, Inoue Y, Kageshita T, Takata M, Ihn H.
  • Journal Title: Bioscience Trends Volume: 4 Pages: 86-89
  • Peer Reviewed
• [Journal Article] Transglutaminase 1 preferred substrate peptide K5 is an efficient tool in diagnosis of lamellar ichthyosis. (2010)
  • Author(s): Akiyama M, Sakai K, Yanagi T, Fukushima S, Ihn H, Hitomi K, Shimizu H.
  • Journal Title: Am J Pathol Volume: 176 Pages: 1592-1599
  • Peer Reviewed
• [Journal Article] Up-regulated type I collagen expression by the inhibition of Rac1 signaling pathway in human dermal fibroblasts. (2010)
  • Author(s): Igata T, Jinnin M, Makino T, Moriya C, Muchemwa F, Ishihara T, Ihn H.
  • Journal Title: Biochem Biophys Res Commun Volume: 393 Pages: 101-105
  • Peer Reviewed
• [Journal Article] Supression of melanin production by expression of HSP70. (2010)
  • Author(s): Hoshino T, Matsuda M, Yamashita Y, Takehara M, Fukuya M, Mineda K, Maji D, Ihn H, Adachi H, Sobue G, Funasaka Y, Mizushima T.
  • Journal Title: J Biol Chem Volume: 285 Pages: 13254-13263
  • Peer Reviewed
• [Journal Article] Basic fibroblast growth factor stimulates the proliferation of human dermal fibroblasts via the ERK1/2 and JNK pathways. (2010)
  • Author(s): Makino T, Jinnin M, Muchemwa F, Fukushima S, Ihn H.
  • Journal Title: Br J Dermatol Volume: 162 Pages: 717-723
  • Peer Reviewed
• [Journal Article] Serum levels of soluble vascular endothelial growth factor receptor-2 in patients with-systemic sclerosis. (2010)
  • Author(s): Jinnin M, Makino T, Kajihara I, Honda N, Makino K, Ogata A, Ihn H.
  • Journal Title: Br J Dermatol Volume: 162 Pages: 751-758
  • Peer Reviewed
• [Journal Article] A novel COL1A1 exon 14/PDGFB fusion gene in dermatofibrosarcoma protuberans (2010)
  • Author(s): Muchemwa FC, Jinnin M, Wakasugi S, Sakamoto M, Inoue Y, Ishihara T, Ihn H.
  • Journal Title: Eur J Dermatol Volume: 20 Pages: 390-391
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• [Journal Article] PDGFB quantification is a useful tool in the diagnosis of dermatofibrosarcoma protuberans : a study of 10 cases. (2010)
  • Author(s): Muchemwa FC, Wakasugi S, Honda Y, Ihn H.
  • Journal Title: Clip Exp Dermatol Volume: 35 Pages: 295-299
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• [Journal Article] Inflixmab monotherapy in Japanese patients with moderate-to-severe plaque psoriasis and psoriatic arthritis.A randomized, double-blind, placebo-controlled multicenter trial. (2010)
  • Author(s): Torii H, Nakagawa H, Iizuka H, Aoyagi S, Okuyama R, Ohtsuki M, Abe M, Takamori K, Matsuba S, Terui T, Igarashi A, Kawashima M, Amagai M, Takae Y, Eto H, Ikeda S, Komine M, Saeki H, Hayakawa K, Kitami A, Watanabe H, Takehara K, Shimade S, Kitajima Y, Takigawa M, Morita A, Yamanaka K, Takahashi K, Tarutani M, Tarsi M, Kubota.Y, Nakayama J, Ihn H.
  • Journal Title: J Dermatol Sci Volume: 59 Pages: 40-49
  • Peer Reviewed
• [Journal Article] Amelanoti malignant melanoma where primary lesion was discovered 5 ears after metastasis in the lymph node. (2010)
  • Author(s): Kato T, Inoue Y, Ishihara T, Kageshita T, Ihn H.
  • Journal Title: J Dermatol Volume: 37 Pages: 730-734
  • Peer Reviewed
• [Journal Article] Characterization of monocyte/macrophage subsets in the skin and peripheral blood derived from patients with systemic sclerosis. (2010)
  • Author(s): Higashi-Kuwata N, Jinnin M, Makino T, Fukushima S, Inoue Y, Muchemwa FC, Yonemura Y, Komohara Y, Takeya M, Mitsuya H, Ihn H.
  • Journal Title: Arhritis Res Ther Volume: 12 Pages: R128(1-10)
  • Peer Reviewed
• [Journal Article] Serum TARC/CCL17 levels are increased in dermatomyositis associated with interstitial lung disease. (2010)
  • Author(s): Kawashima T, Tada Y, Asano Y, Yazawa N, Tomita M, Tamaki Z, Kubo M, Ihn H, Sugaya M, Kadono T, Tamaki K, Sato S.
  • Journal Title: J Dermatol Sci Volume: 60 Pages: 52-54
  • Peer Reviewed
• [Journal Article] Impaired lymphangiogenesis due to excess VEGF-D/Flt-4 signaling in the skin of patients with systemic sclerosis. (2010)
  • Author(s): Honda N, Jinnin M, Kajihara I, Makino T, Fukushima T, Ihn H.
  • Journal Title: Br J Dermatol Volume: 163 Pages: 776-780
  • Peer Reviewed
• [Journal Article] Down-regulation of mir-424 contributes to the abnormal angiogenesis via MEK1 and cyclin E1 in senile hemangioma : its implications to therapy. (2010)
  • Author(s): Nakashima T, Jinnin M, Etoh T, Fukushima S, Masuguchi S, Maruo K, Inoue Y, Ishihara T, Ihn H.
  • Journal Title: PloS One Volume: 5 Pages: e14334
  • Peer Reviewed