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eCAM Advance Access originally published online on June 15, 2007
eCAM 2008 5(4):463-474; doi:10.1093/ecam/nem049
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© 2007 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

A Proteomic Approach for the Diagnosis of ‘Oketsu’ (blood stasis), a Pathophysiologic Concept of Japanese Traditional (Kampo) Medicine

Chinami Matsumoto1, Tetsuko Kojima1, Kazuo Ogawa1, Satoshi Kamegai2, Takuya Oyama2, Yukari Shibagaki2, Tetsuo Kawasaki2, Hiroshi Fujinaga3, Kozo Takahashi3, Hiroaki Hikiami6, Hirozo Goto6, Chizuru Kiga4,5, Keiichi Koizumi5, Hiroaki Sakurai5,7, Hiroshi Muramoto2, Yutaka Shimada6,7, Masahiro Yamamoto1, Katsutoshi Terasawa8, Shuichi Takeda1 and Ikuo Saiki5,7

1Central Research Laboratories, Tsumura & Co., Ibaraki, 2Bioinformatics Division, INTEC Web and Genome Informatics Corporation, Toyama, 3Department of Japanese Oriental Medicine, Toyama Prefectural Central Hospital, Toyama, 4Toyama New Industry Organization, Toyama, 5Division of Pathogenic Biochemistry, Institute of Natural Medicine, 6Department of Kampo Medicine, Faculty of Medicine and 7The 21st Century COE Program, University of Toyama, Toyama and 8Department of Japanese-Oriental (Kampo) Medicine, Graduate School of Medicine, Chiba University, Chiba, Japan

‘Oketsu’ is a pathophysiologic concept in Japanese traditional (Kampo) medicine, primarily denoting blood stasis/stagnant syndrome. Here we have explored plasma protein biomarkers and/or diagnostic algorithms for ‘Oketsu’. Sixteen rheumatoid arthritis (RA) patients were treated with keishibukuryogan (KBG), a representative Kampo medicine for improving ‘Oketsu’. Plasma samples were diagnosed as either having an ‘Oketsu’ (n = 19) or ‘non-Oketsu’ (n = 29) state according to Terasawa's ‘Oketsu’ scoring system. Protein profiles were obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) and hierarchical clustering and decision tree analyses were performed. KBG treatment for 4 or 12 weeks decreased the ‘Oketsu’ scores significantly. SELDI protein profiles gave 266 protein peaks, whose expression was significantly different between the ‘Oketsu’ and ‘non-Oketsu’ states. Hierarchical clustering gave three major clusters (I, II, III). The majority (68.4%) of ‘Oketsu’ samples were clustered into one cluster as the principal component of cluster I. The remaining ‘Oketsu’ profiles constituted a minor component of cluster II and were all derived from patients cured of the ‘Oketsu’ state at 12 weeks. Construction of the decision tree addressed the possibility of developing a diagnostic algorithm for ‘Oketsu’. A reduction in measurement/pre-processing conditions (from 55 to 16) gave a similar outcome in the clustering and decision tree analyses. The present study suggests that the pathophysiologic concept of Kampo medicine ‘Oketsu’ has a physical basis in terms of the profile of blood proteins. It may be possible to establish a set of objective criteria for diagnosing ‘Oketsu’ using a combination of proteomic and bioinformatics-based classification methods.

Keywords: proteinchip – surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) – keishibukuryogan (KBG)

For reprints and all correspondence: Ikuo Saiki, Division of Pathogenic Biochemistry, University of Toyama, 2630 Sugitani, Toyama 930-0194, Japan. Tel: +81-76-434-7620; Fax: +81-76-434-5058; E-mail: byosei{at}inm.u-toyama.ac.jp

Received May 19, 2006; accepted April 13, 2007


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