Establishment of diagnostic system on Kampo Medicine with gene analysis.

2005 Fiscal Year Final Research Report Summary

• Project/Area Number: 16390160
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied pharmacology
• Research Institution: The Kitasato Institute, Oriental Medicine Research Center
• Principal Investigator: HANAWA Toshihiko The Kitasato Institute, Oriental Medicine Research Center, Professor, 東洋医学研究所, 所長 (40164892)
• Co-Investigator(Kenkyū-buntansha): SAKURAI Masatomo The Kitasato Institute, Oriental Medicine Research Center, research worker, 東洋医学研究所, 研究員 (00390732) ; HAYASAKI Tomoyuki The Kitasato Institute, Oriental Medicine Research Center, research worker, 東洋医学研究所, 研究員 (70353472) ; TAKAHASHI Yuko The Kitasato Institute, Oriental Medicine Research Center, research worker, 東洋医学研究所, 研究員 (90306572) ; MURAKAMI Kazuo Foundation for Advancement of International Science, Bio-Laboratory, Director, バイオ研究所, 所長 (70110517)
• Project Period (FY): 2004 – 2005
• Keywords: Kambo Medicine / gene expression / stratification / DNA chip / FDD / Kososan / Hangekobokuto / diagnosis

Research Abstract

Fluorescent differential display and DNA chip methods were used to identify genes whose expression changed after treatment with Kososan or Hangekobokuto. First, analysis was performed by fluorescent differential display. RNA (nucleic acids functioning as a messenger of genetic information) prepared from subjects were used as a template to synthesize cDNA by reverse transcription reaction with a fluorescent dye-labeled oligo (dT) primer. The anchor primer and arbitrary sequence primers were used for a polymerase chain reaction to obtain amplified gene products (in fluorescent differential display, polymerase chain reactions occurred as many times as the number of primers). The amplified gene products were subjected to electrophoresis to screen genes whose expression changed before and after Kampo medication, followed by gene isolation, cloning, and sequencing for identification of the genes of interest. Next, a DNA chip method was performed. For each Kampo medicine, measurement of the change in expression of these genes was validated, and specific gene expression patterns in responders to Kampo medication were identified. Genes whose expression specifically changed with each Kampo medication were classified into groups to identify groups of genes related to the pharmacological effects of Kampo medicines.
Some genes were found to be characteristically expressed in responders to each of these two Kampo medicines. A group of genes with expression profiles characteristic of responders to each of the two Kampo medicines showed no change in their expression after medication, suggesting influences of expression profiles and sequences of the genes. Particularly in responders to Kososan, some genes showed expression profiles strongly suggestive of an association with efficacy. These results suggest that gene expression profiling before medication may identify patients who are likely to benefit from Kampo medication.