Analyzing expressed genes of HEK cells cultured with “hot nature” Chinese herbs by gene chips

• Main Authors: You Hong-Yi, 游恆懿
• Other Authors: Chen Guan-Wei
• Format: Others
• Language: zh-TW
• Published: 2002
• Online Access: http://ndltd.ncl.edu.tw/handle/84091522157035274338

碩士 === 中國醫藥學院 === 中西醫結合研究所 === 90 === Traditional Chinese medicine ( TCM ) has been extensively used to treat many diseases in eastern society throughout thousands of years. However, the diagnostic and therapeutic concepts were not identical to those of modern western medicine. The major principles of TCM are treating patients based on the clinical disease patterns. For example, cold disease patterns could be treated with so-called hot medicines and they have the ability to relieve internal coldness of the human body, increase metabolic rate, sympathetic tone, etc. In the past few years, a new technology, DNA microarray ( DNA chip ), has made it possible to simultaneously study the expression of thousands genes in a single chip. In this pilot study, we try to elucidate the gene expression of the HEK 293 cell in vitro under the treatment by 2 traditional Chinese medicine ( Rdix Acconiti and Zingiberis Rhizoma ), which belongs to the “hot” medicine. For this, gene expression of herbs treated cells and none treated cells was compared simultaneously using Dr. Peck Konan microarray system. Dual color staining method of enzyme colorimetry detection system was utilized by biotin-16-dUTP and dig-11-dUTP during reverse transcription. The dual color immuno-chemical detection followed the antibody/enzyme/substrate pairs; that is, Streptavidin conjugated beta-galactosidase with X-gal substrate and ant-dig with alkaline phosphates with Fast Red TR/Naphthol AS-MX substrate. A color microarray image was rendered by the chromogens yielded by the enzymatic reactions. The result showed changes in expressions of 47 genes up regulation and 46 genes down regulation by Radix Acconiti; 105 genes up regulation and 31 genes down regulation by Zingiberis Rhizoma.. This study established a method to the understanding of the genetic influence of TCM. Further studies are needed to verify these results.