Several Tongue Proteomics Preliminary Study

Objective: Glossocopy is an important diagnosis basis of traditional Chinese medicine, and the research of tongue coating is an important content of it. The mechanism and objective of tongue coating is one of the key problems to the modernizational study of TCM. Since the completion of Human Genome Project, researches on the essence of beings and disease comes into post genome era. Functional genomics and proteomics are the most important front field of modern life sciences. Based on the fruits of our several years long research and used the technic of proteomics, this study plans to explore the mechanism of tongue coatings, and tongue coating based on the molecular holistic level. The purpose of this study is to investigate the relationship between proteomics of tongue coating, and symptoms/sign of TCM, and also to explore the molecular biological events and symptons/signs of tongue coating.Methods: Objects of observation were divided into four groups namely normal white and thin coating tongue, pathological thin fur, thick fur and peeling fur. Scraping tongue coating with spatula,then centrifugalizing and washing. Immobilized pH gradient to isoelectric focusing for the first vertical SDS-PAGE for the second to the 2DE technology links with the key factors, such as sample handling, sample volume parameters electrophoresis gel concentration SDS gel electrophoresis and staining methods for the study and optimization of the experimental conditions. Its four groups proteins of tongue coatingwere separated, silver stain color gel scanning into computers, ImageMaster 2D Platinum software analysis and comparison electrophoresis patterns. Choice of 36 different point cut, glue, after trypsin digestion, mass spectrometry (MALDI-TOF-MS) ImageMaster 2D Platinum software analysis of 2D gel images obtained. Results: This study established and optimized the 2-DE methods,and the computer assisted image analysis system was also established. Satisfactory 2-DE maps including resolution and reproducibility were obtained. These were used to compare the proteomic difference the four groups tongue coating, On average,1082±105,1052±85,1129±98and 1143±140 protein spots were obtained in map of difference the four groups tongue coating.36 peptide mass fingerprints (PMF) map was obtained by MALDI-TOF-MS. The typic peptide mass was searched in the NCBI proteins database, The two proteins undertook situ enzymolysis, we harvested successfully two finger printings via peptides finger printing analysis, the data base searching show that the two proteins were cytokeratin 13,GAL7 protein and Cystatin B.Our results of comparative proteomics analysis might provide clues for further elucidating the role of these proteins in pathological Tongue coating appearance and development potential associated biomarkers.Summary: The tongue coating proteomics research can be for clinical syndrome differentiation of TCM, curativeeffect evaluate of TCM, and study on the pharmacodynamics and drug screening of traditional Chinese medicines provide the micro indicatrix and scientific basis.