Introduction: The goal of this project was to develop a new approach in the use of Chinese medicine for promoting recovery from overtraining syndrome. Overtraining syndrome is a condition of fatigue and declining performance that occurs following limit-reaching training for competitive events such as the Olympics. The primary factor leading to overtraining involves failure of adaptation to physical and mental stress. Therapeutic potential of Chinese medicine for overtraining has been suggested; however, biological evidence that this holistic approach corrects the functional disorder at a molecular level is lacking. Genomic technology is a powerful tool used for revealing the molecular complexity of the response to stress and the effects of Chinese medicine treatment because it provides comprehensive information on thousands of genes that underlie the responses rather than focussing on a single gene. In the current study, we first designed a Chinese herbal formula according to the Chinese medicine diagnosis of the athletes who developed symptoms of overtraining, followed by an in vitro validation using a hypoxiaischemia cultured cell model that mimics the stress of overtraining. Finally, we examined the effects of the herbal formula on gene expression profile using DNA microarray for the purposes of identifying the coordinated responses of functional groups of genes. Methods Determination of the Chinese herbal formula: Chinese athletes who developed typical overtraining syndrome were diagnosed by Chinese medicine doctors and a herbal formula containing multiple herbs aimed at promoting the recovery was prescribed according to the principle of Chinese medicine. In vitro study using hypoxia-ischemia cell model: Human astrocytes were cultured in the present or absent of Chinese herbal formula (1 mg/ml) extract (1:100). Hypoxia-ischemia model was induced by addition of Rotenone (10 ìM) and Dithionite (1 mM) to cause deprivation of cellular ATP and oxygen. After one hour incubation, the initial cell culture condition was restored to allow the regeneration of ATP and oxygen. Apoptotic cells resulted from the challenge were detected using the TUNEL assay (Intergen, USA), and visualized with fluorescent microscopy. DNA microarray measurement: Gene expression profiles of cultured astrocytes were measured using human Gene Chip (Affymetrix, California, USA) that contains a total of 44,000 genes. After hybridization the gene chips were automatically washed and stained with streptavidin-phycoerythrin by using a fluidics system. The chips were scanned with a Hewlett-Packard GeneArray Scanner (Hewlett- Packard, California, USA). The up-regulated or down-regulated genes are determined by statistic cut off (>1.5-fold change; ANOVA with Benjam ini-Hochberg correction, P < 0.05). Results This study demonstrated the effects of Chinese medicine`s approach in promoting the recovery of overtraining at cellular and genomic levels. In the cultured cell model, the herbal formula that was designed based on the Chinese medicine diagnosis exhibited protective effects against Rotenone (10 ìM) and Dithionite (1 mM) induced apoptotic cell death (P<0.05). DNA microarray further revealed a large alteration of the gene expression profile upon the hypoxia-ischemia insult, with 256 genes upregulated and 138 down-regulated. These genes are closely associated with the pathological response to stress (oxidative stress and heat shock), inflammation, protein metabolism and ion transport. Pretreatment with the Chinese herbal formula extract significantly moderated the genomic response, suggesting the Chinese medicine approach could improve the ability of adaptation to stress. Discussion/Conclusions Application of Chinese medicine has great potential in sport medicine. In the present study, genomic technology was applied to examine the change of gene expression profile, rather than a single gene, upon Chinese medicine treatment for overtraining syndrome. Our data indicated that the Chinese herbal formula developed could moderate the excess genomic alteration and prevent apoptotic cell death induced by the insult of hypoxia-ischemia stress. The effectiveness on athletes is currently under evaluation.
© Copyright 2008 2008 International Convention on Science, Education and Medicine in Sport: Proceedings, Vol. I. Veröffentlicht von People´s Sports Publishing House. Alle Rechte vorbehalten.