The System Biology Of Cold Pattern And Heat Pattern Of Rheumatoid Arthritis In Traditional Chinese Medicine

1 ObjectiveThe research is aimed to explore the distinct molecular signatures in the rheumatoid arthritis patients with traditional Chinese medicine cold pattern and heat pattern.2 MethodsFor data analysis:This study uses data from a previous multicenter randomized controlled clinical trial. Patients were from 9 clinical centers. They were randomly divided into Westren medicine treated group and Traditional Chinese Medicine treated group. ACR20 response at 12 weeks and 24 weeks was used for evaluation of efficacy. Multivariate statistical analysis was used to investigate the relationship between patern differentiation and clinical efficacy.For transcriptomics:Twenty patients with typical TCM cold pattern and heat pattern were included in the study. Microarray technology was used to reveal gene expression profiles in CD4+T cells from the patients. The signal intensity of each expressed gene was globally normalized using the R statistics program. The ratio of cold pattern to heat pattern in RA patients at more or less than 1:2 was taken as the differential gene expression criteria. Protein-protein interaction information for these genes from databases was searched, and the highly-connected regions were detected by IPCA algorithm. The significant pathways were extracted from these subnetworks by Biological Network Gene Ontology tool.For metabolomics:The blood samples of 21 RA patients and 16 healthy volunteers were used. Blood plasma was prepared by centrifuging, then every sample was transferred to an autosampler for analysis. Partial least squares discriminant analysis (PLS-DA) was employed to find potential biomarkers differentiating the RA patients from the control participants. The processed data list was UV-scaled, mean centered and exported for chemometrics analysis using SIMCA-P. To test the statistical significance the resulting PLS-DA models were cross-validated and permutation testing was performed. Error rates were calculated using MatLab. All the data collected have been statistically analyzed with the SPSS software, the count data with Chi-square test and the measuring data with T-test.3 ResultsTranscriptomics:Twenty nine genes differentially regulated between cold pattern and heat pattern were found. Among them,7 genes were expressed significantly more in cold pattern. Biological network of protein-protein interaction information for these significant genes were searched and visualized using Cytoscape, and four highly-connected regions were detected by IPCA algorithm to infer significant complexes or pathways in the biological network. Particularly the cold pattern was related to Toll-like receptor signaling pathway. The following related pathways in heat pattern were included:Calcium signaling pathway; Cell adhesion molecules; PPAR signaling pathway, Fatty acid metabolism.Metabolomics:A multivariate data-analysis procedure resulted in the discovery of several potential biomarkers related to RA patients with cold pattern and heat pattern. The PLS-DA score plot shows that the RA patients with cold pattern and heat pattern could be divided into two groups based on their metabolite profiles. T-tests were performed on both sets of variables (P<0.05). Differences between the metabolite profiles of cold pattern and heat pattern revealed 7 potential biomarkers, which 3-Oxy propanoic acid, L-Proline, Urea,5-Oxo-proline, Ribitol, Inositol and L-Leucine were included.4 ConclusionsRA patients with cold pattern and heat pattern have distinct molecular signatures with different biological processes participating. Molecular differences between the RA Cold and RA Heat groups were found which suggest differences in apoptotic activity and protein metabolism. Both the gene and metabolite profiles have elucidated relationships between several of the markers, revealed insight in the mechanism of RA Pattern differentiation. These results suggest that better knowledge of the main biological processes involved at a given pattern in TCM might help to choose the most appropriate treatment.