Study On Cold Or Heat Property Of Fructus Aurantii Immaturus Based On Energy Metabolism And Whole Genome Expression

The property of Chinese materia medica is a comprehensive expression of the biological effects of medicine acting on the body. The material composition and their combination of Chinese Materia Medica are the foundation of the properties, and the condition of the body is a prerequisite of the medicinal efficacy. In this study, anminal experiments were used to detect the cold or heat property of Fructus Aurantii Immaturus and we applied gene chip technology to study the basic property of Fructus Aurantii Immaturus for the first time. Thus we could explore the mechanism of pharmacologic action and lay the foundation of clinical medicationObjective: To discuss the influences on energy metabolism of asthenia cold syndrome rat models by Fructus Aurantii Immaturus, explore the objective indexes of cold or heat property and their relationships of Fructus Aurantii Immaturus. To detect the alteration of liver gene expression profiles of asthenia cold syndrome rat model interfered by Fructus Aurantii Immaturus, Rhizoma Zingiberis and Rheum Officinale. To analyse the molecular mechanism, the related therapeutic effects of Chinese materia medica with cold or heat properties, and detect the property of Fructus Aurantii Immaturus as well, thus provide a more scientific basis for the clinical medication.Methods: Asthenia cold syndrome rat models were induced by Chinese medicine compound. The indexes included appearance, temperature, weight and autonomic activities were observed and recorded during the process. After model building, the drug intervention were performed. The content of lactic acid, pyruvic acid and ATPase activity of liver tissue were determined by chemical colorimetric spectrophotometer. The content of T3, T4, TSH and TRH in blood serum were detected by enzyme-linked immunosorbent assay(ELISA).All the detection indexes were included to establish regression model and predict the cold or heat property of Fructus Aurantii Immaturus. The rat liver gene expressions in eachgroup were detected, we selected the differential expression genes, analyzed the genes by GO and KEGG pathway, and conducted the significant analysis on the genetic function of differential genes. A part of genes were selected to test the accuracy of results by RT-qPCR.Results: PLSR analysis showed that the sample points of model group and control group were completely separated, the points of Rhizoma Zingiberis group were close to control group and Rheum Officinale group were close to model group, while the points of Fructus Aurantii Immaturus group were distributed on both sides of the center. There were1745 strips of differential expression gene in asthenia cold model group compared to control group. Enrichment analysis of GO and KEGG showed that differential genes mainly involved immune response, defense response and some special metabolism processes. There were 1986 strips of differential expression gene in Fructus Aurantii Immaturus group compared to model group. Significant gene function of GO analysis showed that differential genes almost all belonged to metabolism processes. KEGG analysis showed some important singnal passway involved such as steroid hormone biosynthesis, antigen processing and presentation, Allograft rejection, etc. 867 strips of differential expressions gene were found in Rhizoma Zingiberis group, Fructus Aurantii Immaturus group compared to model group. 672 strips were found in Rheum Officinale group, Fructus Aurantii Immaturus group compared to model group. Among these differential expressions genes, the fold changes in Fructus Aurantii Immaturus group were less than Rhizoma Zingiberis group or Rheum Officinale group. There were 507 strips of coexpression differential gene among three medicated group compared to model group,and gene expression quantities in Fructus Aurantii Immaturus group were between Rhizoma Zingiberis group and Rheum Officinale group.Conclution: The main symptoms and signs of model rats were consistent with the clinical diagnosis of asthenia cold syndrome which covered the pathological features of asthenia cold syndrome. Scatter spot of PLSR showed that the intervention effects of Fructus Aurantii Immaturus fell between Rhizoma Zingiberis and Rheum Officinale, and Fructus Aurantii Immaturus could correct asthenia cold state to some extent, combinding with predicted score we concluded that Fructus Aurantii Immaturus was warm in nature.Many stimulus-response-related genes were up-down regulated in asthenia cold syndrome rats, which could induce the decline of immune response and defense response. The molecular mechanism of “insufficiency of yang qi” in asthenia cold syndrome waspossibly related to these genes. Fructus Aurantii Immaturus improved the energy metabolism process of model rats mainly through regulating the related genes of the metabolism of amino acid, protein and some steroid. The regulation of genes on immun-related biological process would have an impact on immune response and defence response. These genes were possibly related to the molecular mechanism of improving asthenia cold syndrome. The regulation of some genes in Fructus Aurantii Immaturus group fell between Rhizoma Zingiberis group and Rheum Officinale group, and had more coexpression gene in Rhizoma Zingiberis group compared to Rheum Officinale group. The results of gene analysis were consistent with PLSR analysis, that is to say, Fructus Aurantii Immaturus was warm in nature.