The Bioomics Analysis Of “Yin-deficiency-heat” Syndrome And The Study Of Traditional Chinese Herb Interventionon Of “Heat” Syndrome

According to Traditional Chinese medicine,“heat” is a series of “heat syndrome” in the whole or part body,after the body being influenced by “bad” factors.According to the different causes and characteristics,“heat” is divided into “excessive heat” and “deficiency heat”,and “deficiency heat” is further subdivided into Yin-deficiency-heat and Qi-deficiency-heat.Yin-deficiency constitution is a common sub-health state of the human body.However,due to the lack of objective quantitative diagnostic indicators,patients in the Yin-deficiency constitution cannot be corrected in time and can often develop into Yin-deficiency-heat syndrome.Therefore,it is necessary to apply modern diagnostic techniques to identify the biological markers for the impoertant stages of Yin-deficiency-heat syndrome.In the present study,we performed Solexa sequencing and non-targeted metabolomics analysis using high performance liquid chromatography coupled with mass spectrometry to screen differentially expressed m RNAs and differential metabolites in the individuals in Yin-deficiency constitution,Yin-deficiency-heat syndrome,Zhibai Dihuang granule treated group and healthy control.Bioinformatics methods were used to perform enrichment analysis of differentially expressed m RNAs and differential metabolites for biological functions and signaling pathways.Furthermore,we found that the differentially expressed m RNAs and differential metabolites were related to the energy metabolism and lipid metabolism.Real-time PCR was used to validate the m RNA expression in the serum of subjects in Yin-deficiency constitution and Yindeficiency-heat syndrome.We found that the mitochondrially encoded NADH dehydrogenase 2(MT-ND2)m RNA as a potential biomarker for screening individuals with Yin-deficiencyheat syndrome from Yin-deficiency constitution(sensitivity of 85.7 % and a specificity of 60.0 %)was differentially expressed in the serum of the individuals.MT-ND2 can make the clinical use of medicine which increases Yin to extinguish heat more accurate.Receiver operating characteristic(ROC)curve and logistic regression analysis was used to evaluate the efficacy of the diagnostic model based on differential metabolites.We combined the three metabolites such as Glycine,Sphingomyelin,and Isocitrate to establish the model with a sensitivity of 85.3 % and a specificity of 80.0 % to diagnose Yin-deficiency constitution.We combined the four metabolites such as Decanoyl-L-carnitine,Dodecanoylcarnitine,phosphatidylcholine and Oligopeptide(Asp Arg Pro)to establish the model with a sensitivity of 87.8 % and a specificity of 81.8 % to diagnose Yin-deficiency-heat syndrome.After Zhibai Dihuang granule treating,glucose and Decanoyl-L-carnitine showed a tendency to return,and hypoxanthine showed differential metabolism.Our study provides potential biomarker for the diagnosis of stages of Yin-deficiency-heat syndrome and also provides a new method for the quantification and objectification of disease in traditional Chinese medicine(TCM).Andrographis paniculata(AP)has been widely used in China for centuries to treat various diseases,and especially to treat inflammation caused by excessive heat.Inflammation is the common biological characteristic of excessive heat caused by pathogen infection,mental factors and bad living habits.Diterpenoid lactones are the main anti-inflammatory components of AP.However,systematic chemical composition and biological activities,as well as key pharmacophores,of these diterpenoid lactones from AP have not yet been clearly understood.In this study,17 diterpenoid lactones were analyzed by SAR,and most of them attenuated the generation of TNF-? and IL-6 in LPS-induced RAW 264.7 cells examined by ELISA.Pharmacophores of diterpenoid lactones responsible for the anti-inflammatory activities were revealed based on the quantitative structure-activity relationship(QSAR)models.Moreover,AP-1 and AP-4 exerted anti-inflammatory activity in LPS microinjection-induced zebrafish,which might be correlated with the inhibition of the translocation of NF-?B p65 from cytoplasm to nucleus.Our study provides guidelines for future structure modification and rational drug design of diterpenoid lactones with anti-inflammatory properties in medical chemistry.