Metabonomic Studies Of The Essence Of Hypothyroidism With Spleen And Kidney-yang Deficiency Syndrome And Therapeutical Effects Of Sini Decoction

Hypothyroidism is a typical disease because of the dysfunction of endocrinehormone according to the western medicine theory. The main pathogenesis is thyroidhormone deficiency or inadequate action of thyroid hormone in target tissues.According to the theory of Traditional Chinese Medicine (TCM), hypothyroidismbelongs to the scope of “consumptive disease” or “asthenic disease”, and the essentialpathogenesis of hypothyroidism is “Spleen and Kidney-Yang Deficiency Syndrome”.The main treatment for hypothyroidism at present is thyroid hormone replacementtherapy. However, due to difficulties in controlling the level of thyroid hormonesthrough use of drugs or an exogenous source of thyroid hormones, this treatmentstrategy usually leads to unexpectable side effects, which deserve our close attention.Therefore, it is still urgent to discover new modes of treatment for hypothyroidism.Sini decoction (SND) is a well-knownn formula which has certain therapeuticaleffects on hypothyroidism with “Spleen and Kidney-Yang Deficiency Syndrome”. Itconsists of Acontium carmichaeli, Glycyrrhiza uralensis and Zingiber officinale.Although SND is considered as efficient agents against hypothyroidism with “Spleenand Kidney-Yang Deficiency Syndrome”, its holistic effects assessment andmechanistic understanding are still lacking due to its complex components.The core of metabolomics is studying the holistic effects of exogenous materialson the integrated organ. The systemic thinking of metabonomics and its aim atgrasping integral function make it pillars of potential bridge between TCM andwestern medicine, which may beneficially provide an opportunity to open up theblack box between evidence-based Chinese medicine and the organism. Therefore, ametabonomic method based on UHPLC-Q-TOFMS was developed to study themetabolic changes in hypothyroid rats and the intervention effect of TCM SND. Theaim of my study is to investigate the essence of hypothyroidism with “Spleen andKidney-Yang Deficiency Syndrome” and the effect mechanism of TCM SND.1. A serum metabonomic profiling method based on ultra-performance liquidchromatography/time-of-flight mass spectrometry (UHPLC/TOF-MS) was applied toinvestigate the metabolic changes in hypothyroid rats induced by propylthiouracil(PTU) and the therapeutical effects of SND. With Significance Analysis of Microarray(SAM) for classification and selection of biomarkers,13potential biomarkers in ratserum were screened out. Furthermore, Ingenuity Pathway Analysis (IPA) was introduced to deeply analyze unique pathways of hypothyroidism that were primarilyinvolved in sphingolipid metabolism, fatty acid transportation, phospholipidmetabolism and phenylalanine metabolism. Our results demonstrated that themetabonomic approach integrating with IPA was a promising tool for providing anovel methodological cue to systemically dissect the underlying molecularmechanism of hypothyroidism with “Spleen and Kidney-Yang Deficiency Syndrome”.2. A urinary metabonomic method based on ultra performance liquidchromatography coupled to mass spectrometry was employed to explore globalmetabolic characters of hypothyroidism. Three typical hypothyroidism models(methimazole-, propylthiouracil-and thyroidectomy-induced hypothyroidism) wereapplied to elucidate the molecular mechanism of hypothyroidism.17,21,19potentialbiomarkers were identified with these three hypothyroidism models respectively,primarily involved in energy metabolism, amino acid metabolism, sphingolipidmetabolism and purine metabolism. In order to avert the interference of druginteraction between the antithyroid drugs and SND, the thyroidectomy-inducedhypothyroidism model was further used to systematically assess the therapeuticefficacy of SND on hypothyroidism. A time-dependent recovery tendency wasobserved in SND-treated group from the beginning of model to the end of treatment,suggesting that SND exerted a recovery effect on hypothyroidism in a time-dependentmanner through partially regulating the perturbed metabolic pathways.3. Based on the results from serum and urine metabonomics studies, we found thatthere is a close relationship between hypothyroidism with “Spleen and Kidney-YangDeficiency Syndrome” and lipid metabolism. Therefore, we introduced anUHPLC-TOFMS based lipidomics method to analyze the global lipid profiles ofhypothyroidism with “Spleen and Kidney-Yang Deficiency Syndrome” in ratcerebellum. Using unsupervised analysis and multivariate statistical analysis, we canseparate the Sham and Hypothyroid groups clearly and screen out23potential lipidbiomarkers related to hypothyroidism with “Spleen and Kidney-Yang DeficiencySyndrome”, primarily involved in sphingolipid metabolism, glycerophospholipidmetabolism and β-oxidation of fatty acid. And then we conducted bioinformaticsanalysis to built and simulate the lipid network of hypothyroidism with “Spleen andKidney-Yang Deficiency Syndrome”, which would be useful for the elucidation thepathological mechanism of hypothyroidism with “Spleen and Kidney-YangDeficiency Syndrome”. Based on the selected23lipid biomarkers, we systematically evaluate the therapeutical effects of SND and positive drug-T4. The results showedthat both SND and T4can convert the pathological status of hypothyroidism with“Spleen and Kidney-Yang Deficiency Syndrome” to some extent through differentpathways.In summary, our study has investigated and explained the essence ofhypothyroidism with “Spleen and Kidney-Yang Deficiency Syndrome” and the effectmechanism of TCM SND by metabolomics method and biochemical assessed values,which shed a new light to the treatment strategy on hypothyroidism with “Spleen andKidney-Yang Deficiency Syndrome”.