title: Metabolic profiling reveals therapeutic biomarkers of processed Aconitum carmichaeli Debx in treating hydrocortisone induced kidney-yang deficiency syndrome rats.
abstract: BACKGROUND
  Kidney-yang deficiency syndrome (KYDS) is a diagnostic pattern in traditional Chinese medicine (TCM) and clinical data showed that the unbalance in adrenal cortical hormone is the key issue in KYDS patients. The processed Ranunculaceae aconitum carmichaeli debx (bai-fu-pian in Chinese, BFP) is one of the most commonly used Chinese herbs for treating KYDS. The present study was conducted to explore the therapeutic biomarkers of the BFP in treating hydrocortisone administration induced KYDS rats.
  
  
  METHODS
  Thirty male Sprague-Dawley rats were randomly divided into five groups with six in each group. KYDS in rats was induced by i.p. injection of hydrocortisone at the dose of 10mg/kg per day for 15 days as described previously. The rats with KYDS were administered orally, starting from the day of hydrocortisone administration stopped, with BFP extract at the dose of 0.32g/kg, 0.64g/kg and 1.28g/kg per day respectively for 15 days. The blood samples were collected for the liquid chromatography quadruple time-of-flight mass spectrometry (LC-Q-TOF-MS) test, as well as radioimmunoassay to determine the concentrations of cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP) and adrenocorticotrophic hormone (ACTH). The metabolic responses to BFP administration were investigated by using the principal components analysis (PCA) and orthogonal partial least squares analysis (OPLS). Bioinformatics analyses were performed by using the Ingenuity Pathway Analysis (IPA). Variance analysis and linear regression analysis were used in this study.
  
  
  RESULTS
  The signs and concentrations of cAMP, cGMP and ACTH in the model rats were similar to those previously described about KYDS rats and BFP treatment can reverse the changes. Seventeen significantly changed metabolites among different groups were identified. Thirteen metabolites were identified in the KYDS rats comparing to healthy rats with nine up-regulated and four down-regulated. After BFP treatment at three dosages, five up-regulated metabolites including phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide were dose-dependently reversed. The network analysis with IPA showed that four canonical pathways including superpathway of methionine degradation, purine nucleotides de novo biosynthesis II, tyrosine synthesis and serotonin receptor signaling involved the therapeutic mechanism of BFP in treating the KYDS rats.
  
  
  CONCLUSIONS
  Five therapeutic biomarkers (phosphate, betaine, (4-hydroxyphenyl) acetaldehyde, 5-hydroxyindol-3-acetic acid and 5'-phosphoribosyl-N-formylglycinamide) and two corresponding canonical pathways (amino acid metabolism and purine nucleotide metabolism) were identified to be involved in the therapeutic mechanism of BFP treating the KYDS.
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Tan
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai 200433, China.
authors.forename: Xinru
authors.initials: X
authors.lastname: Liu
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China; School of Chinese Medicine, Hong Kong Baptist University, 7 Baptist University Road, Kowloon Tong, Hong Kong.
authors.forename: Cheng
authors.initials: C
authors.lastname: Lu
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: He
authors.affiliation: Beijing University of Chinese Medicine, Beijing 100700, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: China-Japan Friendship Hospital, Beijing 100030, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Xiao
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Miao
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China; Hunan University of Chinese Medicine, Changsha 410208, China.
authors.forename: Ke
authors.initials: K
authors.lastname: Zhou
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai 200433, China.
authors.forename: Zhongxiao
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, Second Military Medical University, Shanghai 200433, China. Electronic address: wdzhangy@hotmail.com.
authors.forename: Weidong
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Institute of Basic Research in Clinical Medicine, China Academy of Chinese Medical Sciences, Beijing 100700, China; School of Chinese Medicine, Hong Kong Baptist University, 7 Baptist University Road, Kowloon Tong, Hong Kong. Electronic address: lap64067611@126.com.
authors.forename: Aiping
authors.initials: A
authors.lastname: Lu
mesh_terms: D031841:Aconitum / Q000737:chemistry*; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D019295:Computational Biology; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D006854:Hydrocortisone / Q000633:toxicity; D007674:Kidney Diseases / Q000188:drug therapy* / Q000503:physiopathology; D016014:Linear Models; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008516:Medicine, Chinese Traditional; D010936:Plant Extracts / Q000008:administration & dosage / Q000494:pharmacology*; D025341:Principal Component Analysis; D011685:Purine Nucleotides / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013577:Syndrome; D016711:Yang Deficiency / Q000188:drug therapy* / Q000503:physiopathology
chemical_list: D000596:Amino Acids; D015415:Biomarkers; D010936:Plant Extracts; D011685:Purine Nucleotides; D006854:Hydrocortisone
keywords: Kidney-Yang deficiency syndrome rats; Metabolic profiling; Processed Aconitum carmichaeli debx; Therapeutic biomarkers