title: Quantitative evaluation of Oryeongsan and its action on water regulation in renal inner medullary collecting duct cells.
abstract: BACKGROUND
  Oryeongsan (ORS, Wulingsan) has been reported to possess renal protective effects from renal diseases such as diabetes-induced renal damage, and nephrocalcinosis.
  
  
  OBJECTIVE
  This study was conducted to evaluate the quantitative analysis and the inhibitory effect of ORS on hypertonic stress-induced water channel and apoptosis in murine inner medullary collecting duct cell line (mIMCD-3).
  
  
  METHODS
  Chromatographic and NMR spectroscopic analysis were performed and water balance regulation was determined by Western blot, RT-PCR, and immunofluorescnece.
  
  
  RESULTS
  Seven active principles (5-hydroxymethylfurfural, alismoxide, methyl(-)trans-cinnamate, adenine, guanosine, adenosine, and ferulic acid) in ORS were isolated and the structures were identified mainly by NMR spectroscopic analysis. In addition, contents of these metabolites in ORS were evaluated by HPLC analysis. Pretreatment with ORS significantly attenuated the hypertonic stress (175mM NaCl)-induced increase in protein levels of AQP2 and apical membrane insertion. ORS also attenuated osmolyte sodium-myo-inositol transporter (SMIT) expression and tonicity-responsive enhancer binding protein (TonEBP) mRNA under hypertonic stress. Those actions of ORS presented the similar effect of PKA inhibitor which AQP2 expression throughout the inhibition of vasopressin-mediated cAMP/PKA signal pathway. On the other hand, pretreatment with ORS attenuated hypertonic stress-induced cell death. Hypertonic stress-induced Bax or caspase-3 expression was decreased by ORS, resulting in anti-apoptotic effect.
  
  
  CONCLUSIONS
  The present data suggest that the beneficial effect of ORS in water balance and apoptosis against hypertonic stress of renal collecting ducts.
authors.affiliation: Department of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan 570-749, Republic of Korea; Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: shrons@wku.ac.kr.
authors.forename: Yun Jung
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea; KM Fundamental Research Division, Korea Institute of Oriental Medicine, 1672 Yuseong-daero, Yuseong-Gu, 34054 Daejeon, Republic of Korea. Electronic address: dasonya@hanmail.net.
authors.forename: So Min
authors.initials: SM
authors.lastname: Lee
authors.affiliation: Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea; College of Pharmacy, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: shrons@naver.com.
authors.forename: Xiang
authors.initials: X
authors.lastname: Cui
authors.affiliation: Department of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan 570-749, Republic of Korea; Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: morality16@hanmail.net.
authors.forename: Jung Joo
authors.initials: JJ
authors.lastname: Yoon
authors.affiliation: Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea; College of Pharmacy, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: hoh@wku.ac.kr.
authors.forename: Hyun Cheol
authors.initials: HC
authors.lastname: Oh
authors.affiliation: Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea; College of Pharmacy, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: yckim@wonkwang.ac.kr.
authors.forename: Youn Chul
authors.initials: YC
authors.lastname: Kim
authors.affiliation: Department of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan 570-749, Republic of Korea; Department of Oriental Medical Ophthalmology & Otolaryngology & Dermatology, College of Oriental Medicine, Wonkwang University, Shinyong-dong, Iksan, Jeonbuk 570-749, Republic of Korea. Electronic address: spinx11@wku.ac.kr.
authors.forename: Min Cheol
authors.initials: MC
authors.lastname: Park
authors.affiliation: Department of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan 570-749, Republic of Korea; Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: dgkang@wku.ac.kr.
authors.forename: Dae Gill
authors.initials: DG
authors.lastname: Kang
authors.affiliation: Department of Oriental Medicine and Professional Graduate School of Oriental Medicine, Wonkwang University, Iksan 570-749, Republic of Korea; Hanbang Body-fluid Research Center, Wonkwang University, Iksan 570-749, Republic of Korea. Electronic address: host@wku.ac.kr.
authors.forename: Ho Sub
authors.initials: HS
authors.lastname: Lee
mesh_terms: D000818:Animals; D051399:Aquaporin 2 / Q000235:genetics / Q000378:metabolism; D019921:Biotinylation; D002460:Cell Line; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000502:physiology; D007685:Kidney Tubules, Collecting / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D051379:Mice; D009997:Osmotic Pressure / Q000187:drug effects; D014867:Water / Q000378:metabolism*
chemical_list: C496024:Aqp2 protein, mouse; D051399:Aquaporin 2; D004365:Drugs, Chinese Herbal; C000614392:oryeongsan; D014867:Water
keywords: AQP2; Apoptosis; Hypertonicity; Oryeongsan; mIMCD-3