title: Acortatarin A inhibits high glucose-induced extracellular matrix production in mesangial cells.
abstract: BACKGROUND
  Diabetic nephropathy (DN) is the leading cause of end-stage renal disease. Various treatment regimens and combinations of therapies provide only partial renoprotection. Therefore new approaches are needed to retard the progression of DN. The aim of the present study was to evaluate the role of a novel spiroalkaloid from Acorus tatarinowii named acortatarin A (AcorA) in inhibiting high glucose-induced extracellular matrix accumulation in mesangial cells (MCs).
  
  
  METHODS
  The cytotoxity of AcorA on MCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide (MTT) assay. The expression of fibronectin and collagen IV was examined by real time PCR and western blotting. The expression of p22(phox) and p47(phox) was detected by western blot. The interaction between p22(phox) and p47(phox) was examined by co-immunoprecipitation. The phosphorylation of p47(phox) was examined by immunoprecipitation. The phosphorylation of protein kinase C (PKC) α, PKCβ, phospholiase C gamma (PLCγ1), and the p85 subunit of PI3K was determined by Western blotting.
  
  
  RESULTS
  AcorA significantly inhibited high glucose-induced activation of NADPH oxidase, a ROS-generating enzyme, by increasing phosphorylation of p47(phox) and enhancing interaction between p22(phox) and p47(phox). Preincubation of AcorA with MCs inhibited high glucose-induced collagen IV and fibronectin production in a dose-dependent manner. Moreover, AcorA attenuated high glucose enhanced phosphorylation of PKCα, PKCβ, PLCγ1, and the p85 subunit of PI3K.
  
  
  CONCLUSIONS
  AcorA inhibits high glucose-induced extracellular matrix production via blocking NADPH oxidase activation.
authors.affiliation: Division of Nephrology, Nanfang Hospital, Southern Medical University, Research Institute of Nephrology Guangdong Province, Key Laboratory for Organ Failure Research, Ministry of Education, Guangzhou, Guangdong 510515, China.
authors.forename: Zhi-Fang
authors.initials: ZF
authors.lastname: Zhao
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Zhou
authors.forename: Xia
authors.initials: X
authors.lastname: Chen
authors.forename: Yong-Xian
authors.initials: YX
authors.lastname: Cheng
authors.forename: Fan-Fan
authors.initials: FF
authors.lastname: Hou
authors.forename: Jing
authors.initials: J
authors.lastname: Nie
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D005109:Extracellular Matrix / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D047468:Immunoprecipitation; D050527:Mesangial Cells / Q000187:drug effects* / Q000378:metabolism*; D009025:Morpholines / Q000378:metabolism*; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D013141:Spiro Compounds / Q000378:metabolism*
chemical_list: D009025:Morpholines; D013141:Spiro Compounds; C550646:acortatarin A; D005947:Glucose