title: The Role and Mechanism of α-Klotho in the Calcification of Rat Aortic Vascular Smooth Muscle Cells.
abstract: OBJECTIVE
  To investigate the role and possible mechanism of α-Klotho in the calcification and the osteogenic transition of cultured VSMCs.
  
  
  METHODS
  VSMCs were cultured in vitro and divided into 5 groups, each using a different medium: (1) control; (2) β-GP; (3) β-GP + Klotho; (4) β-GP + LiCl; (5) β-GP + Klotho + LiCl. Calcium deposits were visualized using Alizarin Red S staining. The calcium concentrations were determined by the o-cresolphthalein complexone method. BMP2, Runx2 and β-catenin levels were estimated by western blotting, and the level of α-SMA was determined by using immunofluorescence at day 12.
  
  
  RESULTS
  β-GP induced an increase in the expression of BMP2, Runx2, and β-catenin. The calcium content increased, and the expression of α-SMA decreased. Alizarin Red S staining was positive under the high phosphorus conditions. BMP2, Runx2, and β-catenin levels and the calcium content decreased when the cells were cultured with rmKlotho; however, the levels of each were upregulated after treatment with the LiCl.
  
  
  CONCLUSIONS
  Klotho can ameliorate the calcification and osteogenic transition of VSMCs induced by β-GP. The mechanism of Klotho in preventing calcification in VSMCs may be partially mediated by the inhibition of the Wnt/β-catenin signaling pathway.
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China ; Department of Nephrology, The First People's Hospital of Changzhou, The Third Affiliated Hospital of Soochow University, Changzhou 213000, China.
authors.forename: Tianlei
authors.initials: T
authors.lastname: Chen
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Mao
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Cheng
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wu
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Ningning
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Xiufen
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Qian
authors.affiliation: Department of Nephrology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.
authors.forename: Changying
authors.initials: C
authors.lastname: Xing
mesh_terms: D000818:Animals; D001011:Aorta / Q000378:metabolism* / Q000502:physiology; D055396:Bone Morphogenetic Protein 2 / Q000378:metabolism; D002113:Calcification, Physiologic / Q000502:physiology*; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D005966:Glucuronidase / Q000378:metabolism*; D000090265:Klotho Proteins; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000378:metabolism* / Q000502:physiology*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology; D015854:Up-Regulation / Q000502:physiology; D061205:Vascular Calcification / Q000378:metabolism* / Q000503:physiopathology; D051176:beta Catenin / Q000378:metabolism
chemical_list: C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D050659:Core Binding Factor Alpha 1 Subunit; D051176:beta Catenin; D005966:Glucuronidase; D000090265:Klotho Proteins