title: Oxymatrine inhibits aldosterone-induced rat cardiac fibroblast proliferation and differentiation by attenuating smad-2,-3 and-4 expression: an in vitro study.
abstract: BACKGROUND
  We previously demonstrated oxymatrine, an alkaloid from the Chinese medicine radix Sophorae flavescentis, ameliorates hemodynamic disturbances and cardiac fibrosis; however, the underlying mechanisms are unclear. Here, we investigated the effect and mechanism of action of oxymatrine on aldosterone-induced cardiac fibroblast to myofibroblast differentiation in vitro.
  
  
  METHODS
  Cardiac fibroblasts were isolated purified from neonatal Sprague Dawley rats. The optimal concentration of aldosterone to stimulate cardiac fibroblast proliferation was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cardiac fibroblasts were pretreated with 7.57 × 10(-4) mol/L or 3.78 × 10(-4) mol/L oxymatrine or without oxymatrine for 2 h, and then coincubated with 1 × 10(-8) mol/L aldosterone for 48 h. The MTT assay and Masson staining were used to detect the cardiac fibroblast proliferation and myofibroblast differentiation. The secretion of type I and III collagen was measured by commercial ELISA kits, and the hydroxyproline content was determined by the colorimetric assay. Western blotting assayed the Smad-2, Smad-3, and Smad-4 protein expression in cardiac fibroblasts.
  
  
  RESULTS
  The present results confirmed that aldosterone induced cardiac fibroblast to myofibroblast proliferation and differentiation. The MTT assay and Masson staining indicated oxymatrine significantly inhibited aldosterone-induced cardiac fibroblast proliferation and myofibroblast differentiation. Oxymatrine significantly inhibited aldosterone-induced secretion of type I and III collagen, as indicated by commercial ELISA kits, and aldosterone-induced increase in hydroxyproline content, as indicated by a colorimetric assay. Western blotting revealed oxymatrine attenuated aldosterone-induced Smad-2, Smad-3, and Smad-4 expression in cardiac fibroblasts.
  
  
  CONCLUSIONS
  Oxymatrine can inhibit cardiac fibroblast proliferation and differentiation into myofibroblasts via a mechanism linked to attenuation of the Smad signaling pathway.
authors.affiliation: Department of Pharmacology of Materia Medica, Guizhou Medical University, Huaxi University town, Guian New District, Guizhou, 550025, China.|The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Lingyun
authors.initials: L
authors.lastname: Fu
authors.affiliation: The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Yini
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Pharmacology of Materia Medica, Guizhou Medical University, Huaxi University town, Guian New District, Guizhou, 550025, China.|The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Tu
authors.affiliation: Department of Pharmacology of Materia Medica, Guizhou Medical University, Huaxi University town, Guian New District, Guizhou, 550025, China.|The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Haifeng
authors.initials: H
authors.lastname: Huang
authors.affiliation: The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Pharmacology of Materia Medica, Guizhou Medical University, Huaxi University town, Guian New District, Guizhou, 550025, China.|The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Tao
authors.affiliation: Department of Pharmacology of Materia Medica, Guizhou Medical University, Huaxi University town, Guian New District, Guizhou, 550025, China. shenxiangchun@126.com.|The Key Laboratory of Optimal Utilization of Natural Medicinal Resources, Guizhou Medical University, Huaxi University town, Guian new district, Guizhou, 550025, China. shenxiangchun@126.com.
authors.forename: Xiangchun
authors.initials: X
authors.lastname: Shen
mesh_terms: D000450:Aldosterone / Q000494:pharmacology*; D000470:Alkaloids / Q000737:chemistry / Q000494:pharmacology*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000187:drug effects*; D009206:Myocardium / Q000166:cytology; D020011:Protective Agents / Q000737:chemistry / Q000494:pharmacology*; D011807:Quinolizines / Q000737:chemistry / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D051785:Smad Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D000093842:Matrines
chemical_list: D000450:Aldosterone; D000470:Alkaloids; D020011:Protective Agents; D011807:Quinolizines; D051785:Smad Proteins; C037573:oxymatrine; D000093842:Matrines
keywords: Aldosterone; Cardiac fibroblasts; Oxymatrine; Smad-2; Smad-3; Smad-4