title: Low molecular weight heparin inhibits circulating fibrocytes differentiation by modulating neuronal nitric oxide synthase and TGF-β1/Smad pathway.
abstract: OBJECTIVE
  Circulating fibrocytes (CFs) have been placed at the center of a number of fibrosing conditions. Recently, attention has been drawn to the non-anticoagulant activities of low molecular weight heparin (LH), especially its anti-fibrotic effects. The purpose of this study was to investigate the effects of LH on CFs differentiation and possible underlying mechanisms.
  
  
  RESULTS
  CFs were cultured from human peripheral blood mononuclear cells and identified by dual-immunofluorescence staining. Incubation with LH inhibited CFs trans-differentiation by upregulating CD34 and downregulating pro-Collagen I and a-SMA in a concentration- and time-dependent manner, all of which were detected by flow cytometry. Similar effects were observed after incubation with L-NAME, an inhibitor of NOS. NO production was measured by Griess methods and markedly decreased in CFs treated with LH. Three NOS isoforms were assessed by western blot and nNOS was the predominant isoform involved in this process. Additionally, LH and L-NAME had similar down-regulating effects on the expression of TGF-β1 and pSmad2/3, which indicated that TGF-β/Smad pathway might be a downstream signaling of nNOS/NO during LH treatment.
  
  
  CONCLUSIONS
  These results suggested that LH could exhibit anti-fibrotic effects by inhibiting CFs transdifferentiation, in which the involvement of nNOS/NO and TGF-β/Smad pathway were identified.
authors.affiliation: Department of Cardiovascular Diseases, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Xudong
authors.initials: X
authors.lastname: Xie
authors.forename: Liangfeng
authors.initials: L
authors.lastname: Zhu
authors.forename: Zhongying
authors.initials: Z
authors.lastname: Fu
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Guo
authors.forename: Kan
authors.initials: K
authors.lastname: Wang
authors.forename: Xiaosheng
authors.initials: X
authors.lastname: Hu
authors.forename: Junzhu
authors.initials: J
authors.lastname: Chen
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects*; D048430:Cell Shape; D002478:Cells, Cultured; D005347:Fibroblasts / Q000201:enzymology / Q000502:physiology*; D006495:Heparin, Low-Molecular-Weight / Q000494:pharmacology*; D006801:Humans; D009569:Nitric Oxide / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D015398:Signal Transduction; D051785:Smad Proteins / Q000378:metabolism; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D006495:Heparin, Low-Molecular-Weight; D051785:Smad Proteins; D053773:Transforming Growth Factor beta1; D009569:Nitric Oxide; C496319:NOS2 protein, human; C496314:NOS3 protein, human; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D052250:Nitric Oxide Synthase Type III