title: Alkaloid rich fraction from Nelumbo nucifera targets VSMC proliferation and migration to suppress restenosis in balloon-injured rat carotid artery.
abstract: OBJECTIVE
  Restenosis- an adverse consequence following angioplasty, and atherosclerosis are characterized by abnormal vascular smooth muscle cell (VSMC) proliferation and migration leading to neo-intima formation. In the present study, we investigated the inhibitory effects of alkaloid rich fraction (ARF) from Nelumbo nucifera and isolated compound neferine on platelet-derived growth factor (PDGF-BB) induced VSMC proliferation and migration in vitro and neo-intima formation in a rat carotid artery injury model.
  
  
  METHODS
  PDGF-BB induced VSMC proliferation and migration was assessed using colorimetric assay and modified Boyden chamber method respectively. Gene expression of cell cycle associated molecules was determined by reverse transcription-polymerase chain reaction (RT-PCR). The signaling molecules such as PDGF-Rβ, extracellular regulated kinase (ERK)1/2, c-Jun N-terminal kinase (JNK), P38, metalloproteinase (MMP)-9 and nuclear factor-kappa B (NF-κB) were determined by western blot analysis. Stress fiber formation was evaluated using immunofluorescence microscopy. The rat carotid artery balloon injury model was performed to assess the effect of ARF on neo-intima formation.
  
  
  RESULTS
  ARF possessed the strongest anti-oxidant activities. The anti-proliferative activity of both ARF and neferine was due to suppression of cyclin D1, cyclin E and cyclin-dependent kinase (Cdk) gene expression. Moreover, ARF and neferine inhibited PDGF-Rβ, ERK1/2, JNK and P38 activations and NF-κB translocation. Also, ARF and neferine inhibited VSMC migration by inhibiting MMP-9 activity without affecting cytoskeleton remodeling. In a rat carotid artery injury model, ARF inhibited neo-intima formation.
  
  
  CONCLUSIONS
  Our results indicate that ARF targets VSMC proliferation and migration to attenuate neo-intima formation by inhibition of PDGF-Rβ mediated signaling.
authors.affiliation: Department of Oriental Medicine Resources, Mokpo National University, Muan-gun, Jeonnam 534-729, Republic of Korea.
authors.forename: Moon Young
authors.initials: MY
authors.lastname: Jun
authors.affiliation: Department of Oriental Medicine Resources, Mokpo National University, Muan-gun, Jeonnam 534-729, Republic of Korea.
authors.forename: Rajendra
authors.initials: R
authors.lastname: Karki
authors.affiliation: Department of Oriental Medicine Resources, Mokpo National University, Muan-gun, Jeonnam 534-729, Republic of Korea.
authors.forename: Keshav Raj
authors.initials: KR
authors.lastname: Paudel
authors.affiliation: Department of Oriental Medicine Resources, Mokpo National University, Muan-gun, Jeonnam 534-729, Republic of Korea.
authors.forename: Bhesh Raj
authors.initials: BR
authors.lastname: Sharma
authors.affiliation: East West College of Pharmacy, Bangalore, Karnataka, India.
authors.forename: Deepak
authors.initials: D
authors.lastname: Adhikari
authors.affiliation: Department of Oriental Medicine Resources, Mokpo National University, Muan-gun, Jeonnam 534-729, Republic of Korea. Electronic address: mokpou@yahoo.co.kr.
authors.forename: Dong-Wook
authors.initials: DW
authors.lastname: Kim
mesh_terms: D000470:Alkaloids / Q000737:chemistry*; D000800:Angioplasty, Balloon / Q000009:adverse effects*; D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D000077214:Becaplermin; D001713:Biphenyl Compounds / Q000737:chemistry; D002339:Carotid Arteries / Q000378:metabolism; D020212:Carotid Artery Injuries / Q000378:metabolism; D016893:Carotid Stenosis / Q000517:prevention & control*; D002465:Cell Movement; D049109:Cell Proliferation; D002614:Chelating Agents / Q000737:chemistry; D016166:Free Radical Scavengers / Q000737:chemistry; D015227:Lipid Peroxidation; D020935:MAP Kinase Signaling System; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects*; D016328:NF-kappa B / Q000378:metabolism; D031653:Nelumbo / Q000737:chemistry*; D058426:Neointima / Q000473:pathology; D010851:Picrates / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry; D020574:Proto-Oncogene Proteins c-sis / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000378:metabolism
chemical_list: D000470:Alkaloids; D000975:Antioxidants; D001713:Biphenyl Compounds; D002614:Chelating Agents; D016166:Free Radical Scavengers; D016328:NF-kappa B; D010851:Picrates; D010936:Plant Extracts; D020574:Proto-Oncogene Proteins c-sis; D000077214:Becaplermin; C004931:1,1-diphenyl-2-picrylhydrazyl; D020797:Receptor, Platelet-Derived Growth Factor beta
keywords: Atherosclerosis; Migration; Neferine; Nelumbo nucifera; Proliferation; Restenosis; VSMC