title: Total saponins of panaxnotoginseng promotes lymphangiogenesis by activation VEGF-C expression of lymphatic endothelial cells.
abstract: BACKGROUND
  Lymphatic system plays an important role in maintaining the fluid homeostasis and normal immune responses, anatomic or functional obstruction of which leads to lymphedema, and treatments for therapeutic lymphangiogenesis are efficiency for secondary lymphedema. Total saponins of panaxnotoginseng (PNS) are a mixture isolated from Panaxnotoginseng (Burkill) F.H.Chen, which has been used as traditional Chinese medicine in China for treatment of cardio- and cerebro-vascular diseases. The aim of this study was to determine the effect and mechanism of PNS on lymphangiogenesis.
  
  
  METHODS
  The Tg (fli1: egfp; gata1: dsred) transgenic zebrafish embryos were treated with different concentrations of PNS (10, 50, 100μM) for 48h with or without the 6h pretreatment of the 30μM Vascular endothelial growth factors receptor (VEGFR)-3 kinase inhibitor, followed with morphological observation and lympangiogenesis of thoracic duct assessment. The effect of PNS on cell viability, migration, tube formation and Vascular endothelial growth factors (VEGF)-C mRNA and protein expression of lymphatic endothelial cells (LECs) were determined. The role of phosphatidylinositol-3 (PI-3)-kinase (PI3K), extracellular signal-regulated kinase (ERK)1/2 pathways, c-Jun N-terminal kinase (JNK) and P38 mitogen activated protein kinases (MAPK) signaling in PNS-induced VEGF-C expression of LECs by using pharmacological agents to block each signal.
  
  
  RESULTS
  PNS promotes lymphangiogenesis of thoracic duct in zebrafish with or without VEGFR3 Kinase inhibitor pre-impairment. PNS promotes proliferation, migration and tube formation of LECs. The tube formation induced by PNS could be blocked by VEGFR3 Kinase inhibitor. PNS induce VEGF-C expression of LEC, which could be blocked by ERK1/2, PI3K and P38MAPK signaling inhibitors.
  
  
  CONCLUSIONS
  PNS activates lymphangiogenesis both in vivo and in vitro by up-regulating VEGF-C expression and activation of ERK1/2, PI3K and P38MAPK signaling. These findings provide a novel insight into the role of PNS in lymphangiogenesis and suggest that it might be an attractive and suitable therapeutic agent for treating secondary lymphedema or other lymphatic system impairment related disease.
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: phjod@163.com.
authors.forename: Jinlong
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: 952026015@qq.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: 15221152917@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, NY 14642, USA; Center for Musculoskeletal Research, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, NY 14642, USA. Electronic address: Lianping_xing@urmc.rochester.edu.
authors.forename: Lianping
authors.initials: L
authors.lastname: Xing
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: hoxu@163.com.
authors.forename: Hao
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: yjwang8888@126.com.
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: jzyjs200032@126.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Shi
authors.affiliation: Department of Orthopaedics, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China. Electronic address: liangqianqiantcm@126.com.
authors.forename: Qianqian
authors.initials: Q
authors.lastname: Liang
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002460:Cell Line; D042783:Endothelial Cells / Q000201:enzymology / Q000378:metabolism*; D042583:Lymphangiogenesis / Q000187:drug effects*; D020935:MAP Kinase Signaling System; D051379:Mice; D053723:Panax notoginseng / Q000737:chemistry*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D012503:Saponins / Q000494:pharmacology*; D015398:Signal Transduction; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D012503:Saponins; D042582:Vascular Endothelial Growth Factor C; D019869:Phosphatidylinositol 3-Kinases; D048051:p38 Mitogen-Activated Protein Kinases
keywords: Acrylamide (PubChemCID: 6579); Ammonium Persulfate (PubChemCID:62648); Chloroform (PubChemCID:6212); Dimethyl sulfoxide (DMSO) (PubChem CID: 679); Ethanol (PubChem CID: 702); Glycine (PubChemCID:750); Lymphangiogenesis; Lymphedema; MAZ51, VEGF Receptor 3 Kinase inhibitor (PubChemCID:9839842); Methanol (PubChemCID:887); PD98059 (PubChemCID: 4713); SB203580 (PubChemCID: 176155); SP600125 (PubChemCID: 8515); Saponins of panaxnotoginseng; Sodium chloride (PubChemCID:5234); Vascular endothelial growth factors C; Wortmannin (PubChemCID: 312145)