title: Tongxinluo promotes mesenchymal stem cell tube formation in vitro.
abstract: OBJECTIVE
  To study whether Tongxinluo (TXL) can induce angiogenesis in bone marrow mesenchymal stem cells (MSCs), and to investigate the underlying mechanism.
  
  
  METHODS
  Bone marrow MSCs were obtained from male Sprague-Dawley rats. We established an angiogenesis model in vitro via matrigel experiment. MSCs were seeded on matrigel coated 24-well plates, and treated by TXL 50 and 100 mg/L. After 24 h, we observed the tube formations of MSCs in the matrigel. Cell migration ability was examined by wound scratch test and transwell assay. Expressions of vascular endothelial growth factor (VEGF), fetal liver kinase-1 (Flk-1), matrix metalloproteinase-2 (MMP-2), MMP-9, and tissue inhibitor of metalloproteinase-2 (TIMP-2) were analyzed at the protein level by Western blot. Gelatin zymography assay was applied to investigating the MSC paracrine abilities of pro-MMP-2 and activated MMP-2.
  
  
  RESULTS
  TXL promoted MSC tube formation in matrigel. The ratio of TXL 100 mg/L treated-MSC tubular length was increased 3.04-fold compared to the control group (P<0.05). Scratch test and transwell assay showed that TXL could improve the cell migration ability of MSCs. Western blot experiments showed that TXL promoted MSC synthesis of MMP-2, but it had no influence on the expressions of MMP-9 and TIMP-2. This effect was confirmed by gelatin zymography assay, which showed that TXL increased MSC secretion of pro-MMP-2 and activated MMP-2. VEGF expression of TXL treated-MSCs was increased compared to the control group. The expression of Flk-1 was not different among the groups.
  
  
  CONCLUSIONS
  This study demonstrates that TXL can promote the tube formation of MSCs, and the underlying mechanisms are associated with increased migration ability of MSCs and the up-regulation of MMP-2 and VEGF expressions.
authors.affiliation: Cardiovascular Key Lab of Zhejiang Province, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
authors.forename: Xin-Yang
authors.initials: XY
authors.lastname: Hu
authors.forename: Wen-Xia
authors.initials: WX
authors.lastname: Wang
authors.forename: Min-Jia
authors.initials: MJ
authors.lastname: Yu
authors.forename: Xian-Bao
authors.initials: XB
authors.lastname: Liu
authors.forename: Rong-Rong
authors.initials: RR
authors.lastname: Wu
authors.forename: Feng
authors.initials: F
authors.lastname: Gao
authors.forename: Xin
authors.initials: X
authors.lastname: Huang
authors.forename: Jiang
authors.initials: J
authors.lastname: Cao
authors.forename: Xiao-Jie
authors.initials: XJ
authors.lastname: Xie
authors.forename: Jian-An
authors.initials: JA
authors.lastname: Wang
mesh_terms: D000818:Animals; D002465:Cell Movement; D003094:Collagen / Q000737:chemistry; D004338:Drug Combinations; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D005786:Gene Expression Regulation; D066298:In Vitro Techniques; D007797:Laminin / Q000737:chemistry; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D009389:Neovascularization, Pathologic; D018919:Neovascularization, Physiologic; D011509:Proteoglycans / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D014945:Wound Healing
chemical_list: D004338:Drug Combinations; D004365:Drugs, Chinese Herbal; D007797:Laminin; D011509:Proteoglycans; D042461:Vascular Endothelial Growth Factor A; C513941:tongxinluo; C059485:matrigel; D003094:Collagen; D020778:Matrix Metalloproteinase 2