title: [Co-overexpression of human tissue kallikrein 1 and human metalloproteinase 1 tissue inhibitor inhibits neointima formation in the rat artery after balloon angioplasty].
abstract: OBJECTIVE
  To investigate the effects of adenovirus-mediated human tissue kallikrein 1(hTK-1) and/or human tissue metalloproteinase inhibitor 1 (hTIMP-1) gene delivery on the neointima formation in balloon-injured rat carotids and related mechanism.
  
  
  METHODS
  Forty-six male Sprague-Dawley rats were randomly assigned into 6 groups with the random number table: (1) sham-operated group(n=6), (2) angioplasty group (n=8), (3) vector virus group (n=8), (4) hTK-1 group (n=8), (5) hTIMP-1 group (n=8), (6) hTK-1-hTIMP-1 group (n=8). Except sham rats, all rats underwent carotid artery balloon injury and local delivery of saline or different recombined adenoviruses respectively. Rats were sacrificed 14 days later. Intima/media area ratio was assessed on hematoxylin-eosin stained tissue section. Immunofluorescence images stained for hTK-1, hTIMP-1 were obtained and analyzed by the confocal microscope for co-localization examination of hTK-1 and hTIMP-1. The protein expression levels of hTK-1, hTIMP-1, matrix metalloproteinases(MMP)-2 and MMP-9 were determined by Western blot. Immune histochemical staining for PCNA was also performed.
  
  
  RESULTS
  (1)Intima area, intima/media area ratio, PCNA, MMP-2 and MMP-9 levels were all significantly increased in rats underwent angioplasty (did or did not receive vector virus) compared with sham-operated rats (all P<0.01) while above parameters were similar between rats underwent angioplasty or vector virus delivery (all P>0.05). (2) The intima area of rats received vector virus, hTK-1, hTIMP-1 or dual gene transfer were (0.160±0.010), (0.110±0.015), (0.121±0.016) or (0.081±0.008) mm(2) respectively, intima area was similar between rats received hTK-1 or hTIMP-1 (P>0.05), differences were found between other groups (all P<0.01). The intima/media area ratio of rats received vector virus, hTK-1, hTIMP-1 or dual gene transfer were 2.035±0.117, 1.443±0.097, 1.522±0.078 or 0.972±0.072 respectively, no difference was found between rats received hTK-1 or hTIMP-1 in intima/media area ratio (all P>0.05), differences were found between other groups (all P<0.01). The MMP-2 and MMP-9 expression of rats received vector virus, hTK-1, hTIMP-1 or dual gene transfer were 0.817±0.036, 0.606±0.044, 0.571±0.061 or 0.455±0.030 and 0.745±0.057, 0.613±0.038, 0.582±0.050 or 0.473±0.038 respectively, no difference was found between rats received hTK-1 or hTIMP-1 in MMP-2 or MMP-9 expression (all P>0.05), differences were found between other groups (all P<0.01). The PCNA expression of rats received vector virus, hTK-1, hTIMP-1 or dual gene transfer were 0.065±0.007, 0.052±0.004, 0.055±0.007 or 0.031±0.004 respectively, no difference was found between rats received hTK-1or hTIMP-1 in PCNA expression (all P>0.05), differences were found between other groups (all P<0.01).
  
  
  CONCLUSIONS
  hTK-1 and hTIMP-1 co-overexpression may synergistically inhibit neointimal hyperplasia, attenuate vascular remodeling and reduce restenosis possibly via down regulating the expressions of PCNA, MMP-2 and MMP-9 in balloon-injured rat carotids.
authors.affiliation: Department of Geriatrics, Fujian Provincial Hospital, Fujian Provincial Institute of Clinical Geriatrics, Provincial Clinical Medical College of Fujian Medical University, Fuzhou 350001, China.
authors.forename: H Z
authors.initials: HZ
authors.lastname: Yu
authors.forename: F
authors.initials: F
authors.lastname: Zhang
authors.forename: P L
authors.initials: PL
authors.lastname: Zhu
authors.forename: W
authors.initials: W
authors.lastname: Pan
authors.forename: S J
authors.initials: SJ
authors.lastname: Huang
authors.forename: H
authors.initials: H
authors.lastname: Xiang
mesh_terms: D000256:Adenoviridae; D000800:Angioplasty, Balloon; D000818:Animals; D020212:Carotid Artery Injuries / Q000628:therapy*; D017536:Carotid Artery, Common / Q000473:pathology*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D006965:Hyperplasia / Q000473:pathology; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D058426:Neointima / Q000517:prevention & control*; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000235:genetics / Q000378:metabolism*; D020840:Tissue Kallikreins / Q000235:genetics / Q000378:metabolism*
chemical_list: C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; D020840:Tissue Kallikreins; D020778:Matrix Metalloproteinase 2; C522363:Mmp2 protein, rat; D020780:Matrix Metalloproteinase 9; C572337:Mmp9 protein, rat