PMID- 9974405
OWN - NLM
STAT- MEDLINE
DCOM- 19990316
LR  - 20190831
IS  - 1079-5642 (Print)
IS  - 1079-5642 (Linking)
VI  - 19
IP  - 2
DP  - 1999 Feb
TI  - Expression of tissue inhibitor of metalloproteinase-1, -2, and -3 during 
      neointima formation in organ cultures of human saphenous vein.
PG  - 255-65
AB  - Degradation of the extracellular basement membrane is implicated in 
      atherosclerosis, restenosis after angioplasty, and intimal thickening of vein 
      grafts. Upregulation of metalloproteinase (MMP)-2 and MMP-9 accompanies neointima 
      formation in cholesterol-fed rabbits, in rat and pig models of angioplasty, and 
      in organ cultures of human saphenous veins. MMPs are inhibited by binding to 
      tissue inhibitors of MMPs (TIMPs). Relatively little is known about their 
      regulation in relationship to neointima formation; thus, we investigated TIMP 
      expression in the organ culture model. Qualitative reverse 
      transcriptase-polymerase chain reaction of mRNA extracted from veins showed that 
      TIMP-1, TIMP-2, and TIMP-3 are each expressed before and after culture. 
      Zymography revealed that TIMP-1 was the most abundant TIMP secreted and that its 
      secretion increased dramatically between 0 to 2 and 12 to 14 days of culture. An 
      enzyme-linked immunosorbent assay showed that TIMP-1 secretion increased from 
      3.2+/-1.5 (mean+/-SE) to 32+/-6 ng/mg wet weight per day (n=5, P<0.01). 
      Immunocytochemical testing localized the increased expression of TIMP-1 to 
      neointimal smooth muscle cells. Although less abundant, TIMP-2 secretion also 
      increased from 0.8+/-0. 3 to 4.7+/-0.2 ng/mg wet weight per day (n=5, P<0.001), 
      and tissue levels increased from 33+/-7 to 150+/-70 ng/mg wet weight (P<0.05). 
      TIMP-2 was also immunolocalized to neointimal smooth muscle cells and their 
      surrounding matrix. TIMP-3 was not secreted but was detected variably and 
      constitutively in tissue extracts (160+/-120 and 170+/-100 ng/mg wet weight [n=9] 
      on days 2 and 14, respectively). TIMP-3 was found in the cells and extracellular 
      matrix of the media and adventitia before culture and to a lesser extent in the 
      neointima after 14 days of culture. Rates of total TIMP secretion on day 14 
      exceeded those of MMP-2 and MMP-9 (10.6+/-1.9 and 15.6+/-2.3 ng/mg wet weight per 
      day, respectively). Consistent with this, in situ zymography showed that MMP 
      gelatinase activity was highly localized to cell bodies in the media and 
      neointima. Secretion of TIMP-1 and TIMP-2 is greatly increased during neointima 
      formation in human saphenous veins. TIMP-1 is readily released, whereas TIMP-2 
      remains partially attached and TIMP-3 exclusively attached to the extracellular 
      matrix. Regulation of TIMP expression is therefore an important determinant of 
      net MMP activity during neointima formation, restricting it to the pericellular 
      environment.
FAU - Kranzhofer, A
AU  - Kranzhofer A
AD  - Bristol Heart Institute, University of Bristol, Bristol Royal Infirmary, Bristol, 
      United Kingdom.
FAU - Baker, A H
AU  - Baker AH
FAU - George, S J
AU  - George SJ
FAU - Newby, A C
AU  - Newby AC
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Arterioscler Thromb Vasc Biol
JT  - Arteriosclerosis, thrombosis, and vascular biology
JID - 9505803
RN  - 0 (Culture Media, Conditioned)
RN  - 0 (RNA, Messenger)
RN  - 0 (Tissue Extracts)
RN  - 0 (Tissue Inhibitor of Metalloproteinase-1)
RN  - 0 (Tissue Inhibitor of Metalloproteinase-3)
RN  - 127497-59-0 (Tissue Inhibitor of Metalloproteinase-2)
RN  - EC 3.4.24.- (Collagenases)
RN  - EC 3.4.24.- (Gelatinases)
RN  - EC 3.4.24.- (Metalloendopeptidases)
RN  - EC 3.4.24.24 (Matrix Metalloproteinase 2)
RN  - EC 3.4.24.35 (Matrix Metalloproteinase 9)
SB  - IM
MH  - Collagenases/metabolism
MH  - Culture Media, Conditioned
MH  - Gelatinases/metabolism
MH  - Humans
MH  - Matrix Metalloproteinase 2
MH  - Matrix Metalloproteinase 9
MH  - Metalloendopeptidases/metabolism
MH  - Middle Aged
MH  - Organ Culture Techniques
MH  - RNA, Messenger/metabolism
MH  - Saphenous Vein/metabolism/*physiology
MH  - Tissue Distribution
MH  - Tissue Extracts/metabolism
MH  - Tissue Inhibitor of Metalloproteinase-1/genetics/*metabolism
MH  - Tissue Inhibitor of Metalloproteinase-2/genetics/*metabolism
MH  - Tissue Inhibitor of Metalloproteinase-3/genetics/*metabolism
MH  - Tunica Intima/*physiology
EDAT- 1999/02/12 00:00
MHDA- 1999/02/12 00:01
CRDT- 1999/02/12 00:00
PHST- 1999/02/12 00:00 [pubmed]
PHST- 1999/02/12 00:01 [medline]
PHST- 1999/02/12 00:00 [entrez]
AID - 10.1161/01.atv.19.2.255 [doi]
PST - ppublish
SO  - Arterioscler Thromb Vasc Biol. 1999 Feb;19(2):255-65. doi: 
      10.1161/01.atv.19.2.255.