Tissue inhibitor of metalloproteinase-3 regulates inflammation in human and mouse intestine

Gastroenterology. 2012 Nov;143(5):1277-1287.e4. doi: 10.1053/j.gastro.2012.07.016. Epub 2012 Jul 20.


Background & aims: Tissue inhibitor of metalloproteinases (TIMP)-3 is an inhibitor of matrix metalloproteinases, which regulates tissue inflammation, damage, and repair. We investigated the role of TIMP-3 in intestinal inflammation in human beings and mice.

Methods: We used real-time polymerase chain reaction and flow cytometry to measure levels of TIMP-3 in intestine samples from patients with Crohn's disease (CD) and those without (controls). We also analyzed TIMP-3 levels in lamina propria mononuclear cells (LPMCs) collected from biopsy samples of individuals with or without CD (controls) and then stimulated with transforming growth factor (TGF)-β1, as well as in biopsy samples collected from patients with CD and then incubated with a Smad7 anti-sense oligonucleotide (knock down). LPMCs and biopsy samples from patients with CD were cultured with exogenous TIMP-3 and levels of inflammatory cytokines were measured. We evaluated the susceptibility of wild-type, TIMP-3-knockout (TIMP-3-KO), and transgenic (TIMP-3-Tg) mice to induction of colitis with 2, 4, 6-trinitrobenzene-sulfonic-acid (TNBS), and the course of colitis in recombinase-activating gene-1-null mice after transfer of wild-type or TIMP-3-KO T cells.

Results: Levels of TIMP-3 were reduced in intestine samples from patients with CD compared with controls. Incubation of control LPMCs with TGF-β1 up-regulated TIMP-3; knockdown of Smad7, an inhibitor of TGF-β1, in biopsy samples from patients with CD increased levels of TIMP-3. Exogenous TIMP-3 reduced levels of inflammatory cytokines in CD LPMCs and biopsy samples. TIMP-3-KO mice developed severe colitis after administration of TNBS, whereas TIMP-3-Tg mice were resistant to TNBS-induced colitis. Reconstitution of recombinase-activating gene-1-null mice with T cells from TIMP-3-KO mice increased the severity of colitis, compared with reconstitution with wild-type T cells.

Conclusions: TIMP-3 is down-regulated in inflamed intestine of patients with CD. Its expression is regulated by TGF-β1, and knock-down of Smad7 in intestinal tissues from patient with CD up-regulates TIMP-3. Loss or reduction of TIMP-3 in mice promotes development of colitis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Amyloid Precursor Protein Secretases / metabolism
  • Animals
  • Cells, Cultured
  • Colitis / chemically induced
  • Colitis / genetics
  • Colitis / metabolism
  • Colitis, Ulcerative / metabolism*
  • Crohn Disease / metabolism*
  • Cytokines / metabolism
  • Down-Regulation
  • Gene Knockdown Techniques
  • Gene Knockout Techniques
  • Humans
  • Intestinal Mucosa / drug effects
  • Intestinal Mucosa / metabolism*
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • Mice
  • Mice, Knockout
  • Mice, Transgenic
  • Middle Aged
  • Oligonucleotides, Antisense / pharmacology
  • RNA, Messenger / metabolism*
  • Smad7 Protein / genetics
  • Tissue Inhibitor of Metalloproteinase-3 / genetics
  • Tissue Inhibitor of Metalloproteinase-3 / metabolism*
  • Tissue Inhibitor of Metalloproteinase-3 / pharmacology
  • Transforming Growth Factor beta / pharmacology
  • Trinitrobenzenesulfonic Acid


  • Cytokines
  • Oligonucleotides, Antisense
  • RNA, Messenger
  • Smad7 Protein
  • Tissue Inhibitor of Metalloproteinase-3
  • Transforming Growth Factor beta
  • Trinitrobenzenesulfonic Acid
  • Amyloid Precursor Protein Secretases