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Nitric oxide inhibition increases aortic wall matrix metalloproteinase-9 expression.
J Surg Res. 2002 May 01; 104(1):15-21.JS

Abstract

OBJECTIVE

Nitric oxide (NO) may mediate vessel wall remodeling by regulating expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). This study tested the hypothesis that nitric oxide synthase (NOS) inhibition in whole aortic wall causes increases in cytokine-stimulated MMP and TIMP expression.

METHODS

Cultured infrarenal aortic segments from Sprague-Dawley rats were exposed to increasing concentrations (0, 0.1, 0.5, 1, and 5 mM; n = 6 per concentration) of N(G)-monomethyl-l-arginine (L-NMMA), a known inhibitor of NOS. This was in the presence of 2 ng/ml of interleukin-1beta, a known inducer of NOS, MMP, and TIMP expression. Media nitrate and nitrite (NO(x)) were measured at 72 h using the Saville method. Media MMP activity was measured using gelatin zymography. MMP-2 and -9 protein and mRNA levels were determined by Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). TIMP activity and mRNA levels were evaluated by reverse zymography and RT-PCR. Data were analyzed using ANOVA.

RESULTS

Increasing concentrations of L-NMMA produced a dose-dependent decrease in NO(x) (2214 +/- 405 to 347 +/- 37 ng/mg, P < 0.001). Zymography demonstrated a dose-dependent increase in 92-kDa MMP (pro-MMP-9) activity (P < 0.001) with corresponding increases in pro-MMP-9 protein (P = 0.03) and mRNA levels (P = 0.004). While there was a dose-dependent increase in 72-kDa MMP (pro-MMP-2) activity (P = 0.001), pro-MMP-2 protein and mRNA levels were unchanged. Reverse zymography demonstrated a dose-dependent increase in 29-kDa TIMP-1 activity (P = 0.01), but there was no change in TIMP-1 mRNA levels.

CONCLUSIONS

NOS inhibition in ex vivo aortic tissue causes a dose-dependent increase in MMP-9 expression and activity. It is speculated that deficiencies of NO in vivo alter MMP and TIMP homeostasis, favoring matrix degradation.

Authors+Show Affiliations

Jobst Vascular Research Laboratories, University of Michigan Medical School, Ann Arbor, Michigan, 48109-0329, USA. eagleton@med.umich.eduNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11971672

Citation

Eagleton, Matthew J., et al. "Nitric Oxide Inhibition Increases Aortic Wall Matrix Metalloproteinase-9 Expression." The Journal of Surgical Research, vol. 104, no. 1, 2002, pp. 15-21.
Eagleton MJ, Peterson DA, Sullivan VV, et al. Nitric oxide inhibition increases aortic wall matrix metalloproteinase-9 expression. J Surg Res. 2002;104(1):15-21.
Eagleton, M. J., Peterson, D. A., Sullivan, V. V., Roelofs, K. J., Ford, J. A., Stanley, J. C., & Upchurch, G. R. (2002). Nitric oxide inhibition increases aortic wall matrix metalloproteinase-9 expression. The Journal of Surgical Research, 104(1), 15-21.
Eagleton MJ, et al. Nitric Oxide Inhibition Increases Aortic Wall Matrix Metalloproteinase-9 Expression. J Surg Res. 2002 May 1;104(1):15-21. PubMed PMID: 11971672.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Nitric oxide inhibition increases aortic wall matrix metalloproteinase-9 expression. AU - Eagleton,Matthew J, AU - Peterson,David A, AU - Sullivan,Vita V, AU - Roelofs,Karen J, AU - Ford,John A, AU - Stanley,James C, AU - Upchurch,Gilbert R,Jr PY - 2002/4/25/pubmed PY - 2002/6/12/medline PY - 2002/4/25/entrez SP - 15 EP - 21 JF - The Journal of surgical research JO - J Surg Res VL - 104 IS - 1 N2 - OBJECTIVE: Nitric oxide (NO) may mediate vessel wall remodeling by regulating expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). This study tested the hypothesis that nitric oxide synthase (NOS) inhibition in whole aortic wall causes increases in cytokine-stimulated MMP and TIMP expression. METHODS: Cultured infrarenal aortic segments from Sprague-Dawley rats were exposed to increasing concentrations (0, 0.1, 0.5, 1, and 5 mM; n = 6 per concentration) of N(G)-monomethyl-l-arginine (L-NMMA), a known inhibitor of NOS. This was in the presence of 2 ng/ml of interleukin-1beta, a known inducer of NOS, MMP, and TIMP expression. Media nitrate and nitrite (NO(x)) were measured at 72 h using the Saville method. Media MMP activity was measured using gelatin zymography. MMP-2 and -9 protein and mRNA levels were determined by Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR). TIMP activity and mRNA levels were evaluated by reverse zymography and RT-PCR. Data were analyzed using ANOVA. RESULTS: Increasing concentrations of L-NMMA produced a dose-dependent decrease in NO(x) (2214 +/- 405 to 347 +/- 37 ng/mg, P < 0.001). Zymography demonstrated a dose-dependent increase in 92-kDa MMP (pro-MMP-9) activity (P < 0.001) with corresponding increases in pro-MMP-9 protein (P = 0.03) and mRNA levels (P = 0.004). While there was a dose-dependent increase in 72-kDa MMP (pro-MMP-2) activity (P = 0.001), pro-MMP-2 protein and mRNA levels were unchanged. Reverse zymography demonstrated a dose-dependent increase in 29-kDa TIMP-1 activity (P = 0.01), but there was no change in TIMP-1 mRNA levels. CONCLUSIONS: NOS inhibition in ex vivo aortic tissue causes a dose-dependent increase in MMP-9 expression and activity. It is speculated that deficiencies of NO in vivo alter MMP and TIMP homeostasis, favoring matrix degradation. SN - 0022-4804 UR - https://www.unboundmedicine.com/medline/citation/11971672/Nitric_oxide_inhibition_increases_aortic_wall_matrix_metalloproteinase_9_expression_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022480402963961 DB - PRIME DP - Unbound Medicine ER -