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Balance between matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in the cervical mucus plug estimated by determination of free non-complexed TIMP.
Reprod Biol Endocrinol. 2008 Sep 30; 6:45.RB

Abstract

BACKGROUND

The cervical mucus plug (CMP) is a semi-solid structure with antibacterial properties positioned in the cervical canal during pregnancy. The CMP contains high concentrations of matrix metalloproteinase 8 and 9 (MMP-8, MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1). This indicates a potential to degrade extracellular matrix components depending on the balance between free non-complexed inhibitors and active enzymes.

METHODS

Thirty-two CMPs collected during active labor at term were analyzed. Twelve CMPs were separated into a cellular and an extracellular/fluid phase and analyzed by gelatin and reverse zymography to reveal MMP and TIMP location. Twenty samples were homogenized, extracted and studied by the TIMP activity assay based on gelatin zymography. Enzyme-linked immunosorbent assay (ELISA) was used to determine TIMP-1, MMP-8 and MMP-9 protein concentrations, and gelatin and reverse zymography used to identify gelatinases and TIMPs, respectively. The Western blotting technique was applied for semi-quantification of alpha2-macroglobulin. An ELISA activity assay was used to detect MMP-8 and MMP-9 activity.

RESULTS

ProMMP-2, proMMP-9, TIMP-1 and TIMP-2 were almost exclusively located in the fluid phase compared to the cellular phase of the CMP. All the extracted samples contained MMP-8, MMP-9, TIMP-1, TIMP-2 and alpha2-macroglobulin. Free non-complexed TIMP was detected in all the samples analyzed by the TIMP activity assay and was associated with TIMP-1 protein (R = 0.71, p < 0.001) and with the TIMP/MMP molar ratio (1.7 (1.1-2.5) (mean (95% confidence interval)) (R = 0.65, p = 0.002). The ELISA activity assay showed no activity from MMP-8 or MMP-9.

CONCLUSION

Due to their extracellular location, potential proteolytic activity from neutrophil-derived MMPs in the CMP could exert a biological impact on cervical dilatation and fetal membrane rupture at term. The functional TIMP activity assay, revealing excess non-complexed TIMP, and a molar inhibitor/enzyme ratio above unity, indicate that refined MMP control prevents CMP-originated proteolytic activity in the surrounding tissue.

Authors+Show Affiliations

Department of Obstetrics and Gynecology, Arhus University Hospital, Skejby, Aarhus N, Denmark. nbecher@dadlnet.dkNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

18826601

Citation

Becher, Naja, et al. "Balance Between Matrix Metalloproteinases (MMP) and Tissue Inhibitors of Metalloproteinases (TIMP) in the Cervical Mucus Plug Estimated By Determination of Free Non-complexed TIMP." Reproductive Biology and Endocrinology : RB&E, vol. 6, 2008, p. 45.
Becher N, Hein M, Uldbjerg N, et al. Balance between matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in the cervical mucus plug estimated by determination of free non-complexed TIMP. Reprod Biol Endocrinol. 2008;6:45.
Becher, N., Hein, M., Uldbjerg, N., & Danielsen, C. C. (2008). Balance between matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in the cervical mucus plug estimated by determination of free non-complexed TIMP. Reproductive Biology and Endocrinology : RB&E, 6, 45. https://doi.org/10.1186/1477-7827-6-45
Becher N, et al. Balance Between Matrix Metalloproteinases (MMP) and Tissue Inhibitors of Metalloproteinases (TIMP) in the Cervical Mucus Plug Estimated By Determination of Free Non-complexed TIMP. Reprod Biol Endocrinol. 2008 Sep 30;6:45. PubMed PMID: 18826601.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Balance between matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in the cervical mucus plug estimated by determination of free non-complexed TIMP. AU - Becher,Naja, AU - Hein,Merete, AU - Uldbjerg,Niels, AU - Danielsen,Carl Christian, Y1 - 2008/09/30/ PY - 2008/07/08/received PY - 2008/09/30/accepted PY - 2008/10/2/pubmed PY - 2009/1/8/medline PY - 2008/10/2/entrez SP - 45 EP - 45 JF - Reproductive biology and endocrinology : RB&E JO - Reprod Biol Endocrinol VL - 6 N2 - BACKGROUND: The cervical mucus plug (CMP) is a semi-solid structure with antibacterial properties positioned in the cervical canal during pregnancy. The CMP contains high concentrations of matrix metalloproteinase 8 and 9 (MMP-8, MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1). This indicates a potential to degrade extracellular matrix components depending on the balance between free non-complexed inhibitors and active enzymes. METHODS: Thirty-two CMPs collected during active labor at term were analyzed. Twelve CMPs were separated into a cellular and an extracellular/fluid phase and analyzed by gelatin and reverse zymography to reveal MMP and TIMP location. Twenty samples were homogenized, extracted and studied by the TIMP activity assay based on gelatin zymography. Enzyme-linked immunosorbent assay (ELISA) was used to determine TIMP-1, MMP-8 and MMP-9 protein concentrations, and gelatin and reverse zymography used to identify gelatinases and TIMPs, respectively. The Western blotting technique was applied for semi-quantification of alpha2-macroglobulin. An ELISA activity assay was used to detect MMP-8 and MMP-9 activity. RESULTS: ProMMP-2, proMMP-9, TIMP-1 and TIMP-2 were almost exclusively located in the fluid phase compared to the cellular phase of the CMP. All the extracted samples contained MMP-8, MMP-9, TIMP-1, TIMP-2 and alpha2-macroglobulin. Free non-complexed TIMP was detected in all the samples analyzed by the TIMP activity assay and was associated with TIMP-1 protein (R = 0.71, p < 0.001) and with the TIMP/MMP molar ratio (1.7 (1.1-2.5) (mean (95% confidence interval)) (R = 0.65, p = 0.002). The ELISA activity assay showed no activity from MMP-8 or MMP-9. CONCLUSION: Due to their extracellular location, potential proteolytic activity from neutrophil-derived MMPs in the CMP could exert a biological impact on cervical dilatation and fetal membrane rupture at term. The functional TIMP activity assay, revealing excess non-complexed TIMP, and a molar inhibitor/enzyme ratio above unity, indicate that refined MMP control prevents CMP-originated proteolytic activity in the surrounding tissue. SN - 1477-7827 UR - https://www.unboundmedicine.com/medline/citation/18826601/Balance_between_matrix_metalloproteinases__MMP__and_tissue_inhibitors_of_metalloproteinases__TIMP__in_the_cervical_mucus_plug_estimated_by_determination_of_free_non_complexed_TIMP_ L2 - https://rbej.biomedcentral.com/articles/10.1186/1477-7827-6-45 DB - PRIME DP - Unbound Medicine ER -