Reappraisal of Quantitative Gel Zymography for Matrix Metalloproteinases

Prescimone, Tommaso; Tognotti, Danika; Caselli, Chiara; Cabiati, Manuela; D'Amico, Andrea; Ry, Silvia Del; Giannessi, Daniela

doi:10.1002/jcla.21696

Cited by 8 publications

(5 citation statements)

References 44 publications

(56 reference statements)

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“…As in previous reports, we found a strong correlation ( r = 0.81) between MMP-9 active and total form measured, respectively, by gel zymography and ELISA 38 , 39 . However, the Pearson’s correlation coefficient ( r ) does not assess agreement between the two methods but rather measures the association between them.…”

Section: Discussionsupporting

confidence: 90%

“…Specifically, Prescimone et al, compared both methods in 25 volunteers using correlation coefficient and Bland–Altman difference analysis. Based on the correlation coefficient and regardless of the absolute systematic error (0.006 mg/l) revealed by Bland–Altman analysis, they concluded that both methods offer similar reliability for quantifying MMP-9 38 . In accordance with their results, we obtained a positive bias between gel zymography and ELISA, but on a significant larger scale (0.11 mg/l).…”

Section: Discussionmentioning

confidence: 99%

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Clinical significance of matrix metalloproteinase-9 in Fragile X Syndrome

Laroui

Galarneau

Abolghasemi

et al. 2022

Sci Rep

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High plasma matrix metalloproteases-9 (MMP-9) levels have been reported in Fragile X Syndrome in a limited number of animal and human studies. Since the results obtained are method-dependent and not directly comparable, the clinical utility of MMP-9 measurement in FXS remains unclear. This study aimed to compare quantitative gel zymography and ELISA and to determine which method better discriminates abnormal MMP-9 levels of individuals with FXS from healthy controls and correlates with the clinical profile. The active and total forms of MMP-9 were quantified respectively, by gel zymography and ELISA in a cohort of FXS (n = 23) and healthy controls (n = 20). The clinical profile was assessed for the FXS group using the Aberrant Behavior Checklist FXS adapted version (ABC-CFX), Adaptive Behavior Assessment System (ABAS), Social Communication Questionnaire (SCQ), and Anxiety Depression and Mood Scale questionnaires. Method comparison showed a disagreement between gel zymography and ELISA with a constant error of − 0.18 [95% CI: − 0.35 to − 0.02] and a proportional error of 2.31 [95% CI: 1.53 to 3.24]. Plasma level of MMP-9 active form was significantly higher in FXS (n = 12) as compared to their age-sex and BMI matched controls (n = 12) (p = 0.039) and correlated with ABC-CFX (rs = 0.60; p = 0.039) and ADAMS (rs = 0.57; p = 0.043) scores. As compared to the plasma total form, the plasma MMP-9 active form better enables the discrimination of individuals with FXS from controls and correlates with the clinical profile. Our results highlight the importance of choosing the appropriate method to quantify plasma MMP-9 in future FXS clinical studies.

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Section: Discussionsupporting

confidence: 90%

Section: Discussionmentioning

confidence: 99%

Clinical significance of matrix metalloproteinase-9 in Fragile X Syndrome

Laroui

Galarneau

Abolghasemi

et al. 2022

Sci Rep

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“…The zymography is a reproducible and reliable method to detect gelatinase activity (Prescimone et al, ). It is based on sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) with the 0.5% gelatin (type A from porcine skin, Sigma‐Aldrich, St Louis, MO) as substrate for detection of MPs enzymatic activity.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of Metalloproteinase Activity in FANCA Is Linked to Altered Oxygen Metabolism

Ravera

Capanni

Tognotti

et al. 2014

Journal Cellular Physiology

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Bone marrow (BM) failure, increased risk of myelodysplastic syndrome, acute leukaemia and solid tumors, endocrinopathies and congenital abnormalities are the major clinical problems in Fanconi anemia patients (FA). Chromosome instability and DNA repair defects are the cellular characteristics used for the clinical diagnosis. However, these biological defects are not sufficient to explain all the clinical phenotype of FA patients. The known defects are structural alteration in cell cytoskeleton, altered structural organization for intermediate filaments, nuclear lamina, and mitochondria. These are associated with different expression and/or maturation of the structural proteins vimentin, mitofilin, and lamin A/C suggesting the involvement of metalloproteinases (MPs). Matrix metalloproteinases (MMP) are involved in normal physiological processes such as human skeletal tissue development, maturation, and hematopoietic reconstitution after bone marrow suppression. Current observations upon the eventual role of MPs in FA cells are largely inconclusive. We evaluated the overall MPs activity in FA complementation group A (FANCA) cells by exposing them to the antioxidants N-acetyl cysteine (NAC) and resveratrol (RV). This work supports the hypothesis that treatment of Fanconi patients with antioxidants may be important in FA therapy.

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“…The use of MMP and TIMP plasma levels as biomarkers in different clinical settings is increasing due to their diagnostic and prognostic value in human medicine (23,27). Currently, only a few studies have been focused on plasma or milk levels of MMP (21,28), while the plasma level of TIMPs in cattle is lack of relevant research.…”

Section: Discussionmentioning

confidence: 99%

Change in Proteolytic Profile in Heifers After Oligofructose Overload

Zheng

Ding

et al. 2020

Front. Vet. Sci.

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Laminitis in cattle is an important underlying cause of lameness, which leads to a significant reduction in economic and animal welfare. Nevertheless, the disordered pathological processes of laminitis remain unclear. Several proteinases are probably involved in the disorder of basement membrane (BM) metabolism in laminitis, for instance, matrix metalloproteinases (MMPs), neutrophil elastase (NE), and myeloperoxidase (MPO). This study aimed to investigate the change in proteolytic profile in circulating and lamellar tissues using an oligofructose (OF) overload-induced laminitis model in heifers. Twelve clinically healthy and nonlame Chinese Holstein heifers were recruited and randomly divided into two groups: OF-induced and control (CON). The OF-induced heifers group (n = 6) was administered 17 g/kg of body weight (BW) of OF dissolved in 2 L/100 kg of BW of tap water via the oral-rumen tube. The CON group (n = 6) was given an equal volume of tap water. The plasma samples were collected 0, 6, 12, 18, 24, 36, 48, 60, and 72 h after administration, and the lamellar samples were collected 72 h after euthanasia. The plasma samples were analyzed by zymography and reverse zymography. Histological examination, zymography, reverse zymography, and Western blot of lamellar samples were conducted. In the plasma of the OF-induced group, the pro-MMP9 activity increased from 36 h (P < 0.001) to 60 h (P < 0.05). Moreover, the plasma tissue inhibitors of metalloproteinase 1 (TIMP1) activity decreased after 18 h (P < 0.05), while the ratio of pro-MMP9 to TIMP1 and TIMP2 increased after 18 h (P < 0.001) and 48 h (P < 0.05), respectively. The act-MMP2, pro-MMP9, and act-MMP9 activities increased in the lamellar tissue of the OF-induced group compared with the CON group (P < 0.05). In addition, the expression of lamellar NE protein was higher in the OF-induced group (P < 0.01), while no change was found in lamellar MPO protein compared with the CON group. In conclusion, increased pro-MMP9 combined with decreased TIMP1 activity in the circulation might have caused the activation of blood neutrophils, while the activation of proteolytic enzymes in lamellar tissue probably led to the dysfunction of BM in the OF-induced group.

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Reappraisal of Quantitative Gel Zymography for Matrix Metalloproteinases

Cited by 8 publications

References 44 publications

Clinical significance of matrix metalloproteinase-9 in Fragile X Syndrome

Clinical significance of matrix metalloproteinase-9 in Fragile X Syndrome

Inhibition of Metalloproteinase Activity in FANCA Is Linked to Altered Oxygen Metabolism

Change in Proteolytic Profile in Heifers After Oligofructose Overload

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