medigraphic.com
ISSN 2448-8747 (Print)
Publicación cuatrimestral editada por la Secretaría de Salud Jalisco

2022, Number Esp

<< Back Next >>

Sal Jal 2022; 9 (Esp)

Actividad de las enzimas de la matriz extracelular 2 y 9 en leche humana, un estudio exploratorio

Villa-Soto A, Sampieri-Ramírez CL, Arrazate-García MC, Sandoval-Lozano VH, Cocotle-Ronzón Y, Soto-Ojeda GA, Zamora-Bello I

Language: Spanish
References: 19
Page: 45-51
PDF size: 226.61 Kb.


ABSTRACT

Objective: to compare by gelatin zymography the enzymatic activity of MMP-2 and MMP-9 in human milk samples from mothers with infants less than six months of age and infants equal to or greater than six months of age. Material and methods: non-randomized exploratory observational study that included the signing of an informed consent. Acrylamide gels copolymerized with 1 mg/mL gelatin incubated with CaCl2 were used to determine enzymatic activity and 8.5% acrylamide gels to verify the integrity of the samples. Copolymerized gels with 1 mg/mL gelatin incubated with EDTA were used as negative control for enzymatic activity. Images were taken using a photodocumentator. Visual analysis of enzyme activity was performed, and the extent of enzyme activity was quantifi ed using Image J v1.4r soft ware. Results: enzymatic activity of MMP-2 (72 KDa) and MMP-9 (225 KDa, 130 KDa and 92 KDa) was detected in human milk samples. There were no diff erences between the enzymatic activity of MMP-2 and MMP-9 between the groups of mothers with babies less than six months of age compared to the group of mothers with babies equal to or greater than six months of age. Conclusions: the gelatin zymography technique is useful for detecting gelatinase activity of MMP-2 and MMP-9 in human milk samples. This study is a pioneer in demonstrating enzymatic activity of MMP-2 and MMP-9 in samples of human milk from mothers with babies equal to or greater than six months of age.


REFERENCES

  1. Brew K, Dinakarpandian D, Nagase H. Tissue inhibitors ofmetalloproteinases: Evolution, structure and function. BiochimBiophys Acta - Protein Struct Mol Enzymol. 2000;1477(1–2):267–83.

  2. Ganea E, Trifan M, Laslo AC, Putina G, Cristescu C. Matrixmetalloproteinases: Useful and deleterious. Biochem Soc Trans.2007;35(4):689–91.

  3. Woessner JF. Matrix metalloproteinases and their inhibitors inconnective tissue remodeling. FASEB J. 1991;5(8):2145–54.

  4. González-Ávila G, González A, Delgado J, Gutiérrez-GonzálesLH. Participación de las metaloproteasas de matriz en laprogresión del cáncer. Rev del Isntituto Nac EnfermedadesRespir. 2010;22(4):328–36.

  5. Lubetzky R, Mandel D, Mimouni FB, Herman L, Reich R, Reif S.MMP-2 and MMP-9 and their tissue inhibitor in preterm humanmilk. J Pediatr Gastroenterol Nutr. 2010;51(2):210–2.

  6. Coronato S, Laguens G, Di Girolamo V. Rol de lasmetaloproteinasas y sus inhibidores en patología tumoral. Med.2012;72(6):495–502.

  7. Leber TM, Balkwill FR. Zymography: A single-step stainingmethod for quantitation of proteolytic activity on substrate gels.Anal Biochem. 1997;249(1):24–8.

  8. Toth M, Fridman R. Gelatin Zymography. Defi nitions.2020;57:163–74.

  9. Mora J, Manzur AJ, Ramírez T, Silva D. Papel de lasMetaloproteasas de la Matriz en la Degradación del Tejido Pulpar.2005;Vol. 1.

  10. Urbina, Alvaro M. COMPARACIÓN DE LA ACTIVIDADGELATINÁSICA A Y B (MMP-2 Y MMP-9) EN EL LÍQUIDOSINOVIAL PROVENIENTE DE LA ARTICULACIÓNMETACARPOFALÁNGICA EQUINA NORMAL YALTERADA”. Bosque. 2005;

  11. Chirico G, Marzollo R, Cortinovis S, Fonte C, Gasparoni A.Antiinfective properties of human milk. J Nutr. 2008;138(9):1801–6.

  12. Castillo Belén JR, Rams Veranes A, Castillo Belén A, RizoRodríguez R, Cádiz Lahens A. Lactancia materna e inmunidad:impacto social. Medisan. 2009;13(1):0–0.

  13. Cheung PY, Sawicki G, Gross S, Van Aerde J, Radomski M.Diff erential expression of matrix metalloproteinases and thetissue inhibitor in human milk. Proc West Pharmacol Soc.2001;44(May 2014):97–8.

  14. Schanler R. Human milk for preterm infants: nutritional andimmune factors - PubMed [Internet]. 1989 [cited 2021 Feb 20].Available from: https://pubmed.ncbi.nlm.nih.gov/2662415/

  15. Metallinou D, Lykeridou K, Karampas G, Liosis GT, SkevakiC, Rizou M, et al. Postpartum human breast milk levels ofneutrophil gelatinase-associated lipocalin (NGAL) and matrixmetalloproteinase-9 (MMP-9)/NGAL complex in normal andpregnancies complicated with insulin-dependent gestationaldiabetes mellitus. A prospective pilot ca. J Obstet Gynaecol(Lahore) [Internet]. 2020;40(4):461–7. Available from: https://doi.org/10.1080/01443615.2019.1628191

  16. Makowski GS, Ramsby ML. Calibrating gelatin zymograms withhuman gelatinase standards. Anal Biochem. 1996;236(2):353–6.

  17. Sandoval V. Actividad de las enzimas de la matriz extracelular2 y 9 en muestras clínicas de orina determinada por zimografíaen pacientes con diabetes mellitus tipo 2 con y sin enfermedadrenal crónica. 2018;1–60. Available from: https://cdigital.uv.mx/bitstream/handle/123456789/46427/QuirozCortesMCarmen.pdf?sequence=2&isAllowed=y

  18. Schulz CG, Sawicki G, Lemke RP, Roeten BM, Schulz R, CheungPY. MMP-2 and MMP-9 and Th eir Tissue Inhibitors in the Plasmaof Preterm and Term Neonates. Pediatr Res. 2004;55(5):794–801.

  19. Dallas DC, Murray NM, Gan J. Proteolytic Systems in Milk:Perspectives on the Evolutionary Function within the MammaryGland and the Infant. J Mammary Gland Biol Neoplasia. 2015;20(3–4):133–47.




2020     |     www.medigraphic.com