Lupinus albus Protein Components Inhibit MMP-2 and MMP-9 Gelatinolytic Activity In Vitro and In Vivo.

Abstract

Matrix metalloproteinases 2 and 9 (MMP-2 and MMP-9) are regarded as important clinical targets due to their nodal-point role in inflammatory and oncological diseases. Here, we aimed at isolating and characterizing am MMP-2 and-9 inhibitor (MMPI) from Lupinus albus and at assessing its efficacy in vitro and in vivo. The protein was isolated using chromatographic and 2-D electrophoretic procedures and sequenced by using MALDI-TOF TOF and MS/MS analysis. In vitro MMP-2 and 9 inhibitions were determined on colon adenocarcinoma (HT29) cells, as well as by measuring the expression levels of genes related to these enzymes. Inhibitory activities were also confirmed in vivo using a model of experimental TNBS-induced colitis in mice, with oral administrations of 15 mg·kg-1. After chromatographic and electrophoretic isolation, the L. albus MMP-9 inhibitor was found to comprise a large fragment from δ-conglutin and, to a lower extent, small fragments of β-conglutin. In vitro studies showed that the MMPI successfully inhibited MMP-9 activity in a dose-dependent manner in colon cancer cells, with an IC50 of 10 µg·mL-1 without impairing gene expression nor cell growth. In vivo studies showed that the MMPI maintained its bioactivities when administered orally and significantly reduced colitis symptoms, along with a very significant inhibition of MMP-2 and -9 activities. Overall, results reveal a novel type of MMPI in lupine that is edible, proteinaceous in nature and soluble in water, and effective in vivo, suggesting a high potential application as a nutraceutical or a functional food in pathologies related to abnormally high MMP-9 activity in the digestive system.

• Authors: Joana Mota, Rosa Direito, João Rocha, João Fernandes, Bruno Sepodes, Maria Eduardo Figueira, Anabela Raymundo, Ana Lima, Ricardo Boavida Ferreira
• Journal: International journal of molecular sciences (Int J Mol Sci) Vol. 22 Issue 24 (Dec 10 2021) ISSN: 1422-0067 [Electronic] Switzerland
• PMID: 34948082 (Publication Type: Journal Article)

Chemical References

• Matrix Metalloproteinase Inhibitors
• Plant Proteins
• Matrix Metalloproteinase 2
• Matrix Metalloproteinase 9

Topics

• Animals
• Colitis (diet therapy, drug therapy, enzymology)
• Female
• HT29 Cells
• Humans
• Lupinus (chemistry, metabolism)
• Male
• Matrix Metalloproteinase 2 (drug effects, metabolism)
• Matrix Metalloproteinase 9 (drug effects, metabolism)
• Matrix Metalloproteinase Inhibitors (isolation & purification, pharmacology)
• Mice
• Plant Proteins (isolation & purification, pharmacology)
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Tandem Mass Spectrometry