Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir

Matrix metalloproteinases (MMPs) can regulate extracellular matrix (ECM) turnover in the trabecular meshwork (TM) of eye. Reduced MMP secretion increases ECM deposition in TM, which is associated with increased intraocular pressure (IOP). Increased IOP is the major risk factor for glaucoma, the lead...

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Main Author:	Mohd Nasir, Normie Aida
Format:	Thesis
Language:	English
Published:	2019
Subjects:	Cytology Organic substances
Online Access:	https://ir.uitm.edu.my/id/eprint/30145/1/30145.pdf
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spelling	my-uitm-ir.301452022-07-20T00:01:31Z Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir 2019-07 Mohd Nasir, Normie Aida Cytology Organic substances Matrix metalloproteinases (MMPs) can regulate extracellular matrix (ECM) turnover in the trabecular meshwork (TM) of eye. Reduced MMP secretion increases ECM deposition in TM, which is associated with increased intraocular pressure (IOP). Increased IOP is the major risk factor for glaucoma, the leading cause of irreversible blindness, worldwide. Previously trans-resveratrol was shown to reduce IOP in normotensive and oculohypertensive rats. The dose- and time-dependent effects of trans-resveratrol on human TM cells (HTMCs) viability, MMP-2 and -9 secretion and the mechanisms involved remain unknown and hence these effects were investigated in the present study. HTMCs were cultured and divided into 11 groups that received treatment with DMSO (0.1%), dexamethasone (100 nM), and trans-resveratrol (3.125, 6.25, 12.5, 25 and 50 µM) either in the presence or absence of dexamethasone for 2, 5 and 7 days. To study the role of A₁ adenosine receptors (A₁AR) and nuclear factor kappa B (NFĸB) in trans-resveratrol mediated effects on MMP secretions, additional groups of HTMCs were pre-treated with dipropylcyclopentylxanthine (DPCPX), a specific A₁AR antagonist and curcumin, an inhibitor of NFĸB. Cell viability was determined by using MTS assay while MMP-2 and -9 expressions were investigated using western blot. A₁AR expression was determined using western blot and ELISA while NFĸB activation was determined using immunocytochemistry and ELISA. This study demonstrated that incubation of HTMCs with trans-resveratrol up to a concentration of 25 μM does not affect the viability but at 50 µM, it significantly reduces viability of HTMCs both in the presence and absence of dexamethasone. This effect of trans-resveratrol on the viability of HTMCs was dose-dependent but not time-dependent. Dexamethasone reduced MMP-2 and -9 expressions after 5 and 7 days treatment, compared to untreated group (p<0.01). Incubation with 12.5 µM trans-resveratrol for 5 days in the presence of dexamethasone increased MMP-2 and 9 level compared to dexamethasone-treated group (p<0.05). Significant reduction of A₁AR expression was seen in dexamethasone-treated group compared to untreated group (p<0.01). Trans-resveratrol significantly increased A₁AR expression compared to dexamethasone treated and DPCPX-treated groups (p<0.001). Dexamethasone significantly reduced NFĸB activation compared to the untreated group (p<0.001). Increased nuclear localization of phospho p65 NFĸB was seen in trans-resveratrol treated group in the presence and absence of dexamethasone. In conclusion, transresveratrol counteracts the effects of dexamethasone on MMP-2 and -9 levels of HTMCs, which could be mediated by A₁AR and NFĸB. 2019-07 Thesis https://ir.uitm.edu.my/id/eprint/30145/ https://ir.uitm.edu.my/id/eprint/30145/1/30145.pdf text en public masters Universiti Teknologi MARA (Kampus Sg. Buloh) Faculty of Medicine Agarwal, Renu Krasilnikova, Anna Sheikh Abdul Kadir, Siti Hamimah
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Agarwal, Renu Krasilnikova, Anna Sheikh Abdul Kadir, Siti Hamimah
topic	Cytology Organic substances
spellingShingle	Cytology Organic substances Mohd Nasir, Normie Aida Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir
description	Matrix metalloproteinases (MMPs) can regulate extracellular matrix (ECM) turnover in the trabecular meshwork (TM) of eye. Reduced MMP secretion increases ECM deposition in TM, which is associated with increased intraocular pressure (IOP). Increased IOP is the major risk factor for glaucoma, the leading cause of irreversible blindness, worldwide. Previously trans-resveratrol was shown to reduce IOP in normotensive and oculohypertensive rats. The dose- and time-dependent effects of trans-resveratrol on human TM cells (HTMCs) viability, MMP-2 and -9 secretion and the mechanisms involved remain unknown and hence these effects were investigated in the present study. HTMCs were cultured and divided into 11 groups that received treatment with DMSO (0.1%), dexamethasone (100 nM), and trans-resveratrol (3.125, 6.25, 12.5, 25 and 50 µM) either in the presence or absence of dexamethasone for 2, 5 and 7 days. To study the role of A₁ adenosine receptors (A₁AR) and nuclear factor kappa B (NFĸB) in trans-resveratrol mediated effects on MMP secretions, additional groups of HTMCs were pre-treated with dipropylcyclopentylxanthine (DPCPX), a specific A₁AR antagonist and curcumin, an inhibitor of NFĸB. Cell viability was determined by using MTS assay while MMP-2 and -9 expressions were investigated using western blot. A₁AR expression was determined using western blot and ELISA while NFĸB activation was determined using immunocytochemistry and ELISA. This study demonstrated that incubation of HTMCs with trans-resveratrol up to a concentration of 25 μM does not affect the viability but at 50 µM, it significantly reduces viability of HTMCs both in the presence and absence of dexamethasone. This effect of trans-resveratrol on the viability of HTMCs was dose-dependent but not time-dependent. Dexamethasone reduced MMP-2 and -9 expressions after 5 and 7 days treatment, compared to untreated group (p<0.01). Incubation with 12.5 µM trans-resveratrol for 5 days in the presence of dexamethasone increased MMP-2 and 9 level compared to dexamethasone-treated group (p<0.05). Significant reduction of A₁AR expression was seen in dexamethasone-treated group compared to untreated group (p<0.01). Trans-resveratrol significantly increased A₁AR expression compared to dexamethasone treated and DPCPX-treated groups (p<0.001). Dexamethasone significantly reduced NFĸB activation compared to the untreated group (p<0.001). Increased nuclear localization of phospho p65 NFĸB was seen in trans-resveratrol treated group in the presence and absence of dexamethasone. In conclusion, transresveratrol counteracts the effects of dexamethasone on MMP-2 and -9 levels of HTMCs, which could be mediated by A₁AR and NFĸB.
format	Thesis
qualification_level	Master's degree
author	Mohd Nasir, Normie Aida
author_facet	Mohd Nasir, Normie Aida
author_sort	Mohd Nasir, Normie Aida
title	Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir
title_short	Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir
title_full	Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir
title_fullStr	Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir
title_full_unstemmed	Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir
title_sort	protective effect of trans-resveratrol on dexamethasone-induced changes in matrix metalloproteinases expression by human trabecular meshwork cells: potential mechanisms / normie aida mohd nasir
granting_institution	Universiti Teknologi MARA (Kampus Sg. Buloh)
granting_department	Faculty of Medicine
publishDate	2019
url	https://ir.uitm.edu.my/id/eprint/30145/1/30145.pdf
_version_	1783734085460426752

Protective effect of Trans-resveratrol on Dexamethasone-induced changes in Matrix Metalloproteinases Expression by Human Trabecular Meshwork Cells: potential mechanisms / Normie Aida Mohd Nasir

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