Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak

Theaflavins is the main polyphenolic compounds in black tea that suggested to have anti-inflammation mechanism. This study aim to determine the effects of theaflavinsrich fraction (TsRF) in Lipopolysaccharide(LPS)- stimulated Human Umbilical Vein Endothelial cells (HUVECs) on cellular adhesion molec...

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Main Author: Mohd Ishak, Nur Hidayah
Format: Thesis
Language:English
Published: 2015
Subjects:
Online Access:https://ir.uitm.edu.my/id/eprint/15672/
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author Mohd Ishak, Nur Hidayah
author_facet Mohd Ishak, Nur Hidayah
author_sort Mohd Ishak, Nur Hidayah
building UiTM Institutional Repository
collection Online Access
description Theaflavins is the main polyphenolic compounds in black tea that suggested to have anti-inflammation mechanism. This study aim to determine the effects of theaflavinsrich fraction (TsRF) in Lipopolysaccharide(LPS)- stimulated Human Umbilical Vein Endothelial cells (HUVECs) on cellular adhesion molecules (CAMs; E-selectin, vascular cellular adhesion molecules;VCAM-1 and intercellular adhesion molecule; ICAM-1) and Nuclear Faktor-kappa E^NF-tcB; protein subunits p50 and p65) in both expression in gene and protein expression level. Cytotoxicity was assessed by methylthiazol- tetrazolium(MTT) assay using TsRF(Organics Herbs, China) concentrations ranging from 1.6 to 200 /xg/ml which were added to HUVECs (Cascade Biologies,USA). Confluent HUVECs were treated with LPS (Sigma,USA) and TsRF. After incubation of 16 hours, protein expression of E-selectin, VCAM-1, ICAM-1, NF- kB p50 and p65 were measured by Enzyme-linked immunosorbent assay (ELISA). Total RNA was isolated from cell pellets using the RNeasy kit (Qiagenlnc, Valencia,CA) and Gene expressions were determined by quantitative Real-Time Polymerase Chain Reaction (qPCR). TsRF <50 |xg/mL seduced viability cell to > 80%. TsRF effectively inhibited LPS-stimulated expression of E-selectin, VCAM-1 and ICAM-1 in HUVECs at the transcription level with TsRF 10-50 (ig/mL reduced of E-selectin (p<0.0001), VCAM-1 (p<0.0001), and ICAM-1 (p<0.0001). While in translation level, TsRF 10-50 |ig/mL reduced E-selectin (p<0.0001), VCAM-1 (p<0.0001), and ICAM-1 (p<0.05). Suppression of protein subunit p50 and p65 indicate that TsRF blocked the activation of NF-tcB mechanism. In gene level, TsRF 10-50 |ig/mL reduced protein subunits p65 (0.0001) and p50 (p<0.05).While expression of NF-tcB p65 (p<0.01) shown inhibition at 50 ug/ml in protein level.These results suggest that TsRF has anti-inflammatory mechanism that involves by inhibit cellular adhesion molecules expression and blocking the NF-tcB activation thus suggesting potential benefits in preventing atherosclerosis.
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spelling uitm-156722022-03-09T08:38:01Z https://ir.uitm.edu.my/id/eprint/15672/ Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak Mohd Ishak, Nur Hidayah Drugs and their actions Materia medica Theaflavins is the main polyphenolic compounds in black tea that suggested to have anti-inflammation mechanism. This study aim to determine the effects of theaflavinsrich fraction (TsRF) in Lipopolysaccharide(LPS)- stimulated Human Umbilical Vein Endothelial cells (HUVECs) on cellular adhesion molecules (CAMs; E-selectin, vascular cellular adhesion molecules;VCAM-1 and intercellular adhesion molecule; ICAM-1) and Nuclear Faktor-kappa E^NF-tcB; protein subunits p50 and p65) in both expression in gene and protein expression level. Cytotoxicity was assessed by methylthiazol- tetrazolium(MTT) assay using TsRF(Organics Herbs, China) concentrations ranging from 1.6 to 200 /xg/ml which were added to HUVECs (Cascade Biologies,USA). Confluent HUVECs were treated with LPS (Sigma,USA) and TsRF. After incubation of 16 hours, protein expression of E-selectin, VCAM-1, ICAM-1, NF- kB p50 and p65 were measured by Enzyme-linked immunosorbent assay (ELISA). Total RNA was isolated from cell pellets using the RNeasy kit (Qiagenlnc, Valencia,CA) and Gene expressions were determined by quantitative Real-Time Polymerase Chain Reaction (qPCR). TsRF <50 |xg/mL seduced viability cell to > 80%. TsRF effectively inhibited LPS-stimulated expression of E-selectin, VCAM-1 and ICAM-1 in HUVECs at the transcription level with TsRF 10-50 (ig/mL reduced of E-selectin (p<0.0001), VCAM-1 (p<0.0001), and ICAM-1 (p<0.0001). While in translation level, TsRF 10-50 |ig/mL reduced E-selectin (p<0.0001), VCAM-1 (p<0.0001), and ICAM-1 (p<0.05). Suppression of protein subunit p50 and p65 indicate that TsRF blocked the activation of NF-tcB mechanism. In gene level, TsRF 10-50 |ig/mL reduced protein subunits p65 (0.0001) and p50 (p<0.05).While expression of NF-tcB p65 (p<0.01) shown inhibition at 50 ug/ml in protein level.These results suggest that TsRF has anti-inflammatory mechanism that involves by inhibit cellular adhesion molecules expression and blocking the NF-tcB activation thus suggesting potential benefits in preventing atherosclerosis. 2015 Thesis NonPeerReviewed text en https://ir.uitm.edu.my/id/eprint/15672/1/TM_NUR%20HIDAYAH%20MOHD%20ISHAK%20MD%2015_5.pdf Mohd Ishak, Nur Hidayah (2015) Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak. (2015) Masters thesis, thesis, Universiti Teknologi MARA. <http://terminalib.uitm.edu.my/15672.pdf>
spellingShingle Drugs and their actions
Materia medica
Mohd Ishak, Nur Hidayah
Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak
title Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak
title_full Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak
title_fullStr Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak
title_full_unstemmed Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak
title_short Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak
title_sort effects of theaflavins-rich fraction on adhesion molecules and inflammation via nf-kb pathway in stimulated endothelial cells / nur hidayah mohd ishak
topic Drugs and their actions
Materia medica
url https://ir.uitm.edu.my/id/eprint/15672/