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

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:
Drugs and their actions
Materia medica
Online Access:https://ir.uitm.edu.my/id/eprint/15672/1/TM_NUR%20HIDAYAH%20MOHD%20ISHAK%20MD%2015_5.pdf
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spelling my-uitm-ir.156722022-03-09T08:38:01Z Effects of Theaflavins-rich fraction on adhesion molecules and inflammation via NF-KB pathway in stimulated Endothelial cells / Nur Hidayah Mohd Ishak 2015 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 https://ir.uitm.edu.my/id/eprint/15672/ https://ir.uitm.edu.my/id/eprint/15672/1/TM_NUR%20HIDAYAH%20MOHD%20ISHAK%20MD%2015_5.pdf text en public mphil masters Universiti Teknologi MARA Faculty of Medicine
institution Universiti Teknologi MARA
collection UiTM Institutional Repository
language English
topic Drugs and their actions
Materia medica
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
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.
format Thesis
qualification_name Master of Philosophy (M.Phil.)
qualification_level Master's degree
author Mohd Ishak, Nur Hidayah
author_facet Mohd Ishak, Nur Hidayah
author_sort Mohd Ishak, Nur Hidayah
title 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_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_sort effects of theaflavins-rich fraction on adhesion molecules and inflammation via nf-kb pathway in stimulated endothelial cells / nur hidayah mohd ishak
granting_institution Universiti Teknologi MARA
granting_department Faculty of Medicine
publishDate 2015
url https://ir.uitm.edu.my/id/eprint/15672/1/TM_NUR%20HIDAYAH%20MOHD%20ISHAK%20MD%2015_5.pdf
_version_ 1783733440894468096

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