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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| Format: | Thesis |
| Language: | English |
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2015
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| Online Access: | https://ir.uitm.edu.my/id/eprint/15672/1/TM_NUR%20HIDAYAH%20MOHD%20ISHAK%20MD%2015_5.pdf |
| _version_ | 1796900258409611264 |
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| author | Mohd Ishak, Nur Hidayah |
| author_facet | Mohd Ishak, Nur Hidayah |
| author_sort | Mohd Ishak, Nur Hidayah |
| collection | UITM |
| 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. |
| first_indexed | 2024-03-06T01:33:00Z |
| format | Thesis |
| id | oai:ir.uitm.edu.my:15672 |
| institution | Universiti Teknologi MARA |
| language | English |
| last_indexed | 2024-03-06T01:33:00Z |
| publishDate | 2015 |
| record_format | dspace |
| spelling | oai:ir.uitm.edu.my: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 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/15672/1/TM_NUR%20HIDAYAH%20MOHD%20ISHAK%20MD%2015_5.pdf |
| work_keys_str_mv | AT mohdishaknurhidayah effectsoftheaflavinsrichfractiononadhesionmoleculesandinflammationvianfkbpathwayinstimulatedendothelialcellsnurhidayahmohdishak |