The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib

Glutamate excitotoxicity is a concept that underpinned the massive release of glutamate to extracellular space, thereby inducing neuronal and glial injuries. Hypothermia has been proposed to offer neuroprotection even though the mechanism underlying it is poorly understood. Furthermore, prolonged ti...

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Main Author:	Abd Talib, Fatin Nur Asyiqin
Format:	Thesis
Language:	English
Published:	2018
Subjects:	Organic substances
Online Access:	https://ir.uitm.edu.my/id/eprint/63438/1/63438.pdf
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id	my-uitm-ir.63438
record_format	uketd_dc
spelling	my-uitm-ir.634382022-07-20T00:01:19Z The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib 2018-05 Abd Talib, Fatin Nur Asyiqin Organic substances Medical emergencies. Critical care. Intensive care. First aid Glutamate excitotoxicity is a concept that underpinned the massive release of glutamate to extracellular space, thereby inducing neuronal and glial injuries. Hypothermia has been proposed to offer neuroprotection even though the mechanism underlying it is poorly understood. Furthermore, prolonged time is required for hypothermia to exert its effects as well as inconsistencies and variation in the outcomes patients. An adjuvant therapy with hypothermia may be an alternative to reduce exposure time and obtain consistent outcomes. Progesterone (P4) is a neurosteroid which has been shown to elicit neuroprotection in neuronal cells with ischemic injury. This study investigates the neuroprotective effects of hypothermia and P4 on astrocytes following glutamateinduced toxicity. The cultured primary cortical astrocyte cells were exposed to 50 µM of glutamate for 15 minutes followed by incubation under hypothermia conditions with and without P4 for 24 hours. After 24 hours, the viability of cells was assessed by 3[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The intracellular concentration of glutamate in astrocyte cells was estimated by the glutamate uptake assay. The levels of p62 and S100β were measured using ELISA. The membrane protein was extracted and estimated for GLT-1 by Western Blot. There were significant increases in the percentage of viable cells as well as the concentration of glutamate uptake by the astrocyte cells in mild hypothermia with P4 (88.27 ± 3.96%) and moderate hypothermia with P4 (80.94 ± 4.12%) as compared to normothermia after glutamate induced toxicity (51.05 ± 4.10%). Further analysis revealed that there was a significant effect of moderate hypothermia with P4 (142.78 ± 13.85 pg/ml) in increasing the S100β level in comparison to normothermia across the glutamate-induced toxicity groups (74.34 ± 4.42 pg/ml) and significantly increasing in glutamate uptake (p ≤ 0.01) after treated with both mild and moderate hypothermia and P4. There was a significant increase of membrane GLT-1 in both mild hypothermia (1.07 ± 0.01 FD) and moderate hypothermia (0.87 ± 0.08 FD) group when compared to the normothermia (0.53 ± 0.05 FD) group. The p62 level was shown significant reduced in both mild and moderate hypothermia and P4 (p ≤ 0.01) in comparison to normothermia across the glutamateinduced toxicity group. In conclusion, hypothermia and P4 reduced the glutamateinduced toxicity in the astrocyte cells by increasing glutamate uptake via GLT-1. 2018-05 Thesis https://ir.uitm.edu.my/id/eprint/63438/ https://ir.uitm.edu.my/id/eprint/63438/1/63438.pdf text en public masters Universiti Teknologi MARA (Kampus Sg. Buloh) Faculty of Medicine Siran, Rosfaiizah Mat Zain, Mazatulikhma Abd Aziz, Noor Azean Anis Husin, Andrean
institution	Universiti Teknologi MARA
collection	UiTM Institutional Repository
language	English
advisor	Siran, Rosfaiizah Mat Zain, Mazatulikhma Abd Aziz, Noor Azean Anis Husin, Andrean
topic	Organic substances Organic substances
spellingShingle	Organic substances Organic substances Abd Talib, Fatin Nur Asyiqin The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib
description	Glutamate excitotoxicity is a concept that underpinned the massive release of glutamate to extracellular space, thereby inducing neuronal and glial injuries. Hypothermia has been proposed to offer neuroprotection even though the mechanism underlying it is poorly understood. Furthermore, prolonged time is required for hypothermia to exert its effects as well as inconsistencies and variation in the outcomes patients. An adjuvant therapy with hypothermia may be an alternative to reduce exposure time and obtain consistent outcomes. Progesterone (P4) is a neurosteroid which has been shown to elicit neuroprotection in neuronal cells with ischemic injury. This study investigates the neuroprotective effects of hypothermia and P4 on astrocytes following glutamateinduced toxicity. The cultured primary cortical astrocyte cells were exposed to 50 µM of glutamate for 15 minutes followed by incubation under hypothermia conditions with and without P4 for 24 hours. After 24 hours, the viability of cells was assessed by 3[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. The intracellular concentration of glutamate in astrocyte cells was estimated by the glutamate uptake assay. The levels of p62 and S100β were measured using ELISA. The membrane protein was extracted and estimated for GLT-1 by Western Blot. There were significant increases in the percentage of viable cells as well as the concentration of glutamate uptake by the astrocyte cells in mild hypothermia with P4 (88.27 ± 3.96%) and moderate hypothermia with P4 (80.94 ± 4.12%) as compared to normothermia after glutamate induced toxicity (51.05 ± 4.10%). Further analysis revealed that there was a significant effect of moderate hypothermia with P4 (142.78 ± 13.85 pg/ml) in increasing the S100β level in comparison to normothermia across the glutamate-induced toxicity groups (74.34 ± 4.42 pg/ml) and significantly increasing in glutamate uptake (p ≤ 0.01) after treated with both mild and moderate hypothermia and P4. There was a significant increase of membrane GLT-1 in both mild hypothermia (1.07 ± 0.01 FD) and moderate hypothermia (0.87 ± 0.08 FD) group when compared to the normothermia (0.53 ± 0.05 FD) group. The p62 level was shown significant reduced in both mild and moderate hypothermia and P4 (p ≤ 0.01) in comparison to normothermia across the glutamateinduced toxicity group. In conclusion, hypothermia and P4 reduced the glutamateinduced toxicity in the astrocyte cells by increasing glutamate uptake via GLT-1.
format	Thesis
qualification_level	Master's degree
author	Abd Talib, Fatin Nur Asyiqin
author_facet	Abd Talib, Fatin Nur Asyiqin
author_sort	Abd Talib, Fatin Nur Asyiqin
title	The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib
title_short	The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib
title_full	The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib
title_fullStr	The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib
title_full_unstemmed	The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib
title_sort	effect of hypothermia and progesterone (p4) against glutamate challenged primary cortical astrocytes on s100β, glutamate uptake, glt-1 and p62 / fatin nur asyiqin abd talib
granting_institution	Universiti Teknologi MARA (Kampus Sg. Buloh)
granting_department	Faculty of Medicine
publishDate	2018
url	https://ir.uitm.edu.my/id/eprint/63438/1/63438.pdf
_version_	1783735323502575616

The effect of Hypothermia and Progesterone (P4) against Glutamate challenged primary cortical astrocytes on S100β, Glutamate Uptake, GLT-1 and p62 / Fatin Nur Asyiqin Abd Talib

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