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Neuroprotective effects of α-tocotrienol on kainic acid-induced neurotoxicity in organotypic hippocampal slice cultures.

Jung NY, Lee KH, Won R, Lee BH - Int J Mol Sci (2013)

Bottom Line: Both co-treatment and post-treatment of ATPH (100 µM) or ATTN (100 µM) significantly increased the cell survival and reduced the number of TUNEL-positive cells in the CA3 region.These data suggest that ATPH and ATTN treatment have protective effects on KA-induced cell death in OHSC.ATTN treatment tended to be more effective than ATPH treatment, even though there was no significant difference between ATPH and ATTN in co-treatment or post-treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, Korea. bhlee@yuhs.ac.

ABSTRACT
Vitamin E, such as alpha-tocopherol (ATPH) and alpha-tocotrienol (ATTN), is a chain-breaking antioxidant that prevents the chain propagation step during lipid peroxidation. In the present study, we investigated the effects of ATTN on KA-induced neuronal death using organotypic hippocampal slice culture (OHSC) and compared the neuroprotective effects of ATTN and ATPH. After 15 h KA (5 µM) treatment, delayed neuronal death was detected in the CA3 region and reactive oxygen species (ROS) formation and lipid peroxidation were also increased. Both co-treatment and post-treatment of ATPH (100 µM) or ATTN (100 µM) significantly increased the cell survival and reduced the number of TUNEL-positive cells in the CA3 region. Increased dichlorofluorescein (DCF) fluorescence and levels of thiobarbiturate reactive substances (TBARS) were decreased by ATPH and ATTN treatment. These data suggest that ATPH and ATTN treatment have protective effects on KA-induced cell death in OHSC. ATTN treatment tended to be more effective than ATPH treatment, even though there was no significant difference between ATPH and ATTN in co-treatment or post-treatment.

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Effects of alpha-tocopherol (ATPH) and alpha-tocotrienol (ATTN) on KA-induced PI uptake in organotypic hippocampal slice culture (OHSC). Representative PI uptake images from CONT (untreated), KA (5 μM KA only treated), KA/ATPH Co-treat (100 μM ATPH with KA), KA/ATPH Post-treat (100 μM ATPH after KA), KA/ATTN Co-treat (100 μM ATTN with KA), and KA/ATTN Post-treat (100 μM ATTN after KA) slices (A). Quantification of PI images. Data are presented as means ± S.E.M. of 5 to 7 experiments (B). Asterisks (*) indicate statistically significant difference from control (* p < 0.001); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.
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f1-ijms-14-18256: Effects of alpha-tocopherol (ATPH) and alpha-tocotrienol (ATTN) on KA-induced PI uptake in organotypic hippocampal slice culture (OHSC). Representative PI uptake images from CONT (untreated), KA (5 μM KA only treated), KA/ATPH Co-treat (100 μM ATPH with KA), KA/ATPH Post-treat (100 μM ATPH after KA), KA/ATTN Co-treat (100 μM ATTN with KA), and KA/ATTN Post-treat (100 μM ATTN after KA) slices (A). Quantification of PI images. Data are presented as means ± S.E.M. of 5 to 7 experiments (B). Asterisks (*) indicate statistically significant difference from control (* p < 0.001); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.

Mentions: PI uptake was measured to assess the effects of ATPH and ATTN on KA-induced cell death. When hippocampal slices were exposed to 5 μM KA for 15 h, the PI uptake in the CA3 region was significantly higher than that in the CA1 region (Figure 1A). Treatment with ATPH significantly reduced the PI uptake at 48 h after treatment as compared to post-treatment with ATPH. Treatment with ATTN also showed significantly reduced PI uptake in both co- and post- treatment at 48 h after treatment. Thus, co-treatment using ATPH (100 μM) or ATTN (100 μM) with KA significantly reduced the level of PI uptake in the CA3 region compared with KA treatment alone (Figure 1B). Further, the same trend was also observed with post-treatment using ATPH or ATTN 15 h after KA treatment.


Neuroprotective effects of α-tocotrienol on kainic acid-induced neurotoxicity in organotypic hippocampal slice cultures.

Jung NY, Lee KH, Won R, Lee BH - Int J Mol Sci (2013)

Effects of alpha-tocopherol (ATPH) and alpha-tocotrienol (ATTN) on KA-induced PI uptake in organotypic hippocampal slice culture (OHSC). Representative PI uptake images from CONT (untreated), KA (5 μM KA only treated), KA/ATPH Co-treat (100 μM ATPH with KA), KA/ATPH Post-treat (100 μM ATPH after KA), KA/ATTN Co-treat (100 μM ATTN with KA), and KA/ATTN Post-treat (100 μM ATTN after KA) slices (A). Quantification of PI images. Data are presented as means ± S.E.M. of 5 to 7 experiments (B). Asterisks (*) indicate statistically significant difference from control (* p < 0.001); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3794779&req=5

f1-ijms-14-18256: Effects of alpha-tocopherol (ATPH) and alpha-tocotrienol (ATTN) on KA-induced PI uptake in organotypic hippocampal slice culture (OHSC). Representative PI uptake images from CONT (untreated), KA (5 μM KA only treated), KA/ATPH Co-treat (100 μM ATPH with KA), KA/ATPH Post-treat (100 μM ATPH after KA), KA/ATTN Co-treat (100 μM ATTN with KA), and KA/ATTN Post-treat (100 μM ATTN after KA) slices (A). Quantification of PI images. Data are presented as means ± S.E.M. of 5 to 7 experiments (B). Asterisks (*) indicate statistically significant difference from control (* p < 0.001); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.
Mentions: PI uptake was measured to assess the effects of ATPH and ATTN on KA-induced cell death. When hippocampal slices were exposed to 5 μM KA for 15 h, the PI uptake in the CA3 region was significantly higher than that in the CA1 region (Figure 1A). Treatment with ATPH significantly reduced the PI uptake at 48 h after treatment as compared to post-treatment with ATPH. Treatment with ATTN also showed significantly reduced PI uptake in both co- and post- treatment at 48 h after treatment. Thus, co-treatment using ATPH (100 μM) or ATTN (100 μM) with KA significantly reduced the level of PI uptake in the CA3 region compared with KA treatment alone (Figure 1B). Further, the same trend was also observed with post-treatment using ATPH or ATTN 15 h after KA treatment.

Bottom Line: Both co-treatment and post-treatment of ATPH (100 µM) or ATTN (100 µM) significantly increased the cell survival and reduced the number of TUNEL-positive cells in the CA3 region.These data suggest that ATPH and ATTN treatment have protective effects on KA-induced cell death in OHSC.ATTN treatment tended to be more effective than ATPH treatment, even though there was no significant difference between ATPH and ATTN in co-treatment or post-treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, Seoul 120-752, Korea. bhlee@yuhs.ac.

ABSTRACT
Vitamin E, such as alpha-tocopherol (ATPH) and alpha-tocotrienol (ATTN), is a chain-breaking antioxidant that prevents the chain propagation step during lipid peroxidation. In the present study, we investigated the effects of ATTN on KA-induced neuronal death using organotypic hippocampal slice culture (OHSC) and compared the neuroprotective effects of ATTN and ATPH. After 15 h KA (5 µM) treatment, delayed neuronal death was detected in the CA3 region and reactive oxygen species (ROS) formation and lipid peroxidation were also increased. Both co-treatment and post-treatment of ATPH (100 µM) or ATTN (100 µM) significantly increased the cell survival and reduced the number of TUNEL-positive cells in the CA3 region. Increased dichlorofluorescein (DCF) fluorescence and levels of thiobarbiturate reactive substances (TBARS) were decreased by ATPH and ATTN treatment. These data suggest that ATPH and ATTN treatment have protective effects on KA-induced cell death in OHSC. ATTN treatment tended to be more effective than ATPH treatment, even though there was no significant difference between ATPH and ATTN in co-treatment or post-treatment.

Show MeSH