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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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Cresyl violet and TUNEL staining in the CA3 region of OHSC. Cresyl violet staining was performed at the end of 48 h recovery after KA treatment for 15 h. Representative 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 and C). Cresyl violet-positive cells regarded as survival cells and TUNEL-positive cells indicated as apoptotic cell death were quantified in CA3 region (B and D). Data are presented as means ± S.E.M. of 6 experiments for cresyl violet and 5 experiments for TUNEL staining. Asterisks (*) indicate statistically significant difference from control (* p < 0.001 in (B) and * p < 0.001 in (D)); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.
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f2-ijms-14-18256: Cresyl violet and TUNEL staining in the CA3 region of OHSC. Cresyl violet staining was performed at the end of 48 h recovery after KA treatment for 15 h. Representative 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 and C). Cresyl violet-positive cells regarded as survival cells and TUNEL-positive cells indicated as apoptotic cell death were quantified in CA3 region (B and D). Data are presented as means ± S.E.M. of 6 experiments for cresyl violet and 5 experiments for TUNEL staining. Asterisks (*) indicate statistically significant difference from control (* p < 0.001 in (B) and * p < 0.001 in (D)); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.

Mentions: Slices were stained with cresyl violet in order to measure cell survival and to determine the agreement with results from PI and a terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL) staining. As shown in Figure 2A, the cresyl violet stained sections showed the KA-induced neuronal death was concentrated in the CA3 pyramidal neurons. However, cell death was significantly prevented by treatment with ATTP or ATTN (100 μM) compared to treatment with KA alone, although there were no statistically significant differences between ATTP and ATTN treatment for either co-treatment or post-treatment (Figure 2B).


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)

Cresyl violet and TUNEL staining in the CA3 region of OHSC. Cresyl violet staining was performed at the end of 48 h recovery after KA treatment for 15 h. Representative 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 and C). Cresyl violet-positive cells regarded as survival cells and TUNEL-positive cells indicated as apoptotic cell death were quantified in CA3 region (B and D). Data are presented as means ± S.E.M. of 6 experiments for cresyl violet and 5 experiments for TUNEL staining. Asterisks (*) indicate statistically significant difference from control (* p < 0.001 in (B) and * p < 0.001 in (D)); 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

f2-ijms-14-18256: Cresyl violet and TUNEL staining in the CA3 region of OHSC. Cresyl violet staining was performed at the end of 48 h recovery after KA treatment for 15 h. Representative 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 and C). Cresyl violet-positive cells regarded as survival cells and TUNEL-positive cells indicated as apoptotic cell death were quantified in CA3 region (B and D). Data are presented as means ± S.E.M. of 6 experiments for cresyl violet and 5 experiments for TUNEL staining. Asterisks (*) indicate statistically significant difference from control (* p < 0.001 in (B) and * p < 0.001 in (D)); Sharps (#) indicate statistically significant difference from KA-treated cultures (#p < 0.05). Scale bar: 200 μm.
Mentions: Slices were stained with cresyl violet in order to measure cell survival and to determine the agreement with results from PI and a terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling (TUNEL) staining. As shown in Figure 2A, the cresyl violet stained sections showed the KA-induced neuronal death was concentrated in the CA3 pyramidal neurons. However, cell death was significantly prevented by treatment with ATTP or ATTN (100 μM) compared to treatment with KA alone, although there were no statistically significant differences between ATTP and ATTN treatment for either co-treatment or post-treatment (Figure 2B).

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