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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.50 [2012] >
IJEB Vol.50(08) [August 2012] >
| Title: | Radiomodulation by Hoechst 33258 against radiation- induced damage in murine splenocytes |
| Authors: | Khatoon, Zubaida
Kale, R K |
| Keywords: | Antioxidant enzymes
FACS
Hoechst 33258
Membrane fluidity
Peroxidative damage
Radioprotector |
| Issue Date: | Aug-2012 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | In this study modulatory effect of Hoechst 33258 on radiation induced
membrane related signaling events which ultimately leads to apoptosis has been
investigated. Splenocytes from swiss albino mice were irradiated in air at room
temperature in a gamma chamber (240 TBq 60Co Model 4000 A) at the
dose-rate of 0.052 Gys-1. Membrane lipid peroxidation, fluidity, specific
activities of antioxidant enzymes, levels of nitric oxide, glutathione and
apoptosis in presence and absence of different concentrations of Hoechst 33258
has been assayed. DNA binding activity of nuclear factor kappa
B and activator protein–1 was
also assayed by
electrophoretic mobility shift assay. Modulatory effect of Hoechst
33258 was examined at 3 and 5 Gy using different concentrations (10, 20 and 30 µM). Hoechst 33258 was found
to inhibit radiation induced peroxidative damage and fluidity and lowered the
level of nitric
oxide and
apoptosis - as evident by DNA ladder assay and FACS, indicating free radicals
scavenging potential. Dot plot
diagramme clearly showed that 30 µM Hoechst 33258 caused 14% and 19%
decrease in apoptotic cells at 3 Gy and 5 Gy of radiation respectively
(compared to irradiated control group). Further DNA binding activity of
nuclear factor kappa B and activator protein–1 was also inhibited but the
antioxidant potential of the cells was enhanced. These findings support that Hoechst 33258
protects the cell from undergoing apoptosis. Hoechst 33258 may have interacted
and has an ability to protect splenocytes against radiation induced apoptosis
through modulation of membrane-related signaling events and antioxidant status. |
| Page(s): | 517-530 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.50(08) [August 2012]
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