Please use this identifier to cite or link to this item: http://nopr.niscair.res.in/handle/123456789/14482
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: <span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">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 <sup>60</sup>Co Model 4000 A) at the dose-rate of 0.052 Gys<sup>-1</sup>.<span style="font-size: 11.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-bidi-font-family:mangal;mso-ansi-language:en-sg;mso-fareast-language:en-sg;="" mso-bidi-language:hi"="" lang="EN-SG"> 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. <span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">DNA binding<span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-sg;mso-fareast-language:en-sg;mso-bidi-language:="" hi"="" lang="EN-SG"> activity of <span style="font-size:11.0pt;font-family: " times="" new="" roman";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">nuclear factor kappa B<span style="font-size:11.0pt;font-family:" times="" new="" roman";="" mso-fareast-font-family:"times="" roman";mso-bidi-font-family:mangal;="" mso-ansi-language:en-sg;mso-fareast-language:en-sg;mso-bidi-language:hi"="" lang="EN-GB"> <span style="font-size:11.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:="" "times="" roman";mso-bidi-font-family:mangal;mso-ansi-language:en-gb;="" mso-fareast-language:en-us;mso-bidi-language:hi"="" lang="EN-GB">and activator protein–1 was also assayed <span style="font-size:11.0pt;font-family:" times="" new="" roman";="" mso-fareast-font-family:"times="" roman";mso-bidi-font-family:mangal;="" mso-ansi-language:en-sg;mso-fareast-language:en-sg;mso-bidi-language:hi"="" lang="EN-SG">by electrophoretic mobility shift assay.<b style="mso-bidi-font-weight:normal"> </b><span style="font-size:11.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:="" "times="" roman";mso-bidi-font-family:mangal;mso-ansi-language:en-gb;="" mso-fareast-language:en-us;mso-bidi-language:hi"="" lang="EN-GB">Modulatory effect of Hoechst 33258 was examined at 3 and 5 Gy using different concentrations (10, 20 and 30 <span style="mso-bidi-font-style:italic">µ<i>M</i>). Hoechst 33258 was found to inhibit radiation induced peroxidative damage and fluidity and lowered the level of <span style="font-size:11.0pt;font-family:" times="" new="" roman";="" mso-fareast-font-family:"times="" roman";mso-bidi-font-family:mangal;="" mso-ansi-language:en-sg;mso-fareast-language:en-sg;mso-bidi-language:hi"="" lang="EN-SG">nitric oxide<span style="font-size:11.0pt;font-family:" times="" new="" roman";="" mso-fareast-font-family:"times="" roman";mso-bidi-font-family:mangal;="" mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:hi"="" lang="EN-GB"> and apoptosis - as evident by DNA ladder assay and FACS, indicating free radicals scavenging potential. <span style="mso-bidi-font-style:italic">Dot plot diagramme clearly showed that 30 µ<i>M</i> 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<span style="font-size:11.0pt; font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-bidi-font-family:mangal;mso-ansi-language:en-sg;mso-fareast-language:en-sg;="" mso-bidi-language:hi"="" lang="EN-GB"> <span style="font-size:11.0pt; font-family:" times="" new="" roman";mso-fareast-font-family:"times="" roman";="" mso-bidi-font-family:mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;="" mso-bidi-language:hi"="" lang="EN-GB">and activator protein–1 was also inhibited but the antioxidant potential of the cells was enhanced.<span style="font-size:11.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:="" "times="" roman";mso-bidi-font-family:mangal;mso-ansi-language:en-sg;="" mso-fareast-language:en-sg;mso-bidi-language:hi"="" lang="EN-SG"> These findings<span style="font-size:11.0pt;font-family:" times="" new="" roman";mso-fareast-font-family:="" "times="" roman";mso-bidi-font-family:mangal;mso-ansi-language:en-gb;="" mso-fareast-language:en-us;mso-bidi-language:hi"="" lang="EN-GB"> 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.</span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span></span>
Description: 517-530
URI: http://hdl.handle.net/123456789/14482
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.50(08) [August 2012]

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