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Immunohistochemical Analysis of citrulline-nitric Oxide Cycle Enzymes and Glutamine Synthetase in Different Regions of Brain in Epilepsy Rat Model

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Neuroscience & Medicine 2014年第3期5卷 131-138页

作者： Mummedy Swamy Intan Nurfirdaus Mat Zin K. N. S. Sirajudeen Zulkarnain Mustapha Chandran Govindasamy Department of Chemical Pathology School of Medical Sciences Health Campus Universiti Sains MalaysiaKelantan Malaysia

The aim of this study was to determine the immunoreactivity of neuronal and inducible nitric oxide synthetase, argininosuccinate synthetase, argininosuccinate lyase, glutamine synthetase in different regions of brain in rats of kainic acid mediated epilepsy. Male Sprague-Dawley rats were used in this study. The acute group animals were sacrificed after 2 hours and the chronic group animals were sacrificed after 5 days of a single subcutaneous injection of kainic acid (15 mg/kg body weight). The cerebral cortex, cerebellum and brain stem slices were fixed and immunohistostained for the above enzymes. Images were captured and analyzed. In acute group, argininosuccinate synthetase and inducible nitric oxide synthetase were increased in cerebral cortex and cerebellum, neuronal nitric oxide synthetase increased in cerebral cortex and brain stem, and there was no change in argininosuccinate lyase immunoreactivity compared to control group. In chronic group, glutamine synthetase was decreased and all other enzymes immunoreactivity was increased in all the brain regions tested. This study demonstrated the up-regulation of citrul-line-nitric oxide cycle enzymes and may contribute to enhancing recycling of citrulline to arginine to support the increased production of nitric oxide in epilepsy. The decreased glutamine synthetase may increase glutamate in chronic epilepsy and may lead to neurodegeneration.

关键词： citrulline-nitric Oxide Cycle Enzymes Epilepsy Glutamine Synthetase Immunohistochemistry Kainic Acid Rat Brain

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Effects of laminar shear stress versus resveratrol on the citrulline-NO cycle in endothelial cells

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Advances in Biological Chemistry 2013年第1期3卷 18-25页

作者： Sue Im Jang Yong Chool Boo Department of Molecular Medicine Cell and Matrix Research Institute BK21 Medical Education Program for Human Resources School of Medicine Kyungpook National University Daegu Korea

Laminar shear stress (LSS) due to pulsatile blood flow enhances endothelial function by multiple mechanisms including NO production. Red wine and its constituent, resveratrol, have also been postulated to provide vascular protective effects. The aim of the present study was to compare the effects of mechanical LSS and pharmacological resveratrol treatments on the endothelial citrulline-NO cycle. Human umbilical vein endothelial cells (HUVECs) were treated with LSS (12 dyn·cm-2) or resveratrol (25 - 100 μM). The expressions of argininosuccinate synthetase 1 (ASS1), argininosuccinate lyase (ASL), nitric oxide synthase 3 (NOS3) and cationic amino acid transporter 1 (CAT1), and the production of NO were determined. The expressions of Kruppel-like factor (KLF) 2 and KLF4 as upstream regulators of ASS1 and NOS3 were also analyzed. LSS strongly increased the mRNA levels of ASS1 (8.3 fold) and NOS3 (5.4 fold) without significant effects on ASL and CAT1 mRNAs. Resveratrol increased the ASS1 mRNA level in a dose-dependent manner up to 3.8 fold at 100 μM. The effects of resveratrol on the expressions of KLF2 and KLF4 mRNAs were smaller than those of LSS. Protein levels of ASS1 and NOS3, and NO production were markedly increased by LSS but resveratrol (50 μM) increased only ASS1 protein level. The results of the current study showed that LSS had greater effects on the citrulline-NO cycle activity leading to NO production, compared to resveratrol. Because resveratrol was not so effective at stimulating the endothelial citrulline-NO cycle, further studies are needed to find more potent drugs that increase the expression of ASS1 and NOS3 genes.

关键词： citrulline-nitric Oxide Cycle Endothelial Cells Laminar Shear Stress Resveratrol

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