ISSN: 2161-0460

Journal of Alzheimers Disease & Parkinsonism
Open Access

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Research Article

Generation of Reactive Oxygen Species (Ros) and Pro-Inflammatory Signaling in Human Brain Cells in Primary Culture

Walter J. Lukiw1, Surjyadipta Bjattacharjee1, Yuhai Zhao2, Aileen I. Pogue3 and Maire E. Percy4*

1LSU Neuroscience Center and Department of Ophthalmology, Louisiana State University Health Sciences Center, New Orleans LA 70112, USA

2University of Texas Health Science Center, Houston TX 77030, USA

3Alchem Biotek Corporation, Toronto ON M5T 1L8, Canada

4Surrey Place Centre, Toronto ON M5S 2C2 and University of Toronto Departments of Physiology and Obstetrics & Gynaecology, Toronto ON M5S 1A8, Canada

Corresponding Author:
Maire E. Percy
Neurological Disease Research Group
LSU Neuroscience Center and Department of Ophthalmology
Louisiana State University Health Sciences Center 2020 Gravier Street
New Orleans LA 70112 USA
Email: wlukiw@lsuhsc.edu

Received date: November 26, 2011; Accepted date: January 23, 2012; Published date: January 25, 2012

Citation: Lukiw WJ, Bjattacharjee S, Zhao Y, Pogue AI, Percy ME (2012) Generation of Reactive Oxygen Species (ROS) and Pro-Inflammatory Signaling in Human Brain Cells in Primary Culture. J Alzheimers Dis S2:001. doi:

Copyright: © 2012 Lukiw WJ, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

The cellular generation of reactive oxygen species (ROS) has been implicated in contributing to the pathology of human neurological disorders including Alzheimer’s disease (AD) and Parkinson’s disease (PD). To further understand the triggering and participation of ROS-generating species to pro-inflammatory and pathological signaling in human brain cells, in these experiments we studied the effects of 22 different substances (including various common drugs, interleukins, amyloid precursor protein, amyloid peptides and trace metals) at nanomolar concentrations, in a highly sensitive human neuronal-glial (HNG) cell primary co-culture assay. The evolution of ROS was assayed using the cell-permeate fluorescent indicator 2’,7’-dichlorofluorescein diacetate (H2DCFDA), that reacts with major ROS species, including singlet oxygen, hydroxyl radicals or superoxides (λEx 488 nm; λEm 530 nm). Western analysis was performed for cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2) and cytosolic phospholipase A (cPLA2) to study the effects of induced ROS on inflammatory gene expression within the same brain cell sample. The data indicate that apart from acetylsalicylic acid (aspirin) and simvastatin, several neurophysiologically-relevant concentrations of Aβpeptides and neurotoxic trace metals variably induced ROS induction, COX-2 and cPLA2 expression. These findings have mechanistic implications for ROS-triggered inflammatory gene expression programs that may contribute to AD and PD neuropathologic mechanisms.

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