Date of Award

2013

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemical Engineering

First Advisor

Zhu, Donghui Dr.

Abstract

The occurrence of neurodegenerative diseases (NDs) increases with extended life expectancy. The rise in elderly populations is resulting in an increased incidence of cognitive impairment, and a high risk for developing dementia. Oxidative stress is implicated in the pathogenesis of a number of neurovascular disorders, such as Alzheimer’s disease (AD) and Parkinson’s disease (PD). It has also been linked to disruption of blood brain barrier (BBB) in hypoxic-ischemic injury. As part of the neurovascular unit, pericytes are inevitably subjected to insult from oxygen and nitrogen radicals, generated in the central nervous system (CNS), leading to cell dysfunction or damage. In this research, we studied the effect of oxidative stress on cerebro-vascular pericytes cellular function and determined how oxidative stress changes pericytes protein expression, including F-actin, α-actin, platelet-derived growth factor receptor beta (PDGFR-β), tropomyosin and myosin. In vitro, human brain vascular pericytes cell cultures were exposed to hydrogen peroxide (H2O2) to mimic the cerebral ischemic condition then evaluate the cytotoxicity by MTT viability assay, LDH assay and quantity of cellular morphology deterioration. Intracellular Proteins expression level was examined by quantitative immunefluorescent microscopy. Proteins were labeled with specific primary antibodies and corresponding secondary antibodies, conjugated with different fluorescent probes. Results from cytotoxicity tests suggest that when we exposed the pericytes cells to 100 and 200 μM for 1 hour, it showed the toxic effect of H2O2 on the pericytes cells without killing the cells. Our data reveals that after mild oxidative stress, pericytes protein expression intensity increased, changed position and moved toward the cell membrane while sometimes forming cellular protrusion connecting cells together. These results indicate that oxidative stress can affect pericytes cell contraction and cell movement.

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