Acute Vibration Induces Oxidative Stress And Changes In Transcription In Soft Tissue Of Rat Tails - Introduction; Proceedings Of The First American Conference On Human Vibration
Description:Repeated exposure to hand-arm vibration through the use of vibrating hand tools can result in the development of the disorder known as hand-arm vibration syndrome (HAVS; (1,3)). One of the hallmark symptoms of HAVS is cold-induced peripheral vasospasms that result in finger blanching (4). Although the vascular and neural pathology associated with vasospasms has been described, little is known about cellular mechanisms leading to this damage (4). To understand how vibration may alter vascular and neural physiology and anatomy, rats were exposed to a single bout of tail vibration and the molecular responses of neural and vascular tissues were measured to determine if there are immediate or sustained affects of vibration that may underlie longer term changes in physiology. Methods Experiment 1. Male Sprague Dawley rats (n = 32, 6 weeks of age) were housed in AAALAC accredited facilities. All procedures were approved by the NIOSH Animal Care and Use Committee and were in compliance with the CDC guidelines for care and use of laboratory animals. Vibration exposures were performed by restraining rats in Broome-style restrainers, and securing their tails to a vibration platform using 6 mm wide straps that were placed over the tail every 3 cm. Restraint control rats were treated in the same manner, except that the tail platform was mounted on isolation blocks and not on a shaker. The vibration exposure was 125 Hz, 49 m/s2, for 4 h. Rats were euthanized with an overdose of pentobarbital (100 mg/kg) 1 h or 24 hours after the exposure. RTqPCR was used to measure transcript levels for endothelin 1 (ET-1), the 3 forms of nitric oxide synthetase (NOS) NOS-1, NOS-2, NOS-3 and norepinephrine receptor subtypes 1D, 2A, 2C, in artery tissue, and to measure calcitonin gene-related peptide (CGRP) and nitric oxide synthase-1 (NOS-1) in ventral nerves. Data were analyzed using 2-way ANOVAs. Experiment 2. Male Sprague Dawley rats (n =24, 6 weeks of age) were maintained as described above. Animals were exposed to a single bout of restraint or vibration. Another group of animals served as cage controls. All animals were euthanized 24 h after the exposure, and tail arteries were isolated and frozen. Reactive oxygen species ROS) were measured using electron spin resonance spectroscopy (ESR). Arteries were homogenized over ice in 1 ml of PBS with protease inhibitor cocktail, using a tissue tearer (Biospecs Products Inc. Racine, WI USA). The sample was split into two 0.5 ml samples. One set had PBS and the spin label hydroxyl-TEMPO [0.1 mM] added while the other set had hydroxyl-TEMPO [1.0 mM] plus the specific hydroxyl radical scavenger, dimethylthiourea (DMTU), added to confirm the presence of the hydroxyl radical. Samples were vortexed and then placed in a flat cell for ESR analysis. The ESR spectrometer settings were: receiver gain, 6.32 x 102; time constant, 0.02 s; modulation amplitude, 1.0 G; scan time, 20 sec; magnetic field, 3490 ± 100 G (2). Data were analyzed using 1-way ANOVAs.
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