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A Study on Expression of Heat Shock Protein 70 in Rats Cochleae with Sodium Salicylate

Author: YangNing
Tutor: JiangXueJun
School: China Medical University
Course: Otorhinolaryngology
Keywords: Sodium salicylate Heat shock protein Cochlea Immunohistochemistry Ototoxicity
CLC: R764
Type: Master's thesis
Year: 2002
Downloads: 29
Quote: 0
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Abstract


Preface heat shock protein is a highly conserved protein, by a variety of physical, chemical, and biological stressors induce the synthesis, plays an important role in the conservation organization, cell tolerance ischemia, hypoxia or toxic injury. Oxidative damage, the height of the heat shock protein expression can protect cells against oxidative damage enhanced ischemic tolerated. As an endogenous protective mechanism, heat shock protein achieve a more natural protection than drugs. In otologic research showed that the cochlea of ??heat shock protein expression increased significantly reduces the strong noise stimulation induced permanent threshold shift. Induction of heat shock protein expression and thus how effective the recent hot research. Especially in conditions without causing cell damage induced more application prospects or potential induced synthesis of HSP. Salicylic acid drugs its good efficacy of countries in the world are widely used. However, large doses of salicylic acid drugs will produce a transient hearing loss and tinnitus, considered an ototoxic drugs. Recently found that sodium salicylate can reduce hearing loss caused by noise and gentamicin. Another experiment found that sodium salicylate can promote the expression of heat shock proteins. Taking into account the protective effect of heat shock proteins, we speculate that sodium salicylate may also induce heat shock proteins in the cochlea. Therefore, this experiment as the starting point of sodium salicylate in the cochlea induction of heat shock proteins, and to explore the mechanism of sodium salicylate on cochlear provide experimental evidence and theoretical basis for its application in otology. MATERIALS AND METHODS Materials: Take the auricle reflex normal healthy male Wistar rats 57, weighing 220-260 g, were randomized. 100mg/kgSS group: 100 mg / kgSS, ip; 250mg/kgSS group: 250 mg / kgSS, ip; 500mg/kgSS group: 500 mg / kgSS, ip; control group: normal saline lP; 37 ° C group: a 37 ° C environment, and after 30 minutes at room temperature under restoration; 100 mg / kgSS +37 ° C group: 100 mg / kgSS, LP, 60 minutes descendants 37 ° C environment for 30 minutes, then at room temperature restored. These groups of nine. The positive control group (n = 3), heat shock disposal. Methods: door) ABR detection: In addition to the positive control group, each group 3 before treatment and after 4 hours and 24 hours the rats with 3% sodium pentobarbital 40mg/kg intraperitoneal anesthesia test ABR threshold. The issued the frequency the sound based CliCk to ZKHZ stimulus signal Danac-7 type audiostimulator, test strength by 95dB SPL, close to the threshold of 5 dB by-file diminishing, the observation dish waves to determine the threshold of hearing. Flash) immunohistochemical techniques: the remaining animals drawn six hours after treatment. Removed after perfusion fixation bulla on the same 4 ° C in 4% paraformaldehyde, anatomical Abstract aluminum bone under a microscope to pierce the round window cochlear top pick open at the same fixative 4 ° C for 24 hours, within perfusion. With PBS decalcified in formic acid - sodium formate decalcification solution at room temperature after 7 days, the daily replacement decalcification liquid. And then rinsed with PBS, 30% sucrose 4T overnight settlement underwent OCT embedding constant cold box slicer along modiolus consecutive frozen sections, slice thickness of 5 pm. By immunohistochemistry the (h footwork) HSwo staining. First slices were incubated with 0.5% hydrogen peroxide at room temperature, dropping A solution (mouse anti-human monoclonal antibody) and B solution (goat anti-mouse IgG biological complexing agent DAB color gradient alcohol dehydration Fengpian. To microscopic images mean optical density value of all experimental data analysis system to calculate the heat shock protein-positive reaction product to the standard deviation of the mean soil (hi s) said the use of statistical analysis the SPSS10.0 package handling, Dunnett t test and SNK test experimental results a 1.250mg/kgSS group, and 55mg/kgSS-group after the injection of sodium salicylate in rats hearing a temporary impact, four hours after treatment (P o decreased hearing. 05 people in other groups before treatment, after ** R threshold was no significant difference in sexual u knock .05 2 normal rat cochlear outer hair cells were positive, stria vascularis, spiral ligament expression · 2 · previously spiral ligament court order spiral ligament and tympani spiral ligament coloring heavier 100mgikgSS group compared with the control group, the difference was not statistically significant. 250mg/kgSS group of outer hair cells and the expression of the vascular pattern at significantly increased, and the difference was the significance. 37t group at the stria vascularis expression increased statistical significance. k 500mg/kgSS group and 100m mine furnace S +37 ℃ various parts of the group differences were statistically significant. discussion of the heat shock protein is a class of widespread, evolutionarily highly conserved protein. organisms by non-lethal biological, chemical or physical factors such as stimulus to produce heat shock response, mainly for the induction of heat shock protein and normal protein synthesis inhibition and the heat shock response with cross-tolerance phenomenon from its foundation is the up-regulation of heat shock protein expression. many family of heat shock proteins, especially HSP70 family is one of the important active ingredients, the exact role of the protection and extensive clinical significance. structured HSpoo usually unstressed cells there in expression, as observed in this experiment, the normal stress of outer hair cells showed a positive reaction, the lower expression of vascular stria and spiral ligament, spiral ligament, spiral ligament court order and drums order spiral ligament blush slightly heavier normal rat cochlea the lower structure HSpoo expression of gene regulation in body under stress injury, a large number of synthetic stress-induced type of HSP70 in a variety of injuries play an important role in the induction of tolerance The experiment found that sodium salicylate 500mg/kg group, the outer hair cells of the cochlea, stria vascularis and spiral ligament at the immune response strength is significantly enhanced. sodium salicylate 250mg/kg The group also observed that the immune response of the outer hair cells and vascular pattern at significantly enhanced both groups lead to hearing a transient decline in cell toxic dose of sodium salicylate directly induced HSP70 synthesis. sodium salicylate injury the main site of the outer hair cells and vascular pattern, to coincide with this change of the strongest parts of the experiment in the immune response. sodium salicylate 100mg/kg group was not observed to have significant differences in the immune response and the control group, ABR threshold also change significantly non fine?

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CLC: > Medicine, health > Otorhinolaryngology > Otology,ear disease
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