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Detection and Localization of HSP27、Scn8a in the Rat Cochlea after Sodium Salicylate Injection

Author: FanQiJun
Tutor: HuangZhiWu
School: Wuhan University
Course: Otorhinolaryngology
Keywords: sodium salicylate cochlea RTQ RT-PCR HSP27 Scn8a
CLC: R764.45
Type: Master's thesis
Year: 2005
Downloads: 39
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Abstract


The mechanism of tinnitus is not clearly understood. As we now known, the animal model of tinnitus induced by sodium salicylate is the basical tool of tinnitus research. W e have used gene chip technique to analyze the gene expression profile changes of cochleas in rats induced by single high dose salicylate injection and long term injection in early research. In order to know more about mechanism of tinnitus, in this research, some genes were chosen to be validated by using real time RT-PCR with SYBR GREEN I according as the results of gene expression profile and localization of production of one gene in the rat cochlea.Objective To investigate the genes expression of HSP27 and Scn8a in cochlea of rats induced by long term and single high dose sodium salicylate injection and discuss its role in sodium salicylate ototoxity.Methods The level of HSP27 and Scn8a genes expression in specimens was measured using real time quantitative reverse transcription PCR (RTQ RT-PCR) with SYBR GREEN I. β-actin was used as internal control. The expression of HSP27 in rats cochlea induced by long term sodium salicylate was examined by immunohistochemistry and image analysis system.Results The gene expression of HSP27 in cochlea of rats induced by long term sodium salicylate injection was significantly increased compared to that of the normal rats. (P<0.01) The gene expression of Scn8a in cochlea of rats induced by single high dose sodium salicylate injection was significantly decreased compared to that of the normal rats. (P<0.01) The HSP27 had been stained in the cochlear , especially in the organ of Corti, spiral ganglion , stria vascularis, and spiral ligand.The expression of HSP27 was significantly enhanced at each of the above mentioned position after sodium salicylate injection (P<0.01)and the enhance at Corti’s organ was more obvious.Conclusion RTQ RT-PCR with SYBR GREEN I fluorescence can be a reliable method for validating the results of gene expression profile. Long term sodium salicylate injection has an obvious inducing effect on the gene expression of HSP27 in rat’s cochlea . The gene expression of Scn8a in cochlea of rats induced by single high dose sodium salicylate injection was significantly decreased. HSP27 is selectively expressed in rat’s cochlea. It is revealed that HSP27 may play an important role in thedevelopment of tinnitus.As a result, we found RTQ RT-PCR with SYBR GREEN I fluorescence would be a reliable method for those rarely available tissues to detect the gene expression and localization of production of genes in the rat cochlea by immunohistochemistry . The study can provide insights into mechanism of tinnitus.

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CLC: > Medicine, health > Otorhinolaryngology > Otology,ear disease > Ear nervous system diseases > Tinnitus
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