论文部分内容阅读
目的 通过比较年轻与年老的快速老化鼠 (senescence acceleratedmouse ,SAM)耳蜗中基因表达水平 ,筛选并分析可能的与老年聋相关的基因。方法 将分别从 2个月和 12个月的SAM耳蜗中提取的总RNA合成cDNA ,并与载有 110 1鼠基因的基因芯片进行杂交 ,杂交信号通过蓝色素 3荧光显色 ,并使用微机对其图像进行分析。通过实时定量逆转录聚合酶链式反应技术对其结果进行验证。用免疫荧光染色的方法 ,确定候选基因所编码的蛋白在耳蜗中的表达部位。结果 多数基因在老年耳蜗中的表达并没有明显的变化 ,本研究发现有 3个基因的表达较年轻耳蜗中有明显的增加 ,通过定量逆转录聚合酶链式反应技术证实其中的胸腺素beta4mRNA过度表达。免疫荧光染色显示胸腺素beta4主要在耳蜗的盖膜及外毛细胞的支持细胞中表达。结论 通过生物芯片技术 ,显示老年鼠耳蜗中基因表达的情况 ,并发现胸腺素beta4有可能与老年聋相关。这一研究为探讨老年聋的分子生物学机制提供了参考。
Objective To screen and analyze possible genes related to senile deafness by comparing the gene expression levels in the cochlea of young and old senescence accelerated mice (SAM). Methods The total RNA extracted from the cochleas of 2 months and 12 months respectively was used to synthesize cDNA. The cDNA was hybridized with the gene chip carrying 1101 murine gene. The hybridization signal was visualized by blue dye 3, Its image is analyzed. The results were validated by real-time quantitative reverse transcriptase-polymerase chain reaction. Immunofluorescence staining was used to determine the expression site of the protein encoded by the candidate gene in the cochlea. Results Most of the genes showed no significant changes in the aged cochlea. In this study, three genes were found to be significantly increased in the cochlea compared with those in the young cochlea. The excess of thymosin beta4 mRNA was confirmed by quantitative reverse transcription-polymerase chain reaction expression. Immunofluorescence staining showed that thymosin beta4 is mainly expressed in the cochlear capillaries and outer hair cells supporting cells. Conclusions Biochips showed the gene expression in the cochlea of aged mice and found that thymosin beta4 might be associated with senile deafness. This study provides a reference for exploring the molecular biological mechanism of senile deafness.