缝隙连接蛋白connexin26及connexin30在拟老化大鼠内耳中的表达及调控
[Abstract]:[objective] to investigate the relationship between hypermethylation in promoter region of connexin26 gene and senile deafness. [methods] D-galactose induced aging rat model was established. Auditory brainstem response (ABR) was detected by auditory brainstem response (ABR) and mitochondrial DNA4834bp deletion mutation rate was detected by real-time quantitative PCR. Real-time quantitative RT-PCR, immunoblotting Western Blot was used to detect the expression of mRNA and protein in Cx26, the promoter region of GJB2 gene (encoding Cx26 protein) was analyzed by software, the methylation degree of GJB2 gene promoter region was detected by BSP method combined with gene sequencing. Correlation analysis was used to compare the degree of methylation in the promoter region of GJB2 gene with the expression level of Cx26mRNA, protein as well as the different dose groups. [results] the mutation rate of mitochondrial DNA4834bp deletion increased with the increase of D-galactose dose, and there was no significant difference in ABR threshold shift among the four groups (P0.05). There was no significant difference in the expression of Cx26mRNA among the four groups (P0.05). The expression level of Cx26 protein in group D was significantly lower than that in group A and group B (P0.05), but there was no significant difference among the other groups (P0.05); Methyl Primer Express software showed that the promoter region of GJB2 gene was rich in CpG island). BSP-binding gene sequencing showed that the promoter region of GJB2 gene was hypermethylated in the inner ear of the high dose group, and the methylation degree of GJB2 gene was correlated with the expression of Cx26mRNA, protein. [conclusion] the hypermethylation of the promoter region of GJB2 gene is associated with its low expression in the inner ear of aging rats. It is suggested that aging methylation of GJB2 gene may be involved in the occurrence and development of senile deafness. [objective] to study the expression of gap junction protein (connexin30) in the inner ear of D-galactose-induced aging rats. [methods] D-galactose induced aging rat model was established. Auditory brainstem response (ABR) was detected by auditory brainstem response (ABR) and mitochondrial DNA4834bp deletion mutation rate was detected by real-time quantitative PCR. Real-time quantitative RT-PCR was used to detect the expression level of Cx30 mRNA and Western blot Western Blot was used to detect the expression of Cx30 protein in each group. [results] the mutation rate of mitochondrial DNA4834bp deletion increased with the increase of D-galactose dose, and there was no significant difference in ABR threshold shift among the four groups (P0.05), but there was no significant difference in Cx30mRNA expression level among the four groups (P0.05). The expression level of Cx30 protein in group B was significantly higher than that in group A and group D (P0.05), but there was no significant difference among the other groups (P0.05). [conclusion] the expression level of Cx30 protein in the inner ear tissue of aging rats increased in varying degrees, suggesting that connexin30 may be involved in the occurrence and development of senile deafness.
【学位授予单位】:华中科技大学
【学位级别】:硕士
【学位授予年份】:2011
【分类号】:R764.43
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