目的通过分析氧化应激损伤的凋亡HEI-OC1细胞各信号转导及转录激活子(signal transducers and activators of transcription,STATs)的表达,探讨酪氨酸激酶-信号转导及转录激活子(janus kinase-signal transducers and activators of tra...目的通过分析氧化应激损伤的凋亡HEI-OC1细胞各信号转导及转录激活子(signal transducers and activators of transcription,STATs)的表达,探讨酪氨酸激酶-信号转导及转录激活子(janus kinase-signal transducers and activators of transcription,JAK-STATs)信号通路在毛细胞凋亡中的生物学作用。方法采用不同浓度(0、20、40、60 μM)叔丁基过氧化氢(tert-butyl hydroperoxide,t-BHP)染毒构建HEI-OC1细胞株凋亡模型(0μM浓度为对照组),Annexin V-FITC/PI法检测20、40、60 μM浓度t-BHP染毒组细胞凋亡水平,实时荧光定量PCR检测STATs mRNA表达水平。结果对照组及20、40、60 μM t-BHP染毒组的细胞凋亡率分别为3.9%、7.4%、32.0%、91.2%;与对照组相比,各浓度T-BHP染毒组STAT1mRNA、STAT3mRNA、STAT5mRNA、STAT6 mRNA的表达水平均显著性下降(分别为F=5 534.302,P<0.01;F=146.038,P<0.01;F=685.929,P<0.01;F=516.11,P<0.01),各浓度t-BHP染毒组间差异无统计学意义(P>0.05);与对照组相比各浓度t-BHP染毒组STAT2mRNA的表达水平均有显著变化(F=1 259.148,P<0.01),20 μM染毒组STAT2 mRNA的表达水平明显下降(P<0.01),但40、60 μM染毒组均明显升高(P<0.01);STAT4 mRNA在HEI-OC1细胞株中未见表达。结论 JAK-STAT2信号通路具有促进毛细胞氧化应急损伤凋亡的生物学作用,JAK-STAT1/STAT3/STAT5/STAT6信号通路有抑制毛细胞氧化应急损伤凋亡的生物学作用。展开更多
Objective Age-related hearing loss(AHL),characterized by degeneration of cochlea structures,is the most common sensory disorder among the elderly worldwide.The calcium channel is considered to contribute to normal hea...Objective Age-related hearing loss(AHL),characterized by degeneration of cochlea structures,is the most common sensory disorder among the elderly worldwide.The calcium channel is considered to contribute to normal hearing.However,the role of the T-type voltage-activated calcium channel,Cav3.1,remains unclear in AHL.Here,we investigate the age-related change of Cav3.1 expression in the cochlea and D-gal-induced senescent HEI-OC1 cells.Methods Cochleae from C57BL/6 mice at 2 months and 12 months of age were assessed.Senescence in House Ear Institute-Organ of Corti 1(HEI-OC1)cells was induced by D-gal treatment.The immunofluorescence technique was employed to investigate the distribution of Cav3.1 in vivo and in vitro.Quantitative assessment was achieved by Western blotting and real-time PCR.Results In comparison with 2-month-old animals,12-month old C57BL/6 mice exhibited great loss of hair cells and elevated auditory brainstem threshold.The Cav3.1 was located in hair cells,spiral ganglion cells,lateral walls,and the expression of Cav3.1 protein and mRNA decreased in the aged cochleae.D-gal-induced senescence assay confirmed the down-regulation of Cav3.1 expression in senescent HEI-OC1 cells.Conclusion Our results show that age-related down-regulated expression of Cav3.1 in the cochleae is associated with AHL and may contribute to the pathogenesis of AHL.展开更多
基金supported by grants from the National Natural Sciences Foundation of China(No.81800911 and No.81771004)the Anhui Natural Science Foundation(No.1808085QH248)Fundamental Research Funds for the Central Universities(No.WK9110000053).
文摘Objective Age-related hearing loss(AHL),characterized by degeneration of cochlea structures,is the most common sensory disorder among the elderly worldwide.The calcium channel is considered to contribute to normal hearing.However,the role of the T-type voltage-activated calcium channel,Cav3.1,remains unclear in AHL.Here,we investigate the age-related change of Cav3.1 expression in the cochlea and D-gal-induced senescent HEI-OC1 cells.Methods Cochleae from C57BL/6 mice at 2 months and 12 months of age were assessed.Senescence in House Ear Institute-Organ of Corti 1(HEI-OC1)cells was induced by D-gal treatment.The immunofluorescence technique was employed to investigate the distribution of Cav3.1 in vivo and in vitro.Quantitative assessment was achieved by Western blotting and real-time PCR.Results In comparison with 2-month-old animals,12-month old C57BL/6 mice exhibited great loss of hair cells and elevated auditory brainstem threshold.The Cav3.1 was located in hair cells,spiral ganglion cells,lateral walls,and the expression of Cav3.1 protein and mRNA decreased in the aged cochleae.D-gal-induced senescence assay confirmed the down-regulation of Cav3.1 expression in senescent HEI-OC1 cells.Conclusion Our results show that age-related down-regulated expression of Cav3.1 in the cochleae is associated with AHL and may contribute to the pathogenesis of AHL.
