叔丁基双氧水诱导HEI-OC1细胞株凋亡与STATs mRNA表达的相关性分析

Analysis of the Relevance between t-BHP Induced Apoptosis and STATs mRNA Expression in HEI-OC1 Cells

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。

Objective To study the biological effects of(Janus Kinase-Signal transducers and activators of transcription,AK-STATs)path-way in hair cell apoptosis by analyzing the expression level of signal transducers and activators of transcription(STATS)of apoptotic HEI-OC1 cells line induced by oxidative stress injury.Methods Using different poisoning concentrations(0,20,40,60μM)of(Tert-Butyl hydroperoxide,T-BHP)built the apoptotic HEI-OC1 cells model while 0μM was used as the control group.The apoptosis level at different poisoning concentration groups was detected by Annexin V-FITC/PI and the expression level of STATs mRNA by real-time quantitative PCR detecting system.Results The apoptosis rates in different contamination groups(0,20,40,60μM)were 3.9%,7.4%,32.0%,and 91.2%,respectively.Compared with control group,STAT1 mRNA,STAT3 mRNA,STAT5 mRNA,and STAT6 mRNA expression levels in different poisoning concentration groups declined significantly(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).No statistically significant differences were found among the poisoning concentration groups(P>0.05).Compared with the control group,the STAT2 mRNA expression levels changed significantly(F=1 259.148,P<0.01).The STAT2 mRNA expression level in 20μM contamination group decreased while 40,60μM contamination group increased significantly(P<0.01).No expression of STAT4 in HEI-OC1 cell was noted.Conclusion The JAK-STAT2 signal pathway has the biological effects on leading oxidative stress injury apoptosis and JAK-STAT1/STAT3/STAT5/STAT6 has the biological effect of inhibit oxidative stress injury apoptosis.