血红素加氧酶-1在硫化氢对抗顺铂诱导的听细胞毒性损伤中的作用

摘要

目的 探讨血红素加氧酶-1(HO-1)在硫化氢(H2S)对抗顺铂(Cisplatin)诱导的听细胞损伤中的作用.方法 应用顺铂处理听细胞,建立化疗药物耳毒性的体外模型.硫化氢的供体硫氢化钠( NaHS)及HO-1抑制剂锌原卟啉Ⅸ(ZnPPⅨ)先于顺铂加入培养基中,作为预处理.检测细胞存活率、胞内丙二醛( MDA)的含量、HO-1的表达.结果 顺铂在10～80μmol/L浓度范围内处理听细胞24h可浓度依赖性地降低细胞存活率.40μmol/L顺铂处理24 h可增加听细胞内MDA的含量.NaHS处理可上调HO-1的表达,抑制顺铂引起的胞内MDA含量增加及细胞存活率降低.HO-1的抑制剂棗ZnPPⅨ可明显减弱H2S对顺铂诱导的胞内MDA含量的增加及细胞存活率降低的抑制作用.结论 HO-1可通过抗氧化的机制介导H2S对顺铂诱导的听细胞毒性的改善作用.%Objective To explore the roles of beme oxygenase 1 ( HO-1 ) in the protection of hydrogen sulfide ( H2S ) against cisplatin-induced injuries in auditory cells.Methods Auditory cells were treated with cisplatin to establish a chemotherapeutics-inducel ototoxicity model.Sodium hydrosulfide ( NaHS,a H2S donor )was added into medium for 1 h before cisplatin treatment.To inhibit the role of HO-1,zine protoporphyrin IX ( ZnPPIX )was used at the time of treatment with NaHS and cisplatin.Cell viabihty was tested by using the cell counting kit ( CCK-8 ).Level of intercellular MDA was detected by a commercial kit.The expressions of HO-1 and cleaved caspase-3 were detected by Western blot.Apoptotic rate was determined by propidium iodide staining and flow cytometry.Results Cisplatin at 10～80 μmol/L for 24h significantly induced cytotoxicity,reducing cell viability.Treatment with cisplatin of 40 μmol/L for 24 hincreased the level of intercellular MDA,expression of cleaved caspase-3,and rate of cellular apoptosis.Pretreatment with NaHS at different concentrations dramatically attenuated the decrease in cell viability induced by cisplatin,and also inhibited an increase in intercellular MDA level,expression of cleaved caspse-3,and rate of apoptosis induced by cisplatin.By inhibiting the role of HO-1,ZnPPIX blocked the inhibitory effects of H2S on an increase in intercellular M DA induced by cisplatin,overexpression of cleaved caspase-3,and apoptosis.Conclusions HO-1 may mediate auditory cells protection of H2S via inhibiting oxidative stress and cisplatin- induced apoptosis.