CAJ | 학술논문

目的观察氯化钴(CoCl2)对耳蜗毛细胞和神经节细胞的损伤模式并探讨Sirtuin 1(SIRT 1)在耳蜗缺氧过程中的作用.方法采用CoCl2作用于离体培养的新生大鼠耳蜗组织,通过细胞化学染色和免疫荧光染色确定CoCl2对耳蜗毛细胞存活的影响,采用实时定量PCR方法检测CoCl2作用耳蜗组织后Sirtuin 1 mRNA表达的变化.同时使用Sirtuin 1激活剂白藜芦醇和Sirtuin 1抑制剂Sirtinol,观察SIRT 1在缺氧介导的耳蜗损伤过程中的作用.结果300μmol/L以上浓度CoCl2作用于耳蜗24 h可引起内外毛细胞缺失,细胞肿胀明显,神经元胞体变得不规则,数目明显减少,有些神经纤维甚至断裂.CoCl2处理后可引起早期耳蜗组织Sirtuin 1基因表达增强,6 h达到峰值.白藜芦醇预处理可以显著提高外毛细胞(P<0.01)和内毛细胞的存活率(P<0.05).同时给予Sirtuin 1抑制剂Sirtinol,白藜芦醇的保护效应被抵消.结论 CoCl2导致耳蜗组织缺氧损害,耳蜗在缺氧过程中Sirtuin 1早期上调,可保护耳蜗组织对抗缺氧诱导的细胞损伤.
목적관찰록화고(CoCl2)대이와모세포화신경절세포적손상모식병탐토Sirtuin 1(SIRT 1)재이와결양과정중적작용.방법채용CoCl2작용우리체배양적신생대서이와조직,통과세포화학염색화면역형광염색학정CoCl2대이와모세포존활적영향,채용실시정량PCR방법검측CoCl2작용이와조직후Sirtuin 1 mRNA표체적변화.동시사용Sirtuin 1격활제백려호순화Sirtuin 1억제제Sirtinol,관찰SIRT 1재결양개도적이와손상과정중적작용.결과300μmol/L이상농도CoCl2작용우이와24 h가인기내외모세포결실,세포종창명현,신경원포체변득불규칙,수목명현감소,유사신경섬유심지단렬.CoCl2처리후가인기조기이와조직Sirtuin 1기인표체증강,6 h체도봉치.백려호순예처리가이현저제고외모세포(P<0.01)화내모세포적존활솔(P<0.05).동시급여Sirtuin 1억제제Sirtinol,백려호순적보호효응피저소.결론 CoCl2도치이와조직결양손해,이와재결양과정중Sirtuin 1조기상조,가보호이와조직대항결양유도적세포손상.
Objective To observe the pattern of damage to cochlear hair cells and spiral ganglion neurons(SGN) induced by CoCl2 and to explore the role of the Sirtuin 1 (SIRT 1)in the process of cochlear hypoxia. Methods The rat cochlear tissues were isolated and cultured, and treated with different concentrations of CoCl2. Cytochemical and immunofluorescence staining were performed to determine the effects of CoCl2 on hair cells and SGNs. Sirtuin 1 gene change was detected by real time PCR assay at different time points after CoCl2 treatment. Resveratrol, an activator of Sirtuin 1, and Sirtinol, an inhibitor of Sirtuin 1, were used to observe the role of Sirtuin 1 in the process of hypoxia-mediated cochlear damage. Results 300 μmol / L or higher concentrations of CoCl2 can cause inner and outer hair cell loss in the cochlear explants, cell swelling, irregular shapes of SGN bodies, the decreasing number of SGNs or even nerve fiber rupture. Sirtuin 1 gene expression in cochlear explants was up-regulated at early stage after CoCl2 treatment. Resveratrol pretreatment can significantly improve the viability of outer hair cells (P<0.01) and inner hair cells (P<0.05). The protective effects of resveratrol can be offset by Sirtinol. Conclusion CoCl2 can cause hypoxic damage in cochlear explants and up-regulate Sirtuin 1 gene expression at the early stage. Sirtuin 1 may play a protective role against hypoxia-induced cochlear cell damage.