三磷酸腺苷在新生大鼠耳蜗血管纹缘细胞中的释放机制  

Mechanisms of ATP Release from Cochlear Stria Vascularis Marginal Cells in Neonatal Rats

作　　者：刘斌[1] 李吉平[1] 刘君[1] LIU Bin;LI Jiping;LIU Jun(Department of Otorhinolaryngology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China)

机构地区：[1]上海交通大学医学院附属仁济医院耳鼻咽喉科

出　　处：《中华耳科学杂志》2019年第3期394-399,共6页Chinese Journal of Otology

基　　金：国家自然科学基金青年科学基金项目(No.81600797);上海交通大学医工交叉面上项目(YG2017MS49)~~

摘　　要：目的探究新生大鼠耳蜗血管纹缘细胞中ATP释放的机制。方法原代培养新生SD大鼠血管纹缘细胞,利用钙离子荧光探针、生物发光法测定ATP浓度和β-氨基己糖苷酶释放率测定法来研究ATP和GPN与P2YR-PLC-IP3通路诱导的内质网钙库反应之间的关系,以及内质网钙库与细胞外ATP浓度和β-氨基己糖苷酶释放率的关系。结果 ATP和GPN可通过P2YR-PLC-IP3通路诱发内质网钙库释放,可被P2YR-PLC-IP3通路拮抗剂抑制。缘细胞外ATP浓度和β-氨基己糖苷酶释放率与细胞内钙离子浓度呈正相关。结论缘细胞外ATP作用于P2Y嘌呤能信号系统触发内质网钙库释放,胞内钙离子诱导溶酶体胞吐作用释放ATP至胞外,从而发挥其生物学作用。Objective To elucidate mechanisms of ATP release from cochlear stria vascularis marginal cells in neonatal rats. Methods Stria vascularis marginal cells from 1 day old Sprague Dawley rats were cultured. Fluo-4 AM was used to investigate the influence of ATP and GPN on P2YR-PLC-IP3 pathway induced endoplasmic reticulum(ER) Ca2+ responses. ATP bioluminescence assay and β-hexosaminidase secretion assay were used to investigate the relationship between ER Ca2+ responses and extracellular ATP and β-hexosaminidase. Results ATP and GPN induced Ca2+ release from ER via the P2YR-PLC-IP3 pathway, which was inhibited by P2Y receptor antagonists. Release of ATP and β-hexosaminidase from marginal cells were dependent on ER Ca2+ responses. Conclusion Extracellular ATP can evoke ER Ca2+ responses via the P2Y-PLC-IP3 pathway. ER Ca2+ contributes to ATP release from lysosomes via exocytosis which helps to control Ca2+ homeostasis in marginal cells.

关 键 词：三磷酸腺苷 溶酶体 P2YR-PLC-IP3通路 

分 类 号：R764[医药卫生—耳鼻咽喉科]