三磷酸腺苷在新生大鼠耳蜗血管纹缘细胞中的释放机制.pdf

中华耳科学杂志2019年第17卷第3期 ·394 · ·基础研究 · 三磷酸腺苷在新生大鼠耳蜗血管纹缘细胞中的释放机制 刘斌 李吉平 刘君* 上海交通大学医学院附属仁济医院耳鼻咽喉科（上海200120） 【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。 【关键词】三磷酸腺苷；溶酶体；P2YR-PLC-IP3通路 【中图分类号】R764 【文献标识码】A 【文章编号】1672-2922（2019）03-394-6 Mechanisms of ATP Release from Cochlear Stria Vascularis Marginal Cells in Neonatal Rats LIU Bin , LI Jiping , LIU Jun* Department of Otorhinolaryngology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China Corresponding author: LIU Jun Email: liujun1220@ 【Abstract 】Objective To elucidate mechanisms of ATP release from cochlear stria vascularis marginal cells in neo- natal rats. Methods Stria vascularis marginal cells from 1 day old Sprague Dawley rats were cultured. Fluo-4AM 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 ß-hexosaminidase secretion assay were used to investigate the relationship between ER Ca2 + responses and extracellular ATP and ß-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 ß-hex- osaminidase from marginal cells w