中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2016, Vol. 11 ›› Issue (7): 1134-1140.doi: 10.4103/1673-5374.187051

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

热休克蛋白70保护缺血缺氧PC12细胞的机制：维持细胞内Ca2+稳态

  

  • 出版日期:2016-07-30 发布日期:2016-07-30
  • 基金资助:
    中国国家自然科学基金项目（81571938，81501706）；山东省自然科学基金项目（Y2007C133）

Heat shock protein 70 protects PC12 cells against ischemia-hypoxia/reoxygenation by maintaining intracellular Ca2+ homeostasis

Yuan Liu1, Xue-chun Wang1, Dan Hu1, Shu-ran Huang2, Qing-shu Li1, Zhi Li1, Yan Qu1, *   

  1. 1 Department of Intensive Care Unit, Affiliated Qingdao Municipal Hospital of Qingdao University, Qingdao, Shandong Province, China 2 Department of Intensive Care Unit, Affiliated Hospital of Jining Medical University, Jining, Shandong Province, China
  • Online:2016-07-30 Published:2016-07-30
  • Contact: Yan Qu, qdquyan@aliyun.com.
  • Supported by:
    This research was supported by the National Natural Science Foundation of China, No. 81571938 and 81501706; the Natural Science Foundation of Shandong Province of China, No. Y2007C133.

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摘要：

热休克蛋白70可以高度保持PC12细胞内钙稳态，可能是抗凋亡的重要机制之一，但其具体的影响机制尚不明确。为此实验设计在缺氧复氧损伤PC12细胞中以慢病毒载体转染外源性热休克蛋白70基因，观察Ca2+水平的变化。结果发现，当热休克蛋白70过度表达时，缺氧复氧的神经元存活率和ATP酶活性增加，细胞活性氧水平和细胞内Ca2+浓度降低，且内质网（肌浆网）Ca2+-ATP酶蛋白和mRNA表达增强，1，4，5-三磷酸肌醇受体蛋白和mRNA表达下降，细胞内Ca2+浓度降低。说明外源性热休克蛋白70过表达能通过上调内质网（肌浆网）Ca2+-ATP酶蛋白表达同时下调1，4，5-三磷酸肌醇受体蛋白来稳定Ca2+稳态，继而发挥对缺氧复氧损伤神经元的保护作用。 

orcid: 0000-0003-1166-055X (Yan Qu)

关键词: 神经再生, 外源性热休克蛋白70, 慢病毒转染, 缺血缺氧, PC12细胞, Ca2+, 内质网, 三磷酸肌醇受体, 内质网（肌浆网）Ca2+-ATP酶

Abstract: Heat shock protein 70 (HSP70) maintains Ca2+ homeostasis in PC12 cells, which may protect against apoptosis; however, the mechanisms of neuroprotection are unclear. Therefore, in this study, we examined Ca2+ levels in PC12 cells transfected with an exogenous lentiviral HSP70 gene expression construct, and we subsequently subjected the cells to ischemia-hypoxia/reoxygenation injury. HSP70 overexpression increased neuronal viability and ATPase activity, and it decreased cellular reactive oxygen species levels and intracellular Ca2+ concentration after hypoxia/reoxygenation. HSP70 overexpression enhanced the protein and mRNA expression levels of sarcoplasmic/ endoplasmic reticulum Ca2+-ATPase (SERCA), but it decreased the protein and mRNA levels of inositol 1,4,5-trisphosphate receptor (IP3R), thereby leading to decreased intracellular Ca2+ concentration after ischemia-hypoxia/reoxygenation. These results suggest that exogenous HSP70 protects against ischemia-hypoxia/reoxygenation injury, at least in part, by maintaining cellular Ca2+ homeostasis, by upregulating SERCA expression and by downregulating IP3R expression.

Key words: nerve regeneration, exogenous heat shock protein 70, lentivirus transfection, ischemia-hypoxia/reoxygenation, PC12 cells, Ca2+, endoplasmic reticulum, inositol 1,4,5-trisphosphate receptor, sarcoplasmic/endoplasmic reticulum Ca2+-ATPase, neural regeneration