FK506协同NGF促进缺氧神经细胞突起生长

FK506协同NGF促进缺氧神经细胞突起生长 Combined Treatment with FK506 and Nerve Growth Factor for Neurite Outgrowth after Hypoxia 张珠 ZHANG Zhu 1 first-author 张开明 ZHANG Kaiming 2 刘芳 LIU Fang 2 殷义霞 YIN Yixia 3 李世普 LI Shipu 3 武汉理工大学校医院 武汉理工大学校医院 Affiliated Hospital of Wuhan University of Technology Affiliated Hospital of Wuhan University of Technology 岳阳市二人民医院骨科岳阳市骨科重点实验室 岳阳市二人民医院骨科岳阳市骨科重点实验室 Department of Orthopedic Surgery,the Second People’s Hospital of Yueyang City Department of Orthopedic Surgery,the Second People’s Hospital of Yueyang City 武汉理工大学材料复合新技术国家重点实验室 武汉理工大学材料复合新技术国家重点实验室 State Key Laboratory of Advanced Technology for Materials Synthesis and Processing,Wuhan University of Technology State Key Laboratory of Advanced Technology for Materials Synthesis and Processing,Wuhan University of Technology 为探讨免疫抑制药物FK506对缺氧神经细胞的影响和相关保护机制,构建了神经细胞缺氧模型,对比分析并定量检测了常氧与缺氧状态下,不同浓度FK506对类神经细胞PC12细胞突起生长的影响,通过蛋白免疫印迹法检测FK506在常氧与缺氧条件下对PC12细胞热休克蛋白HSP70、缺氧诱导因子HIF-1α表达的影响,探讨FK506保护缺氧性神经元受损的可能性机制.结果表明:在缺氧及常氧状态下,1 1 000nmol·L-1浓度范围内FK506均可协同神经生长因子（NGF）促进PC12细胞突起的生长,其中,FK506 100nmol·L-1与NGF 10ng·mL-1对缺氧后PC12细胞突起生长的协同作用最为显著.该配比的FK506协同NGF对缺氧相关保护蛋白表达具有促进作用,其抗氧化损伤的作用可能与促进HSP70和HIF-1α的表达有关. Studies were performed to determine whether the drug（FK506）influenced the neuronal cells under the hypoxic conditions and the mechanism underlying.A model of neuronal hypoxia was constructed.The effect of FK506 and nerve growth factor（NGF）in hypoxic conditions promoting neurite growth were explored.Firstly the optimal concentration of FK506 and NGF were screened.In hypoxic conditions,the effects of different concentrations of FK506 on PC12（neural cell research model）cells neuritis outgrowth were quantitatively detected.The influence of FK506 and NGF on HSP70（heat shock protein 70）,HIF-1α（hypoxia inducible factor）protein expression of PC12 cells were detected by immunoblotting（Western blotting）.The preliminary mechanism of FK506 protect neurons under hypoxia were explored.The results show that PC12 cell neuritis outgrowth was collaboratively promoted by 1-1 000nmol·L-1 FK506 and 10ng·mL-1 NGF.Meanwhile,the neurite outgrowth of PC12 cells after hypoxia was most obvious after treated with FK506 100nmol·L-1 and NGF10ng·mL-1.The expression of HSP70 and HIF-1αprotein of PC12 cells was unregulated after treated with 100nmol·L-1 FK506 and 10ng·mL-1 NGF in hypoxia condition. 神经再生 神经生长因子（NGF） FK506 PC12细胞 HSP70 HIF-1α nerve regeneration nerve growth factor FK506 PC12cells HSP70 HIF-1α 10.14188/j.1671-8836.2015.04.016 Q421 国家自然科学基金青年基金（51403168）;国家科技重大专项973计划（2011CB606205）;湖北省自然科学基金（2013CFB354,2014CFB306）资助项目 2015-04-01 2021-03-31 4 61