NGF缓释系统促进大鼠坐骨神经电损伤后再生的研究
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摘要
研究背景与总体思路
现已证实，神经生长因子(nerve growth factor，NGF)具有明确的促进周围神经损伤后再生的作用，但在实际应用中，由于NGF活性半衰期较短，在水溶液中易失活，且易受湿度、酸碱度等多种因素的影响，因而限制了其临床应用。为此，寻找合理有效的给药途径及制剂已成为当务之急。
近年来神经电损伤发病率越来越高，其损伤机制较复杂，目前临床无合适有效的治疗方法，多为保守疗法，愈后较差，因而采取积极的治疗手段介入，促进受损神经尽快修复，缩短其功能恢复时间，将具有重要意义。
为此，我们设想：制备一种NGF缓释系统，这种系统能延缓释放NGF，且释放的NGF具有生物活性，将其应用于周围神经电损伤段局部，使其在局部缓慢释放活性NGF，可能在神经再生过程中维持较长时间的作用，促进周围神经再生。
据此，本工作的总体研究思路为：
首先，制备β-NGF缓释系统，用ELISA、DRG培养技术验证该系统能否延缓释放β-NGF，释放的β-NGF是否具有生物活性，并检测该系统中各种成分的作用和必要性。
其次，设计并制备一种稳定的能尽量模拟临床以“真性电损伤”为主的坐骨神经电损伤动物模型，通过光镜、电镜、坐骨神经功能指数、运动神经传导速度等观察其相应的形态、功能学改变，客观评价神经电损伤后的相应变化及再生能力，探讨其影响神经再生能力的可能机制，为下一步的实验研究提供参考。
最后，将β-NGF缓释系统应用于大鼠坐骨神经电损伤段局部，应用
Background and Research Strategy
It was conclusively confirmed that nerve growth factor could enhance peripheral nerve regeneration after injury. However, due to the short half-life action of NGF and suffered deactivation easily from humidity and acidity or alkalinity, the clinical application is difficult consequently. It is necessary to search a new reasonable way of drug administration therefore.
The incidence rate of nerve electrical burn is higher recently. There has no efficient therapy scheme in clinic at present. It is important to search an effective way to heal the impaired nerve as soon as possible to shorten the recovery time.
Thus, our experiment protocol is that to develop a NGF delivery system for use in repair of peripheral nerve after electric injury and we hypothesize that it would provide localized release of NGF in an active way to enhance peripheral nerve regeneration.
Based on above hypothesis, we conduce the researches as follows:
Firstly, we prepared a NGF controlled release system. We evaluated the ability of the delivery system to prolong the release of NGF in absence of cells by Sandwich ELISA and we determined if the delivery system could delivery bioactive NGF in a controlled manner and evaluated the necessity of the composition by DRG culture model.
Then we design an animal experiment model of peripheral nerve electric injury which mimicry clinical electric burn. By using light microscope and electron microscope to observe the changes of nerve morph and using SFI and NCV to observe the changes
现已证实，神经生长因子(nerve growth factor，NGF)具有明确的促进周围神经损伤后再生的作用，但在实际应用中，由于NGF活性半衰期较短，在水溶液中易失活，且易受湿度、酸碱度等多种因素的影响，因而限制了其临床应用。为此，寻找合理有效的给药途径及制剂已成为当务之急。
近年来神经电损伤发病率越来越高，其损伤机制较复杂，目前临床无合适有效的治疗方法，多为保守疗法，愈后较差，因而采取积极的治疗手段介入，促进受损神经尽快修复，缩短其功能恢复时间，将具有重要意义。
为此，我们设想：制备一种NGF缓释系统，这种系统能延缓释放NGF，且释放的NGF具有生物活性，将其应用于周围神经电损伤段局部，使其在局部缓慢释放活性NGF，可能在神经再生过程中维持较长时间的作用，促进周围神经再生。
据此，本工作的总体研究思路为：
首先，制备β-NGF缓释系统，用ELISA、DRG培养技术验证该系统能否延缓释放β-NGF，释放的β-NGF是否具有生物活性，并检测该系统中各种成分的作用和必要性。
其次，设计并制备一种稳定的能尽量模拟临床以“真性电损伤”为主的坐骨神经电损伤动物模型，通过光镜、电镜、坐骨神经功能指数、运动神经传导速度等观察其相应的形态、功能学改变，客观评价神经电损伤后的相应变化及再生能力，探讨其影响神经再生能力的可能机制，为下一步的实验研究提供参考。
最后，将β-NGF缓释系统应用于大鼠坐骨神经电损伤段局部，应用
Background and Research Strategy
It was conclusively confirmed that nerve growth factor could enhance peripheral nerve regeneration after injury. However, due to the short half-life action of NGF and suffered deactivation easily from humidity and acidity or alkalinity, the clinical application is difficult consequently. It is necessary to search a new reasonable way of drug administration therefore.
The incidence rate of nerve electrical burn is higher recently. There has no efficient therapy scheme in clinic at present. It is important to search an effective way to heal the impaired nerve as soon as possible to shorten the recovery time.
Thus, our experiment protocol is that to develop a NGF delivery system for use in repair of peripheral nerve after electric injury and we hypothesize that it would provide localized release of NGF in an active way to enhance peripheral nerve regeneration.
Based on above hypothesis, we conduce the researches as follows:
Firstly, we prepared a NGF controlled release system. We evaluated the ability of the delivery system to prolong the release of NGF in absence of cells by Sandwich ELISA and we determined if the delivery system could delivery bioactive NGF in a controlled manner and evaluated the necessity of the composition by DRG culture model.
Then we design an animal experiment model of peripheral nerve electric injury which mimicry clinical electric burn. By using light microscope and electron microscope to observe the changes of nerve morph and using SFI and NCV to observe the changes
引文
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