新型温敏型肝素-泊洛沙姆水凝胶包载神经生长因子对糖尿病外周神经损伤修复的作用

李锐, 李多慧, 全大萍, 肖健

中国药学杂志 ›› 2019, Vol. 54 ›› Issue (12) : 992-999.

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中国药学杂志
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中国药学杂志 ›› 2019, Vol. 54 ›› Issue (12) : 992-999. DOI: 10.11669/cpj.2019.12.012
论著

新型温敏型肝素-泊洛沙姆水凝胶包载神经生长因子对糖尿病外周神经损伤修复的作用

  • 李锐1,2, 李多慧2, 全大萍1, 肖健2*
作者信息 +

Novel Thermosensitive Heparin-poloxamer Hydrogel Bridge NGF to Treat Peripheral Nerve Injury in Diabetics Rats

  • LI Rui1,2, LI Duo-hui2, QUAN Da-ping1, XIAO Jian2*
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文章历史 +

摘要

目的 探讨神经生长因子-肝素泊洛沙姆水凝胶(NGF-HP)对糖尿病大鼠坐骨神经损伤的修复作用。方法 成年雄性 SD大鼠腹腔注射链脲佐菌素以制备糖尿病(DM)模型。成模后,暴露并分离术侧的坐骨神经,运用静脉夹压迫神经以造成外周神经损伤(PNI)。同时,将其随机分为模型组(PNI-diabetics),游离NGF组(free NGF)及NGF-HP水凝胶组(NGF-HP hydrogel)。每组给予对应的药物治疗,同时在神经损伤后的每周进行斜板试验和BBB评分。30 d后,处死各组大鼠并收集术侧的坐骨神经,运用免疫印迹检测各组神经内部结构和功能蛋白的表达,运用免疫荧光、Masson三色染色和透射电镜观察各组神经内轴突和髓鞘的再生。结果 该水凝胶可装载一定剂量的神经生长因子(NGF)并缓控其释放。体内实验结果表明,单次注射一定体积的NGF-HP水凝胶后,PNI-diabetics大鼠运动功能恢复良好,神经内GAP-43、MAP-2、MBP及S-100的含量出现显著地上调,神经纤维的再生效果良好且脱髓鞘情况改善显著。结论 NGF-HP对PNI-diabetics疾病具有优越的疗效,为该药应进入临床提供理论依据。

Abstract

OBJECTIVE To investigate the effects of NGF-HP thermosensitive hydrogel on facilitating structural and functional regeneration in diabetic rats with sciatic nerve crush injury. METHODS Eight-week-old male SD rats (200-220 g) were intraperitoneally injected wither steptozocin(STZ) to induce diabetes. After success of model establishment, the sciatic nerve of the diabetes rats were made crushed through two vascular clips force. Muscle and skin were then closed with 5-0 stitches. Following surgery, the rats were randomly divided into three groups: PNI-diabetics group, free NGF group and NGF-HP hygrogel group. Each group received corresponding therapeutic drugs through a microsyringe. The motor recovery in all tested rats was assessed using Basso-Beattie-Bresnahan (BBB) locomotion scale and inclined plane test at indicated time points. After 30 d, rats were sacrificed, the crushed nerve and corresponding gastrocnemius muscle were harvested and the pathology index was assessed.The expressions of structural and functional proteins were detected through immunoblotting.The improvement of axon and myelination regeneration were evaluated via immunofluorescence, Masson′s trichrom stain and transmission electron microscope. RESULTS NGF-HP not only had a good affinity for a certain amounts of nerve growth factor (NGF), but also controlled its release in a steady fashion in vitro. In vivo, compared with administration of direct free NGF, single injection of NGF-HP hydrogel was more effective at upregulating the expression of nerve associated structural and functional proteins, enhancing axonal regeneration and remyelination, as well as improving motor function recovery. CONCLUSION This new type of hydrogel loaded with NGF shows striking effects on functional and morphometric recovery on peripheral nerve injury (PNI) following diabetes,which may provide a theoretical basis strategies for remedying PNI-diabetes in clinical populations.

关键词

外周神经损伤 / 糖尿病 / 肝素-泊洛沙姆水凝胶 / 神经生长因子 / 再髓鞘化

Key words

peripheral nerve injury / diabetes / heparin-poloxamer hydrogel / nerve growth factor / remyelination

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李锐, 李多慧, 全大萍, 肖健. 新型温敏型肝素-泊洛沙姆水凝胶包载神经生长因子对糖尿病外周神经损伤修复的作用[J]. 中国药学杂志, 2019, 54(12): 992-999 https://doi.org/10.11669/cpj.2019.12.012
LI Rui, LI Duo-hui, QUAN Da-ping, XIAO Jian. Novel Thermosensitive Heparin-poloxamer Hydrogel Bridge NGF to Treat Peripheral Nerve Injury in Diabetics Rats[J]. Chinese Pharmaceutical Journal, 2019, 54(12): 992-999 https://doi.org/10.11669/cpj.2019.12.012
中图分类号： R965   

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基金

国家自然资金项目资助(81802238)

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