[PEAD-肝素-NGF]生物材料对糖尿病大鼠周围神经病变的改善作用  

Biomaterial of [PEAD-Heparin-NGF] Attenuates Diabetic Neuropathy in Rats

作　　者：李锐[1] 邹双[1] 刘彦隆[1] 张宏宇[1] 肖健[1] 

机构地区：[1]温州医科大学药学院浙江省生物技术制药工程重点实验室,浙江温州325035

出　　处：《中国药学杂志》2017年第11期941-947,共7页Chinese Pharmaceutical Journal

基　　金：国家自然科学资金资助项目(80213069)

摘　　要：目的探究新型生物材料poly(ethylene argininylaspartate diglyceride)(PEAD)-肝素-神经生长因子(PEAD-肝素-NGF)对糖尿病大鼠坐骨神经病变的作用。方法成年雄性SD大鼠腹腔注射链脲佐菌素制备糖尿病外周神经病变(DPN)模型,随机分为模型组、NGF治疗组及[PEAD-肝素-NGF]团聚体治疗组并给予对应的药物治疗,每周对各组进行行为学测试并在第30天将收集的坐骨神经进行HE染色及TUNEL凋亡检测。细胞水平上,将处于对数期的RSC 96大鼠雪旺细胞(SCs)培养于30 mmol·L^(-1)高糖环境下并给予NGF处理24 h,运用四甲基偶氮唑蓝(MTT)法和流式细胞术检测各组的凋亡情况。结果 DPN大鼠经[PEAD-肝素-NGF]团聚体治疗后,运动和感觉功能恢复良好,神经纤维节段性脱髓鞘情况改善明显并显著抑制SCs的凋亡。结论 [PEAD-肝素-NGF]团聚体治疗DPN效果显著,为该药应用于临床提供数据支持。OBJECTIVE To investigate the effects of [ PEAD-Heparin-NGF] coacervate on diabetic neuropathy in rats. METHODS Eight-week-old male SD rats （200 - 220 g） were intraperitoneal injection streptozotocin （STZ） at a does of 65 mg·kg-1 in phosphate-buffered saline （PBS） and rats with serum glucose concentrations of ≥16.7 mmol·L-1 were considered diabetic. All experimental rats had free access to rat chow and water. After 8 weeks, diabetic neuropathy rats were randomly divided into three groups： model group, free NGF group and NGF-coacervate group. Each group was administered corresponding therapeutic drugs to the right thigh and soleus muscles through a 1.0 mL syringe. The functional recovery of sensory and motor systems in all tested rats was assessed using walking track analysis and hot plate test at postin jection weeks 1, 2, 3 and 4. After 30 d, rats were anesthetized with 4% choral hydrate （10 mL·kg-1 ip） and perfused with with 0.9% NaCl. The nerves from both sides were dissected out and harvested for HE staining and TUNEL apoptosis asssy. For cell experiment, SCs were cultured in 30 mmol ·L-1 high glucose （HG） and treated with 100 ng·mL-1 NGF or NGF-coacervate which containing the same dosage NGF for 24 h. Cell viability and apoptotic analysis were determined by MTT assay and flow cytometer. RESULTS [ PEAD- Heparin-NGF] coacervate can effectively promote functional improvement of moter and sensory and significantly ameliorate neurological morphology as well as remarkably decrease cells apoptosis in the sciatic nerve of DPN rats. This therapeutical effect was also confirmed in cell level. CONCLUSION This new type of coacervate loaded with NGF shows striking effects on functional and morphometric recovery in diabetic neuropathy as well as SCs survival. The data from this study may have bearing on therapeutic strategies for improving diabetic neuropathy in clinical populations.

关 键 词：糖尿病周围神经病变 神经生长因子 雪旺细胞 高糖 团聚体 

分 类 号：R965[医药卫生—药理学]