神经生长因子温敏凝胶对大鼠坐骨神经损伤的修复作用
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摘要
目的:制备神经生长因子(NGF)温度敏感型原位凝胶,以大鼠坐骨神经损伤为模型,探讨NGF温敏凝胶对大鼠坐骨神经损伤的修复作用。方法:制备NGF温敏凝胶,利用星点设计-效应面法优化其处方。将50只大鼠随机分为正常组,模型组,NGF注射剂组(10 mg·L~(-1)),NGF低剂量温敏凝胶组(10 mg·L~(-1))和NGF高剂量温敏凝胶组(20 mg·L~(-1)),建立大鼠坐骨神经损伤模型;以大鼠行为学,坐骨神经功能指数(SFI),撤回反射时间的测定实验,腓肠肌湿重比的测定以及组织形态学变化为指标,综合考察NGF温敏凝胶对损伤坐骨神经的影响。结果:处方优化后NGF温敏凝胶的胶凝温度35. 2℃,符合注射用标准。术后4~8周,各时间点测得NGF高剂量温敏凝胶组大鼠的SFI和腓肠肌湿重比都显著高于模型组和NGF注射剂组,但其撤回反射时间显著低于模型组和NGF注射剂组,且作用呈剂量依赖性。由苏木精-伊红(HE)染色试验可知,NGF高剂量温敏凝胶组的大鼠坐骨神经损伤区再生神经纤维排列较模型组整齐、密集,而且连续性也较好。结论:NGF温敏凝胶能促进大鼠坐骨神经损伤的修复。
Objective: To prepare nerve growth factor( NGF) temperature sensitive in situ gel and investigate its therapeutic effect on sciatic nerve injury of rats. Method: NGF thermosensitive gel was prepared and its prescription was optimized by central composite design-response surface methodology. Fifty rats were randomly divided into the normal group,model group,NGF injection group( 10 mg·L~(-1)),NGF low-dose( 10 mg·L~(-1))and high-dose( 20 mg·L~(-1)) thermosensitive gel groups,and sciatic nerve injury model of rats was established.The effect of NGF thermosensitive gel on the injury of sciatic nerve were comprehensively examined by taking rat behavior,sciatic nerve function index( SFI), time of withdrawal reflex, wet weight ratio of gastrocnemius muscle,and histomorphological changes as indicators. Result: The gelation temperature of NGF thermosensitive gel was 35. 2 ℃ after the formulation being optimized,which was in line with the standard for injection. Foureight weeks after operation, the SFI and wet weight ratio of gastrocnemius muscle in rats of NGF high-dose thermosensitive gel group were significantly higher than those in the model group and NGF injection group,but its time of withdrawal reflex was significantly lower than those in the model group and NGF injection group,and the effect was in a dose-dependent manner. Arrangement of regenerated nerve fibers in sciatic nerve injury area of rats from NGF high-dose thermosensitive gel group was more tidy,dense and continuous than that of the model group.Conclusion: NGF thermosensitive gel can promote repair of sciatic nerve injury in rats.
Objective: To prepare nerve growth factor( NGF) temperature sensitive in situ gel and investigate its therapeutic effect on sciatic nerve injury of rats. Method: NGF thermosensitive gel was prepared and its prescription was optimized by central composite design-response surface methodology. Fifty rats were randomly divided into the normal group,model group,NGF injection group( 10 mg·L~(-1)),NGF low-dose( 10 mg·L~(-1))and high-dose( 20 mg·L~(-1)) thermosensitive gel groups,and sciatic nerve injury model of rats was established.The effect of NGF thermosensitive gel on the injury of sciatic nerve were comprehensively examined by taking rat behavior,sciatic nerve function index( SFI), time of withdrawal reflex, wet weight ratio of gastrocnemius muscle,and histomorphological changes as indicators. Result: The gelation temperature of NGF thermosensitive gel was 35. 2 ℃ after the formulation being optimized,which was in line with the standard for injection. Foureight weeks after operation, the SFI and wet weight ratio of gastrocnemius muscle in rats of NGF high-dose thermosensitive gel group were significantly higher than those in the model group and NGF injection group,but its time of withdrawal reflex was significantly lower than those in the model group and NGF injection group,and the effect was in a dose-dependent manner. Arrangement of regenerated nerve fibers in sciatic nerve injury area of rats from NGF high-dose thermosensitive gel group was more tidy,dense and continuous than that of the model group.Conclusion: NGF thermosensitive gel can promote repair of sciatic nerve injury in rats.
引文
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[12]顼佳音,熊欣,陈燕军.可注射参麦温度敏感型原位凝胶的制剂学评价[J].中国实验方剂学杂志,2012,18(13):60-64.
[13]王锐,殷悦,曲炳楠,等.蛇床子素微囊-温敏凝胶的制备与质量分析研究[J].中国药学杂志,2017,52(12):1044-1048.
[14]黄嘉元,曾优美,杨佩佩,等.马来酸噻吗洛尔眼用温敏凝胶的处方筛选与体外释放评价[J].中国医院药学杂志,2015,35(24):2196-2200.
[15]张越,瞿叶清,陈军,等.阿魏酸脂质体的体外释放度考察[J].中国实验方剂学杂志,2016,22(19):1-5.
[16]张蕾,芮永军,王骏.足迹分析法评定超短波促进大鼠坐骨神经损伤后功能恢复的研究[J].交通医学,2007,21(5):485-486.
[17]王维,苑秀华,王中莉,等.坐骨神经损伤模型大鼠神经传导速度及损伤运动神经元内生长相关蛋白43表达与磁刺激干预[J].中国组织工程研究,2011,15(46):8617-8620.
[2]赵广翊,丁旭东,黄威,等.神经生长因子预处理对神经损伤大鼠脊髓脱髓鞘反应的影响[J].中国医科大学学报,2016,45(9):783-786,792.
[3]李明明,李林林,季晓燕.神经生长因子治疗周围神经损伤的疗效观察[J].中国实用神经疾病杂志,2015,18(4):24-26.
[4] Tucker K L,Meyer M,Barde Y A. Neurotrophins are required for nerve growth during development[J]. Nat Neurosci,2001,4(1):29-37.
[5]徐莉.神经生长因子的研究进展[J].中国生物制品学杂志,2014,27(1):131-134.
[6]陈庆真,施明祥,刘盛飞,等.鼠神经生长因子不同给药方式修复周围神经损伤[J].中国组织工程研究,2014,18(33):5356-5360.
[7] Cattaneo A,Calissano P. Nerve growth factor and Alzheimer's disease:new facts for an old hypothesis[J].Mol Neurobiol,2012,46(3):588-604.
[8] Lambiase A,Sacchetti M,Bonini S. Nerve growth factor therapy for corneal disease[J]. Curr Opin Ophthalmol,2012,23(4):296-302.
[9] Bronzetti E,Artico M,Lo V V,et al. Expression of neurotransmitters and neurotrophins in human adenoid tissue[J]. Int J Mol Med,2005,15(6):921-928.
[10]张贝贝.可注射a FGF/磺化壳寡糖温敏水凝胶的制备及药效学研究[D].广州:暨南大学,2016.
[11]梁浩明,龙晓英,卢彬.黄芩苷外用温敏凝胶的处方设计及优化[J].中国实验方剂学杂志,2014,20(24):34-38.
[12]顼佳音,熊欣,陈燕军.可注射参麦温度敏感型原位凝胶的制剂学评价[J].中国实验方剂学杂志,2012,18(13):60-64.
[13]王锐,殷悦,曲炳楠,等.蛇床子素微囊-温敏凝胶的制备与质量分析研究[J].中国药学杂志,2017,52(12):1044-1048.
[14]黄嘉元,曾优美,杨佩佩,等.马来酸噻吗洛尔眼用温敏凝胶的处方筛选与体外释放评价[J].中国医院药学杂志,2015,35(24):2196-2200.
[15]张越,瞿叶清,陈军,等.阿魏酸脂质体的体外释放度考察[J].中国实验方剂学杂志,2016,22(19):1-5.
[16]张蕾,芮永军,王骏.足迹分析法评定超短波促进大鼠坐骨神经损伤后功能恢复的研究[J].交通医学,2007,21(5):485-486.
[17]王维,苑秀华,王中莉,等.坐骨神经损伤模型大鼠神经传导速度及损伤运动神经元内生长相关蛋白43表达与磁刺激干预[J].中国组织工程研究,2011,15(46):8617-8620.