神经生长因子纳米途径给药对糖尿病大鼠创面修复的研究

Experimental study on nerve growth factor with nanochannel administration for treating deep partial thickness scald in rats

目的:探讨局部应用神经生长因子-壳聚糖纳米颗粒缓释凝胶对1型糖尿病大鼠烫伤创面中血管内皮细胞生长因子(vascular endothelial growth factor, VEGF)、低氧诱导因子-1α(hypoxia inducible factor-1α, HIF-1α)、B淋巴细胞瘤-2基因(B-cell lymphoma-2, Bcl-2)及B淋巴细胞瘤相关X蛋白(Bcl-associated X protein, Bax)表达的影响及创面愈合的机制。方法:制作具有稳定性状的神经生长因子-壳聚糖纳米颗粒缓释凝胶并将其应用于1型糖尿病大鼠深Ⅱ度烫伤创面模型;将糖尿病大鼠模型分组,A组:神经生长因子治疗组;B组:壳聚糖治疗组;C组:神经生长因子-壳聚糖纳米颗粒缓释凝胶治疗组;D组:对照组。观察各组创面愈合率及VEGF、HIF-1α、Bcl-2及Bax等因子的表达。结果:制作出具有稳定性状的神经生长因子-壳聚糖纳米颗粒缓释凝胶,酶联免疫吸附法(enzyme-linked immunosorbent assay, ELISA)检测其封包率为28.29%,24 h后降解率为25.5%,无突释效应。通过创面用药干预,C组在第21天创面愈合率最高,而D组最低,与其他各组相比差异均有统计学意义(均P<0.05)。各组VEGF及Bcl-2的灰度值在第3~15天增长达到最高峰后开始回落,C组从第11天开始,D组从第7天开始,与其他各组相比差异均有统计学意义(均P<0.05),第21天C组的VEGF和Bcl-2表达最高,两者在D组表达最低。HIF-1α和Bax灰度值均从第3天的最高峰开始回落,C组降速最快,D组降速最慢,第21天C组的HIF-1α和Bax灰度值最低,两者在D组最高,与其他各组间差异均有统计学意义(均P<0.05)。结论:该研究能制作出性状稳定的神经生长因子-壳聚糖纳米颗粒缓释凝胶制剂,其缓释给药途径能对糖尿病大鼠慢性创面的修复起到加速作用。

Objective: To investigate the effects of local application of nerve growth factor(NGF)-chitosan nanoparticle sustained-release gel on the mechanism of wound healing with Ⅱ degree deep scald and the expression of vascular endothelial growth factor(VEGF), hypoxia inducible factor-1α(HIF-1α), B-cell lymphoma-2(Bcl-2) and Bcl-associated X protein(Bax) in type 1 diabetic rats. Methods: The stable NGF-chitosan nanoparticle sustained release gel was constructed for treatment of Ⅱdegree deep scald wound. A total of 60 type 1 diabetic rats were enrolled and randomized equally to receive NGF(Group A),chitosan(Group B), NGF-chitosan nanoparticle sustained release gel(Group C) and no treatment(Group D). The wound area,the rate of wound healing and expressions of VEGF, HIF-1α, Bcl-2 and Bax were determined on 3, 7, 15, and 21 day post scalding days(PSDs). Results: The stable NGF-chitosan nanoparticle sustained release gel was successfully constructed, with the packet rate of 28.29% by enzyme-linked immunosorbent assay(ELISA), the degradation rate in 24 hours of 25.5% and no burst release effect. The rate of wound healing in day 21 was the highest in the group C and lowest in the group D,statistically different from other groups(P<0.05). The expression of VEGF and Bcl-2 peaked in day 3-15 among groups and began to be statistically different from other groups in day 11 for group C and in day 7 for group D. The expressions in day 21 were highest in group C and lowest in group D. The expressions of HIF-1α and Bax peaked in day 3 and showed the fastest falling velocity in group C and the lowest falling velocity in group D. The expressions of HIF-1α and Bax in day 21 was lowest in group C and highest in group D, statistically different from other groups( P<0.05). Conclusion: This study can successfully construct a trait-stable NGF-chitosan nanoparticle sustained-release gel and the sustained-release administration route can accelerate the repair of chronic wounds in diabetic rats.