鼻腔给药NMFGF1纳米粒改善血管性痴呆小鼠认知功能障碍

Nasal Administration of NMFGF1 Loaded Nanoparticles Improved Cognitive Dysfunction in Vascular Dementia Mice

目的考察鼻腔给药包载非促分裂活性酸性成纤维细胞生长因子(non-mitogenic acid fibroblast growth factor,NMFGF1)的纳米粒(NMFGF1-NPs)对于血管性痴呆(vascular dementia,VD)小鼠认知功能改善作用及其机制。方法应用水包水型乳化技术制备NMFGF1-NPs并进行质量表征。采用反复脑缺血再灌注法建立VD试验模型后分成假手术组、VD模型组、空白纳米粒组、NMFGF1溶液组及NMFGF1-NPs组,分别鼻腔给予相应形式药物干预。干预结束后,应用Morris水迷宫评价试验动物的学习及记忆功能,同时应用HE染色、FJB染色及Tunel凋亡染色等病理学方法评价试验动物海马神经元的形态、排列及凋亡指数(apoptosis index,AI),另外应用ELISA及Western blotting等分子生物学方法探讨鼻腔给药NMFGF1-NPs改善VD的分子机制。结果NMFGF1-NPs形态圆整,包封率(87.76±5.89)%。Morris水迷宫结果显示VD模型组小鼠的各项行为学指标均较假手术组有显著差异(P<0.01),同时病理结果显示,VD模型组小鼠海马神经元CA1区域神经元排列紊乱、细胞形态结构缺失且AI较假手术组显著增加(P<0.01),而经过鼻腔给药NMFGF1-NPs治疗组小鼠的各项行为学指标较VD模型组有显著改善,同时海马神经元细胞形态完整,排列整齐,AI指数较VD模型组及其他各个治疗干预组显著降低(P<0.01)。ELISA及Western blotting分析结果显示,鼻腔给药NMFGF1-NPs治疗组小鼠脑内MDA含量较VD模型组及其他各个治疗干预组显著下降(P<0.01),同时SOD,NO含量及Nrf2,SOD-1,GSTO1/2表达显著增加(P<0.01)。结论鼻腔给药NMFGF1-NPs给够通过激活Nrf2/ARE信号通路,发挥抗氧化应激损伤的作用,最终改善VD小鼠的学习及认知功能。

OBJECTIVE To investigate the effects of nasal administration of non-mitogenic acid fibroblast growth factor(NMFGF1)loaded nanoparticles(NMFGF1-NPs)on the improvement of cognitive function in vascular dementia(VD)mice and its mechanism.METHODS Nanoparticles containing NMFGF1(NMFGF1-NPs)were prepared by water-in-water emulsion technique and characterized.The mice were divided into sham group,VD model group,blank-NPs group,NMFGF1 solution group and NMFGF1-NPs group after repeated cerebral ischemia-reperfusion to establish VD test model,and then given the corresponding form of drug intervention by nasal cavity.After drug intervention,Morris water maze was used to evaluate the learning and memory function of the animals in each group from the perspective of behavior.Meanwhile,the morphology,arrangement and apoptosis index(AI)of hippocampal neurons in each group were evaluated by pathological methods such as HE staining,FJB staining and Tunel apoptosis staining.In addition,ELISA and Western blotting were used to investigate the molecular mechanism of NMFGF1-NPs improving VD by nasal administration.RESULTS The morphology of NMFGF1-NPs was round.The encapsulation rate of NMFGF1-NPs respectively was(87.76±5.89)%.Morris water maze results showed that the behavioral indexes of mice in VD model group were significantly different from those in sham operation group(P<0.01).At the same time,the pathological results showed that the neurons in the CA1 region of the hippocampus in the VD model group were disordered,the cells morphology and structure were missing,and the AI was significantly increased compared with that in the sham operation group(P<0.01).Meanwhile,compared with the VD model group,the NMFGF1-NPs treatment group showed significant improvement in various behavioral indexes,and the hippocampal neuron cells were intact and orderly,and the AI index was significantly decreased(P<0.01).ELISA and Western blotting analysis showed that compared with that of VD model group and other intervention groups,the content of MDA in the brain of NMFGF1-NPs treatment group was significantly decreased.While the content of SOD,NO and the expressions of Nrf2,SOD-1 and GSTO1/2 was significantly increased(P<0.01).CONCLUSION Nasal administration of NMFGF1-NPs can play the role of antioxidant stress damage by activating Nrf2/ARE signal pathway,and ultimately improve the learning and cognitive function of VD mice.