聚山梨酯80修饰的神经毒素纳米粒跨血脑屏障转运及细胞毒性  

Polysorbate-80 modified neurotoxin nanoparticle with its transport and cytotoxicity against blood-brain barrier

作　　者：赵燕敏[1] 夏爱晓[1] 魏颖慧[1] 阮叶萍[1] 李范珠[1] 

机构地区：[1]浙江中医药大学药学院,浙江杭州310053

出　　处：《药学学报》2010年第10期1312-1316,共5页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(30772793);浙江省自然科学基金资助项目(R207722)

摘　　要：研究聚山梨酯80修饰的神经毒素纳米粒(polysorbate-80modified neurotoxin nanoparticle,P-80-NT-NP)在血脑屏障(blood-brain barrier,BBB)体外模型上的通透性及细胞毒性。采用SD大鼠大脑分离培养脑微血管内皮细胞(rat brain microvascular endothelial cells,rBMECs)建立BBB体外模型,以神经毒素(neurotoxin,NT)组、空白纳米粒(nanoparticle,NP)组和未修饰的神经毒素纳米粒(neurotoxin nanoparticle,NT-NP)组作为对照,MTT法考察P-80-NT-NP对rBMECs的细胞毒性;采用Millicell insert培养rBMECs,荧光分光光度法测定P-80-NT-NP在基底侧中NT浓度,计算其表观通透系数。结果显示,当NT质量浓度≤200ng·mL-1时,NT组、NP组、NT-NP组和P-80-NT-NP组均无细胞毒性;NT质量浓度>200ng·mL-1时,NP组、NT-NP组和P-80-NT-NP组的细胞毒性显著大于NT组(P<0.01),且P-80-NT-NP组的细胞毒性较NP组和NT-NP组稍大,但无显著性差异(P>0.05)。当NT质量浓度为150ng·mL-1时,NT-NP组和P-80-NT-NP组的转运量均显著大于NT组(P<0.01),且P-80-NT-NP组的转运量较NT-NP组亦有显著增加(P<0.05),表明P-80修饰的NT纳米粒能跨BBB转运;但当P-80-NT-NP质量浓度>200ng·mL-1时,对rBMECs产生细胞毒性,为脑靶向纳米粒跨BBB转运的机制研究奠定基础。This study was aimed at the transport across blood-brain barrier （BBB） of polysorbate-80 modified neurotoxin loaded polybutylcyanoacrylate nanoparticle （P-80-NT-NP） and its cytotoxicity. An in vitro model of BBB using rat brain microvascular endothelial cells （rBMECs） was established. The cytotoxicity of P-80-NT-NP was measured by the MTT assays, where neurotoxin （hiT）, nanoparticle （NP）, neurotoxin nanoparticle （NT-NP） as control, and the permeability of P-80-NT-NP was determined by using of Millicell insert coculture with rBMECs and fluorescence spectrophotometry. MTT results showed that NT, NP, NT-NP and P-80-NT-NP were avirulent to rBMECs when the concentration of NT was lower than 200 ng·mL^-1. But the cytotoxicity of NP, NT-NP and P-80-NT-NP would be augmented accordingly as concentration increased （P 〈 0.01）, causing obvious reductions of cell survival rate, with no significant difference between them （P 〉 0.05）. When the concentration of NT was 150 ng·mL^-1, the permeability on rBMECs of P-80-NT-NP and NT-NP were both significantly higher than that of NT （P 〈 0.01）, and the permeability of P-80-NT-NP was greater than that of NT-NP （P 〈 0.05）. In conclusion, polysorbate-80 modified neurotoxin nanoparticles can transport across the BBB, while concentration of NT is greater than 200 ng·mL^-1, P-80-NT-NP has a little cytotoxicity against rBMECs.

关 键 词：聚山梨酯80 神经毒素纳米粒 血脑屏障 体外转运 细胞毒性 

分 类 号：R943[医药卫生—药剂学]