Pdots-RVG-Curcumin纳米复合物穿透体外血脑屏障模型的研究  

Penetration of Pdots-RVG-Curcumin Nanocomposite Through Blood Brain Barrier Model in Vitro

作　　者：文欣宜 苏炳银[1,3] 李淑蓉[1,2] 韩玉萍 Wen Xinyi;Su Bingyin;Li Shurong;Han Yuping(Key Laboratory of Development and Regeneration of Sichuan Province,Chengdu Medical College,Chengdu 610500,China;Department of Pathology and Pathophysiology,Chengdu Medical College,Chengdu 610500,China;Department of Histology and Embryology,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院发育与再生四川省重点实验室,成都610500 [2]成都医学院病理学与病理生理学教研室,成都610500 [3]成都医学院组织胚胎学教研室,成都610500

出　　处：《成都医学院学报》2021年第3期278-284,共7页Journal of Chengdu Medical College

基　　金：四川省科技厅应用基础研究重点项目(No:2019YJ0366,No:21YYJC0342);中国科协青年人才托举项目(No:2019QNRC001);发育与再生四川省重点实验室研究基金重点项目(No:SYS17-006)。

摘　　要：目的探究Pdots-RVG-Curcumin纳米复合物穿透血脑屏障并靶向多巴胺能神经元的作用。方法采用纳米共沉淀的方法制备半导体聚合物量子点(Pdots)。利用静电吸附原理,将其连接狂犬病毒糖蛋白(RVG)以及姜黄素(Cur),制备Pdots-RVG-Cur复合物;透射电子显微镜观察Pdots、Pdots-RVG、Pdots-RVG-Cur的形态,动态光散射技术测量三者的粒径,并检测其稳定性;CCK-8检测Pdots-RVG的生物相容性;利用b.End3细胞和带有PET膜的细胞小室,构建体外血脑屏障(BBB)模型,通过4 h试漏实验和荧光素钠通透性实验评价模型的功能、状态;将BBB体外模型移至接种MN9D细胞的培养皿之上,将游离Cur和Pdots-RVG-Cur纳米复合物加入到上层BBB模型中,激光共聚焦显微镜(CLSM)观察MN9D细胞对两种药物的摄取情况,从而确定其穿透BBB的能力。结果成功构建Pdots-RVG-Cur纳米复合物,呈球形,粒径为320 nm左右;CCK-8测得当Pdots浓度在0~50 mg/L范围内,Pdots-RVG表现出良好的生物相容性;成功构建BBB体外模型,CLSM下观察到Pdots-RVG-Cur组比Cur组进入MN9D中Cur的量更多。结论本项目合成的Pdots-RVG-Cur纳米复合物给药体系提高Cur穿透体外BBB以及被神经元摄取的能力,增强Cur在脑部疾病的应用。Objective To investigate whether Pdots-RVG-Curcumin nanocomposite can penetrate the blood brain barrier(BBB)and target dopaminergic neurons.Methods Semiconducting polymer quantum dots(Pdots)were prepared by nano coprecipitation method,which were connected with rabies virus glycoprotein(RVG)and curcumin(Cur)according to the principle of electrostatic adsorption to prepare Pdots-RVG-Cur nanocomposite.The morphologies of Pdots,Pdots-RVG and Pdots-RVG-Cur were observed by transmission electron microscope.And dynamic light scattering technology was used to measure the particle size of the three and detect their stability.Cell counting kit-8(CCK-8)was used to detect the biocompatibility of Pdots-RVG.The BBB model in vitro was established by using b.End3 cells and cell chambers with PET membrane.The function and status of the model were evaluated by 4 h leakage test and fluorescein sodium permeability test.The BBB model in vitro was transferred to a culture dish inoculated with MN9D cells.And the free Cur and Pdots-RVG-Cur nanocomposites were added to the upper BBB model.The uptake of the two drugs by MN9D cells was observed by confocal laser scanning microscopy(CLSM)to determine the ability of MN9D cells to penetrate BBB.Results The Pdots-RVG-Cur nanocomposite was successfully constructed,which was spherical with a particle size of about 320 nm.CKK-8 measured that when the Pdots concentration was in the range of 0-50 mg/L,Pdots-RVG showed good biocompatibility.The BBB in vitro model was successfully constructed.Under CLSM,it was observed that the amount of Cur entering MN9D cells in the Pdots-RVG-Cur group was more than that in the Cur group.Conclusion The drug delivery system of Pdots-RVG-Cur nanocomposite synthesized in this project improves the possibility of Cur to penetrate BBB in vitro and be taken up by neurons,which enhances the application of Cur in brain diseases.

关 键 词：血脑屏障 姜黄素 纳米载体 半导体聚合物量子点 

分 类 号：R971[医药卫生—药品]