PNV@LIR纳米囊泡的制备及其细胞功能验证

PNV@LIR preparation of nanovesicles and validation of their cellular functions

利用胞吞法制备尺寸均匀的血小板包裹利拉鲁肽的纳米囊泡。从全血中经过离心、重悬和洗涤得到大小均匀的血小板纳米囊泡,进一步将利拉鲁肽与血小板纳米囊泡37℃恒温摇床共孵育4 h,得到血小板载利拉鲁肽纳米囊泡(PNV@LIR)。检测其理化和功能指标:PNV@LIR纳米囊泡的颗粒大小、微观形貌、Zeta电位、表面蛋白受体、细胞毒性和抗氧化损伤能力。结果显示,制备得到的PNV@LIR为稳定的140 nm的囊泡状,PNV@LIR的Zeta电位值为(-23.2±1.85)mV。SDS-PAGE分析证实了PNV@LIR表面的膜蛋白结构的存在。体外细胞实验显示:PNV@LIR纳米囊泡生物相容性良好,且具有优异的抗氧化损伤能力。研究应用离心和共孵育相结合的方法成功制备出大小均一的PNV@LIR纳米囊泡,SDS-PAGE结果表明其表面保留了血小板本身的蛋白受体;体外细胞实验表明PNV@LIR纳米囊泡无细胞毒性,且具有明显的抗氧化应激损伤能力。

The purpose of this article is to prepare platelet coated liraglutide nanovesicles with uniform size by endocytosis.Platelet nanovesicles of uniform size were obtained from whole blood by centrifugation,resuspension and washing.Liraglutide was further incubated with platelet nanovesicles for 4 h at 37℃shaking table.Then platelet-loaded liraglutide nanocapsules(PNV@LIR)were obtained.The physicochemical and functional indexes of the nanoparticles were detected including the particle size,microstructure,Zeta potential,surface protein receptors,cytotoxicity and antioxidant damage ability of PNV@LIR.The results show that the obtained PNV@LIR was a stable 140 nm vesicle shape,and the Zeta potential value of PNV@LIR was(-23.2±1.85)mV.SDS-PAGE was used to verify the existence of membrane protein structure on the surface of PNV@LIR.In vitro cell experiments showed that PNV@LIR nanovesicles had good biocompatibility and excellent anti-oxidative damage ability.Therefore,homogeneous PNV@LIR nanoveses were successfully prepared by a combination of centrifugation and co-incubation.The surface of the nanoveses retained the platelet protein receptors.In vitro cell experiments showed that PNV@LIR vesicles were non-cytotoxic and had obvious anti-oxidative damage ability.