男性避孕纳米粒的制备与体外实验研究  

Preparation and in vitro effect of FSHR_(32-44)/ PLGA nanoparticles as male contraceptive

作　　者：徐萍萍[1] 杨利华[1,2] 魏桂芝 张建祥[3] 东亚君[1] 梁志清[1] 

机构地区：[1]第三军医大学西南医院妇产科,重庆400038 [2]唐山市工人医院妇产科,河北唐山063000 [3]第三军医大学药学院药剂教研室,重庆400038

出　　处：《第三军医大学学报》2015年第4期351-355,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81270731)~~

摘　　要：目的构建聚乳酸-羟乙酸共聚物(polylactic acid-glycolic acid,PLGA)包载促卵泡生成素受体表位肽(FSHR32-44)的纳米粒,探讨PLGA材料作为避孕疫苗载体的可行性。方法使用PLGA为载体,制备出包载促卵泡生成素受体的B细胞表位(FSHR32-44)的纳米粒,透射电镜(TEM)观察表观形态,粒径仪检测粒径分布范围和Zeta电位,体外释放试验检测其理化特性。激光共聚焦显微镜观察小鼠骨髓来源树突状细胞(DCs)吞噬FSHR可溶性多肽和FSHR/PLGA NP的能力,流式细胞仪检测FSHR、PLGA NP和FSHR/PLGA NP促骨髓来源DCs成熟能力。结果单乳-溶剂挥发法制备的FSHR/PLGA NP大小粒径分布均匀、表面规整,粒径(296.3±3.5)nm,Zeta电位(-16.1±0.4)m V,FSHR包封率(62.71±2.83)%,并具有缓慢释放抗原的能力;体外细胞水平实验结果显示FSHR表位肽纳米粒易于被DCs摄取,有剂量和时间依赖性,能显著上调DCs表面CD40、CD86、CD83和MHC-Ⅱ功能分子的表达。结论 PLGA包载FSHR多肽有良好的包封率,易被DCs吞噬,并有强的促进DCs成熟作用,可以作为FSHR32-44避孕疫苗的载体。Objective To synthesize nanoparticles（ NP） with follicle-stimulating hormone receptor peptide（ FSHR32- 44） encapsulated by poly（ lactic acid-glycolic acid）（ PLGA）,which is biodegradable polymer material approved by Food and Drug Administration,and explore whether PLGA carrier is suitable for contraceptive vaccine. Methods Emulsification process was used for preparation of FSHR32- 44-encapsulated PLGA NP（ FSHR32- 44/ PLGA NP）, and the morphology was observed with a transmission electron microscope. The particle size distribution and Zeta potential were detected with a laser potential particle size instrument,and the in vitro releasing characteristic of the NP was studied. A laser scanning confocal microscope was employed to observe the uptake capacity of bone marrow-derived dendritic cells（ BMDCs） to soluble FSHR32- 44 and FSHR32- 44/ PLGA NP,and a flow cytometer was used to detect the phenotypic changes of BMDCs co-cultured with FSHR32- 44,PLGA NP and FSHR32- 44/ PLGA NP,respectively. Results FSHR32- 44/ PLGA NP had uniform particle size distribution and regular surface,and could slowly release antigens,with average particle size of 296. 3 ± 3. 5 nm,Zeta potential of- 16. 1 ± 0. 4 m V,FSHR32- 44 encapsulation efficiency of 62. 71% ± 2. 83%,and total release rate of 98. 3%. The in vitro cell test showed that FSHR32- 44 was easily up-taken by BMDCs in a dose- and time-dependent manner,and the expressions of CD40,CD86, CD83 and MHC-Ⅱ on the surface of BMDCs were up-regulated. Conclusion The FSHR32- 44/ PLGA NP have good encapsulation efficiency,and can be easily up-taken by BMDCs to promote maturation of BMDCs. PLGA can be used as the carrier of FSHR32- 44 contraceptive vaccine.

关 键 词：FSHR PLGA 纳米疫苗 避孕 

分 类 号：R943[医药卫生—药剂学] R965[医药卫生—药学]