载睫状神经营养因子聚乙二醇-乳酸/羟基乙酸共聚物微泡联合超声对视网膜神经节细胞的保护作用  

Ciliary neurotrophic factor-loaded polyethylene glycol-poly lactic-co-glycolic acid microbubbles combined with ultrasound for protection of retinal ganglion cells

作　　者：王文婷[1] 刘苏[1] 王志刚[2] 周希瑗[1] 

机构地区：[1]重庆医科大学附属第二医院眼科,重庆400010 [2]重庆医科大学超声影像学研究所,重庆400010

出　　处：《中国医学影像技术》2013年第6期875-879,共5页Chinese Journal of Medical Imaging Technology

基　　金：国家自然科学基金重点项目(81130025);国家自然科学基金面上项目(81071157)

摘　　要：目的制备载睫状神经营养因子(CNTF)的聚乙二醇(PEG)-乳酸/羟基乙酸共聚物(PLGA)超声微泡造影剂(CNTF-PEG-PLGA),观察其联合超声对视神经损伤大鼠视网膜神经节细胞(RGCs)的保护作用。方法采用双乳化法制备CNTF-PEG-PLGA,并检测其基本特性。随机选取SD大鼠145只(双眼兼用),将其分为7组,采用视神经钳夹法制作大鼠视神经损伤模型。治疗后应用荧光金(FG)逆行标记法比较各组大鼠RGCs存活数;视网膜组织病理切片观察视网膜的形态结构改变,评价载药微泡及联合超声的安全性;生长相关蛋白-43(GAP-43)免疫组化染色,观察大鼠视网膜GAP-43的表达情况。结果CNTF-PEG-PLGA微泡平均粒径(312.5±57.35)nm,包封率62.35%,载药量0.298μg/mg,体外释放28天时微泡释放率达93.60%。FG标记RGCs示,在每个观察时间点G组平均RGCs计数均显著高于其他各损伤组(P<0.05),但仍低于A组(P<0.05);G组GAP-43表达可持续到伤后4周,且明显高于其他各组(P<0.05);视网膜组织病理切片示玻璃体腔注射微泡后视网膜未见明显炎性细胞浸润现象。结论载睫状神经营养因子PEG-PLGA微泡联合超声可增强药物对视神经损伤大鼠视网膜神经节细胞的保护作用,延长药物作用时间。Objective To prepare and observe the properties of ciliary neurotrophic factor-loaded polyethylene glycol-poly lactic-co-glycolic acid ultrasound contrast agents （CNTF-PEG-PLGA）, and to assess the protection effect of CNTF- loaded PEG-PLGA microbubbles combined with ultrasound on retinal ganglion cells （RGCs） after optic nerve injury in rats. Methods CNTF-loaded PEG-PLGA ultrasound microbubbles were prepared using double emulsion technique, and the properties of the ultrasound microbubbles were observed. Totally 145 adult SD rats were randomly divided into 7 groups. After optic nerve injury, RGCs were retrograde labeled with the fluorescent tracer fluorogold （FG） to count RGCs number. The expressions of growth associated protein 43 （GAP-43） were detected by immunohistochemistry on the 3rd day, the 1st, the 2nd and the 4th week after optic nerve injury. The retinal pathological morphology change was observed at different times. Results CNTF-loaded PEG-PLGA ultrasound microbubbles were smooth and spherical with a mean size of （312.5-57.35）nm, with the drug loading 0. 298 /~g/mg and encapsulation efficiency of 62.35~~. There were 93. 600//oo CNTF-PEG-PLGA release within 28 days. The results showed that after RGCs were retrograde labeled with FG, the RGCs count of group G was higher than that of other groups at every time, （all P〈0.05）, but less than group A （P〈0.05）. The expression of group Cr was higher than that of other groups （P〈0.05） on the 1st, the 2nd and the 4th week after optic nerve injury. PEC-PLGA microbubbles did not induce any histology signs of ocular inflammation after their injection in the vitreous. Conclusion The ciliary neurotrophic factor-loaded PEG-PLGA microbubbles combined with ultrasound can protect retinal ganglion cells against after optic nerve injury in rats.

关 键 词：聚乙二醇-乳酸 羟基乙酸共聚物 超声学 微泡 睫状神经营养因子 视网膜神经节细胞 

分 类 号：R774[医药卫生—眼科] R332[医药卫生—临床医学]