CAJ | 학술논문

目的探讨乳酸/羟基乙酸共聚物(PLGA)装载的促红细胞生成素(EPO)缓释微球(EPO-PLGA微球)经玻璃体腔注射对大鼠视神经挫伤模型中受损视网膜神经节细胞(RGC)的保护作用.方法选取成年SD大鼠,建立视神经挫伤模型.建模后分别经玻璃体腔内注射含10 IU EPO的PLGA微球(EPO-PLGA组)、10 IU EPO(EPO组)、5 μl空白PLGA(PLGA组)、5 μl PBS(PBS组),另设未治疗组不予玻璃体腔注药.术后5 d和2周,做视网膜切片,对各组RGC凋亡情况行TUNEL检测;术后23 d,DiI上丘逆标RGC,并于术后4周处死大鼠,视网膜铺片观察各组RGC存活情况;每组各个时间点分别处死6只SD大鼠.采用方差分析对结果进行比较.结果 TUNEL检测显示,术后5 d和2周,各组均可见TUNEL阳性细胞,其中EPO-PLGA组和EPO组TUNEL阳性细胞显著减少,其细胞凋亡率明显少于PLGA组、PBS组及未治疗组.术后4周,视网膜铺片RGC计数显示,正常SD大鼠RGC密度为(2387.7±164.9)个/mm2,未治疗组为(748.3±58.8)个/mm2,EPO-PLGA组为(1296.7±157.6)个/mm2,EPO组为(1418.5±154.9)个/mm2,PLGA组为(821.7±52.1)个/mm2,PBS组为(804.4±86.4)个/mm2;可见EPO-PLGA组和EPO组较未治疗组细胞密度显著增高,具有明显的RGC保护作用(P均＜0.01),而EPO-PLGA组和EPO组间差异无统计学意义(P=0.065).结论 EPO-PLGA缓释微球与EPO具有等效的RGC保护作用,这为进一步观察EPO-PLGA缓释微球的长效神经保护作用奠定了基础.
목적 탐토유산/간기을산공취물(PLGA)장재적촉홍세포생성소(EPO)완석미구(EPO-PLGA미구)경파리체강주사대대서시신경좌상모형중수손시망막신경절세포(RGC)적보호작용.방법 선취성년SD대서,건립시신경좌상모형.건모후분별경파리체강내주사함10 IU EPO적PLGA미구(EPO-PLGA조)、10 IU EPO(EPO조)、5 μl공백PLGA(PLGA조)、5 μl PBS(PBS조),령설미치료조불여파리체강주약.술후5 d화2주,주시망막절편,대각조RGC조망정황행TUNEL검측;술후23 d,DiI상구역표RGC,병우술후4주처사대서,시망막포편관찰각조RGC존활정황;매조각개시간점분별처사6지SD대서.채용방차분석대결과진행비교.결과 TUNEL검측현시,술후5 d화2주,각조균가견TUNEL양성세포,기중EPO-PLGA조화EPO조TUNEL양성세포현저감소,기세포조망솔명현소우PLGA조、PBS조급미치료조.술후4주,시망막포편RGC계수현시,정상SD대서RGC밀도위(2387.7±164.9)개/mm2,미치료조위(748.3±58.8)개/mm2,EPO-PLGA조위(1296.7±157.6)개/mm2,EPO조위(1418.5±154.9)개/mm2,PLGA조위(821.7±52.1)개/mm2,PBS조위(804.4±86.4)개/mm2;가견EPO-PLGA조화EPO조교미치료조세포밀도현저증고,구유명현적RGC보호작용(P균＜0.01),이EPO-PLGA조화EPO조간차이무통계학의의(P=0.065).결론 EPO-PLGA완석미구여EPO구유등효적RGC보호작용,저위진일보관찰EPO-PLGA완석미구적장효신경보호작용전정료기출.
Objective To investigate the protective effect of erythropoietin (EPO) encapsulated in poly (L-lactic-co-glycolic acid) (PLGA) microspheres on damaged retinal ganglion cell (RGC) by intravitreal injection after optic nerve crush. Methods Adult SD rats were selected to establish an optic nerve crush model. Immediately after the crush, the animals received intravitreal doses of 10 IU EPO of EPO-PLGA microspheres (EPO-PLGA group), 10 IU EPO (EPO group), blank PLGA microshperes (PLGA group), and PBS (PBS group) or untreated (untreated group). Five days and 2 weeks after crush, apoptotic RGC were detected with TUNEL labeling in frozen retinal sections.Twenty-three days after optic nerve crush, retinal ganglion cells were retrogradely labeled by injecting DiI into the superior colliculi of the brain. The animals were scarificed 4 weeks after the crush. Mounted retinal photographs were assessed for the number of surviving RGC. Six rats were prepared from each group at every time point. Results TUNEL-positive cells could be identified in all groups 5 days and 2 weeks after the operation. However, there were fewer in both the EPO-PLGA and EPO groups. Retinal mounts revealed that the density of RGC was (2387.7±164.9)cells/mm2 in normal adult rats, and (748.3±58.8)cells/mm2 in the untreated group, (1296.7±157.6)cells/mm2 in the EPO-PLGA group, (1418.5±154.9)cells/mm2 in the EPO group, (821.7±52.1)cells/mm2 in the PLGA group, and (804.4±86.4)cells/mm2 in the PBS group 4 weeks after the operation. Compared to the untreated group, the EPO-PLGA and EPO groups showed a significant neuroprotective effect on RGC (P＜0.01), and no significant differences were found between these two groups (P=0.065).Conclusion The protective effect of EPO-PLGA microspheres on RGC is equal to EPO, which provides evidence for further study on the long-term neuroprotective effect of EPO-PLGA microspheres.