CAJ | 학술논문

目的观察脑源性神经营养因子(brain-derived nurotrophic factor, BDNF)基因修饰的神经前体细胞(neural progenitor cells, NPCs)玻璃体腔或视网膜下腔移植对视神经轴浆流部分中断视神经病变的治疗作用.方法视网膜神经节细胞经外侧膝状行逆行标记,7天后用40 g力量大小的轴浆流运输钳压迫视神经,制作视神经轴浆流运输部分中断动物模型.动物随机分成3组,将视神经轴浆流运输部分中断的大鼠玻璃体腔内或视网膜下腔注入BDNF基因修饰的NPCs (BDNF-NPCs),注射生理盐水为阴性对照,而注射单纯的NPCs为载体细胞对照.7,15及30天后通过RGCs的平均密度计数以及RGCs的平均凋亡密度计数并进行统计分析比较不同处理组间的差别,对BDNF-NPCs表达的BDNF及其保护作用评价.结果部分轴浆流运输中断后的大鼠玻璃体腔内注射生理盐水的阴性对照及注入单纯NPCs组,7天,15天及30天RGCs的平均密度分别为1885±68,1562±20,1380±7和1837±46,1561±58,1370±16.而在玻璃体腔内注射DNF-NPCs组为2101±15,1809±19和1625±34.在视网膜下腔注入组也获得了相似的结果.对RGCs的平均凋亡密度进行比较,BDNF-NPCs处理组较其它组低.在相应的时间点,BDNF-NPCs处理组较其它组经非参数检验,差异具有统计学意义.结论 BDNF-NPCs注入玻璃体腔或视网膜下腔,都能够表达BDNF并对轴浆流运输部分中断所致的视神经病变起到了明确的保护作用.
목적관찰뇌원성신경영양인자(brain-derived nurotrophic factor, BDNF)기인수식적신경전체세포(neural progenitor cells, NPCs)파리체강혹시망막하강이식대시신경축장류부분중단시신경병변적치료작용.방법시망막신경절세포경외측슬상행역행표기,7천후용40 g역량대소적축장류운수겸압박시신경,제작시신경축장류운수부분중단동물모형.동물수궤분성3조,장시신경축장류운수부분중단적대서파리체강내혹시망막하강주입BDNF기인수식적NPCs (BDNF-NPCs),주사생리염수위음성대조,이주사단순적NPCs위재체세포대조.7,15급30천후통과RGCs적평균밀도계수이급RGCs적평균조망밀도계수병진행통계분석비교불동처리조간적차별,대BDNF-NPCs표체적BDNF급기보호작용평개.결과부분축장류운수중단후적대서파리체강내주사생리염수적음성대조급주입단순NPCs조,7천,15천급30천RGCs적평균밀도분별위1885±68,1562±20,1380±7화1837±46,1561±58,1370±16.이재파리체강내주사DNF-NPCs조위2101±15,1809±19화1625±34.재시망막하강주입조야획득료상사적결과.대RGCs적평균조망밀도진행비교,BDNF-NPCs처리조교기타조저.재상응적시간점,BDNF-NPCs처리조교기타조경비삼수검험,차이구유통계학의의.결론 BDNF-NPCs주입파리체강혹시망막하강,도능구표체BDNF병대축장류운수부분중단소치적시신경병변기도료명학적보호작용.
Objective To investigate in vivo survival of retinal ganglion cells (RGCs) after partial blockage of optic nerve (ON) axoplasmic flow by sub-retinal space or vitreous cavity injection of brain-derived neural factor (BDNF) produced by genetically modified neural progenitor cells (NPCs).Methods Adult Sprague-Dawley (SD) rat RGCs were labeled with granular blue (GB) applied to their main targets in the brain. Seven days later, the left ON was intra-obitally crushed with a 40 g power forceps to partially block ON axoplasmic flow. Animals were randomized to three groups. The left eye of each rat received a sham injection, NPCs injection or an injection of genetically modified neural progenitors producing BDNF (BDNF-NPCs). Seven, 15 and 30 days after ON crush, retinas were examined under a fluorescence microscope. By calculating and comparing the average RGCs densities and RGC apoptosis density, RGC survival was estimated and the neuro-protective effect of transplanted cells was evaluated.Results Seven, 15 and 30 days after crush, in the intra-vitreous injection group, mean RGC densities had decreased to 1885±68, 1562±20, 1380±7 and 1837±46, 1561±58, 1370±16, respectively with sham injection or neural progenitors injection. However, RGCs density in the groups treated with intra-vitreous injection of BDNF-NPC was 2101±15, 1809±19 and 1625±34. Similar results were found in groups after sub-retinal injection. Higher densities were observed in groups treated with BDNF-NPCs. There were statistically significant differences among groups through nonparametric tests followed by the Mann-Whitely test. RGC apoptosis density in BDNF-NPC at each follow-up time was less than in other groups. Conclusions A continuous supply of neurotrophic factors by the injection of genetically modified neural progenitors presents a highly effective approach to counteract optic neuropathy and RGC degeneration after partial ON axoplasmic flow blockage.