CAJ | 학술논문

目的研究大鼠视网膜急性缺血中视网膜节细胞(RGCs)死亡时脑红蛋白(Nsb)的表达.方法实验研究.采用特异性结扎大鼠视网膜动脉的方法造成视网膜急性缺血模型,按照结扎时间不同(0、15、30、60 min)分为A、B、C、D组,每组10只大鼠,均以左眼为实验眼.每组随机选取3只大鼠视网膜用于免疫印迹法检测,4只大鼠视网膜做HE染色节细胞计数及免疫组化染色荧光强度分析,3只大鼠做免疫电子显微镜观察.采用单因素方差分析对不同结扎时间Ngb蛋白定量及RGCs数目进行统计学分析,采用KSD-t检验进行两两比较.结果视网膜完全结扎后,A、B、c、D组Ngb蛋白表达量分别为1.439±0.014、2.023±0.134、1.416±0.030及1.073±0.064;RGCs计数结果分别为4368±124、4296 ±96、4132±104、3973±115,组间比较差异有统计学意义(F=79.548,10.191,P<0.05).B组表达最高,之后逐渐降低,C组接近正常,此时与A组比较RGCs数量开始减少,D组Ngb的表达已明显低于A组,与A组比较RGCs数量进一步减少.在视网膜各层中,Ngb的表达以RGCs层为最高,其次为内丛状层与外丛状层.RGCs中Ngb主要分布在胞质,C组线粒体外室和线粒体嵴也发现有Ngb表达,而内核层及外核层细胞的胞质内始终未见Ngb的表达.结论 Ngb在大鼠RGCs急性缺血死亡时表达快速升高,主要表达于RGCs的胞质内,与RGCs的生存状态关系密切.
목적 연구대서시망막급성결혈중시망막절세포(RGCs)사망시뇌홍단백(Nsb)적표체.방법 실험연구.채용특이성결찰대서시망막동맥적방법조성시망막급성결혈모형,안조결찰시간불동(0、15、30、60 min)분위A、B、C、D조,매조10지대서,균이좌안위실험안.매조수궤선취3지대서시망막용우면역인적법검측,4지대서시망막주HE염색절세포계수급면역조화염색형광강도분석,3지대서주면역전자현미경관찰.채용단인소방차분석대불동결찰시간Ngb단백정량급RGCs수목진행통계학분석,채용KSD-t검험진행량량비교.결과 시망막완전결찰후,A、B、c、D조Ngb단백표체량분별위1.439±0.014、2.023±0.134、1.416±0.030급1.073±0.064;RGCs계수결과분별위4368±124、4296 ±96、4132±104、3973±115,조간비교차이유통계학의의(F=79.548,10.191,P<0.05).B조표체최고,지후축점강저,C조접근정상,차시여A조비교RGCs수량개시감소,D조Ngb적표체이명현저우A조,여A조비교RGCs수량진일보감소.재시망막각층중,Ngb적표체이RGCs층위최고,기차위내총상층여외총상층.RGCs중Ngb주요분포재포질,C조선립체외실화선립체척야발현유Ngb표체,이내핵층급외핵층세포적포질내시종미견Ngb적표체.결론 Ngb재대서RGCs급성결혈사망시표체쾌속승고,주요표체우RGCs적포질내,여RGCs적생존상태관계밀절.
Objective To investigate the intra-retinal expression of neuroglobin (Ngb) and death of retinal ganglion cells (RGCs) in acute retina ischemia rats. Methods It was an experimental study. The acute retina ischemia model was established by specific hypodesised left retina artery of Sprague-Dawdey rats. Forty rats were divided into four groups (0, 15, 30, 60 min) by the time of retina ischemia. Every group has 10 rats, in one group random 3 rats were detected by Western blotting;4 rats were detected by ganglion cell counted by hematoxylin and eosin stain and immunohistochemisty fluorescence intensity analysis. The rest 3 rats were detected by Western blotting. The difference among different data were analyzed statistically by One-factor analysis of variance and LSD-t analysis. Results The intra-retinal expression of Ngb reached maximum after acute ischemia 15 minute (P = 0. 000). then the expression began decreasing. After 30 minute acute ischemia, the expression of Ngb had approached normal (P =0.728), while, the cell number of RGCs began lower than 0 min group (P = 0.011) ; after 60 minute acute ischemia, the expression of Ngb had been obviously lower than 0 min group (P =0. 001), the cell number of RGCs had been further lower than 0 min group (P =0. 000). The expression of Ngb in RGCs layer was highest in rat retina. The expression in inner plexiform layer and external plexiform layer were lower than the former. The expression of Ngb RGCs was mostly intracytoplasm. After 30 minute acute ischemia, the expression of Ngb were detected in mitochondrial outer compartment and mitochondrial cristae, but in cytoplasm of inner nuclear layer and outer nuclear layer the Ngb was not found. Conclusion Ngb quickly steps-up when RGCs die in acute retina ischemia, and mainly expresses intracytoplasm of RGCs. It has tense relationships with nerve cells' survival in hypoxia.