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[摘要]
目的:观察先天性白内障小鼠发育过程中视网膜
Acin1基因(apoptotic chromatin condensation inducer 1,凋亡染色体凝聚诱导1)表达的变化及差异,探讨先天性白内障小鼠与正常小鼠视网膜细胞凋亡的差异及晶状体与视网膜发育的联系。
方法:选取先天性白内障小鼠和正常C57BL/6小鼠各15只,其中雌性各10只,雄性各5只,两种小鼠各随机按1雄2雌合笼正常饲养,雌鼠受孕后单独饲养,待其分娩后将对应幼鼠分为先天性白内障组和正常对照组,各组于1,5,9,14,17,21,26,60d分别处理5只幼鼠,取左眼球4%中性甲醛固定行免疫组化检测ACIN1蛋白(ACINUS)表达,右眼取视网膜行PCR检测Acin1 mRNA表达。
结果:Acin1在两组幼鼠视网膜各日龄均有持续表达; 随着视网膜各层细胞的分化,ACIN1蛋白从神经节细胞层、内核层至外核层逐渐表达,以神经节细胞和内核层为主,神经母细胞层未见阳性表达,正常对照组P1~P14d ACIN1阳性细胞逐渐增多,P17d开始减少,P21d之后各层阳性细胞明显减少; 先天性白内障组整体变化趋势同正常对照组类似,P1~P14d阳性细胞数低于正常对照组,P17~P21d阳性细胞数持平且高于正常对照组; 同日龄两组比较,除P17,P26,P60d外,其余差异均有统计学意义(P<0.05); 先天性白内障组内不同日龄总体比较差异有统计学意义(F先白=295.07,P<0.01),两两比较除P1与P5d,P17与P21d外其余各日龄间均有统计学意义(P<0.05); 正常组内不同日龄总体比较差异有统计学意义(F正常=214.21,P<0.01),除P1与P5d外其余各日龄间两两比较均有统计学意义(P<0.05); 先天性白内障组P17d表达量低于正常组P14d差异有统计学意义(P<0.05)。两组Acin1 mRNA变化趋势与蛋白相似,两组同日龄比较,除P17,P21,P60d外差异均有统计学意义(P<0.05); 两组组内不同日龄总体比较差异均有统计学意义(F先白=522.29,P<0.01; F正常=472.05,P<0.01); 两两比较先天性白内障组除P21与P26d外其余各日龄间均有统计学差异(P<0.05),正常组各日龄间两两比较均有统计学差异(P<0.05)。
结论:小鼠视网膜发育过程中存在Acin1时间和空间的差异性表达,先天性白内障晶状体发育障碍可能会影响视网膜细胞Acin1的表达及视网膜细胞的凋亡及发育。
[Key word]
[Abstract]
AIM: To observe the expression of
Acin1(apoptotic chromatin condensation inducer 1)in congenital cataract mouse retina during development and investigate the differences of retinal apoptosis and the connection of lens and retina development between congenital cataract mouse and normal mouse.
METHODS: There were congenital cataract mice(10 female and 5 male)and normal C57BL/6 mice(10 female and 5 male). One male and two female mice were fed in the same cage randomly. The young mice were divided into two groups: congenital cataract group and normal control group. Five young mice were treated each group on 1, 5, 9, 14, 17, 21, 26, 60d. The left eyes were fixed with 4% neutral formalin to detect ACIN1 protein by immunohistochemistry and retinas from right eyes were used to detect the mRNA expression of Acin1.
RESULTS: Acin1 had sustained expression in each group. ACIN1 protein gradually expressed from the ganglion cell layer, inner nuclear layer to the outer nuclear layer following retinal development. It mainly expressed on ganglion cell layer and inner nuclear layer, but not neuroblastoma layer. ACIN1 protein positive cells on P1~P14d increased in normal control group, P17d reduced, after P21d positive cells of each layers decreased. The overall trend was similar in congenital cataract group with normal control group, P1~P14d positive cells count was lower than normal control group, P17-P21d positive cells were flat and higher than the normal control group. Compared with the same day of the two groups, the differences except for P17, P26, P60d were significant(P<0.05). The overall difference was statistically significant in congenital cataract group(Fcataract=295.07, P<0.01); in addition to P1 and P5, P17 and P21, the differences were statistically significant(P<0.05)compared with each other in congenital cataract group. The overall difference was statistically significant in control group(Fnormal=214.21, P<0.01); in addition to P1 and P5d, the difference was statistically significant(P<0.05)compared with each other in control group. The expression of P17d in congenital cataract group was lower compared with that of P14d in control group, the difference was statistically significant(P<0.05). Acin1 mRNA trends of two groups were similar with ACIN1 protein. Compared with the same day of the two groups, the difference was significant except for P17, P21, P60d(P<0.05). The overall difference was statistically significant in each other of the two groups(Fcataract=522.29, P<0.01; Fnormal=472.05, P<0.01). The difference was statistically significant compared with each day in control group(P<0.05). Compared with all the rest of days except for P21 and P26d, the difference was statistically significant in congenital cataract group(P<0.05).
CONCLUSION: Acin1 exist differential expression of time and space in mouse retina during development, congenital cataract crystal developmental disorder may affect the expression of Acin1 and retinal cell apoptosis and development.
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