UNASSIGNED: AOH rat model was performed by anterior chamber perfusion and either received an intravitreal injection with β-aminopropionitrile (BAPN) or normal saline. After 2wk, Quantification of survival RGCs in the retina was performed using Retrograde FluoroGold labeling. The mRNA expression levels of LOX, LOXL1-4, collagen 1a1 (Col1a1), collagen 3a1 (Col3a1), collagen4a1 (Col4a1), elastin (Eln), fibronectin1 (Fbn1), fibronectin4 (Fbn4) were determined by RT-qPCR. LOX expression was determined by Western blot (WB) analysis and immunohistochemistry. The RNA expression of LOX, Eln and Col1a1 in RGCs retrograde-labeled with 1,1\'-dioctadecyl-3,3,3\',3\' tetra-methylindocarbocyanine perchlorate(DiI)that selected through FACS sorting were determined by RT-qPCR analysis. Changes of the retinal function were detected by Electroretinogram (ERG) analysis.
UNASSIGNED: Results showed that significant LOX overexpression and loss of RGCs related to IOP exposure in AOH retinas. PCR analysis indicated significant increased mRNA level of Col1a1, Col3al and Eln in AOH retinas. Significant increase mRNA expression of LOX, Col1a1 and Eln in the RGCs were observed in AOH group compared with CON group. AOH rats injected with BAPN showed a significant decrease in LOX expression, reduced the loss of RGCs and retinal function damage.
UNASSIGNED: The results demonstrated that changes of LOX and specific ECM components in retina were correlated with AOH. Findings from this study indicated that preventing LOX over-expression may be protective against RGCs loss and retinal function damage in AOH animal model.
■AOH大鼠模型通过前房灌注进行,并接受玻璃体内注射β-氨基丙腈(BAPN)或生理盐水。2wk之后,使用逆行荧光金标记进行视网膜中存活RGC的定量。LOX的mRNA表达水平,LOXL1-4,胶原蛋白1a1(Col1a1),胶原蛋白3a1(Col3a1),胶原4a1(Col4a1),弹性蛋白(Eln),纤连蛋白1(Fbn1),通过RT-qPCR测定纤连蛋白4(Fbn4)。通过蛋白质印迹(WB)分析和免疫组织化学测定LOX表达。LOX的RNA表达,Eln和Col1a1在用1,1'-双十八烷基-3,3'逆行标记的RGC中,通过RT-qPCR分析确定通过FACS分选选择的3''四甲基吲哚草青高氯酸盐(DiI)。通过视网膜电图(ERG)分析检测视网膜功能的变化。
■结果表明,在AOH视网膜中,LOX的过表达和RGC的丢失与IOP暴露有关。PCR分析表明,AOH视网膜中Col1a1,Col3al和Eln的mRNA水平显着增加。显著增加LOX的mRNA表达,与CON组相比,AOH组RGC中观察到Col1a1和Eln。注射BAPN的AOH大鼠LOX表达显著降低,减少RGCs的丢失和视网膜功能损伤。
■结果表明,视网膜中LOX和特定ECM成分的变化与AOH相关。这项研究的结果表明,在AOH动物模型中,防止LOX过表达可能对RGC损失和视网膜功能损伤具有保护作用。