PDF(Pubmed)
Abstract：
The retinal mitochondrial injury model in rat has been developed using the mitochondrial oxidative phosphorylation uncoupler, carbonylcyanide m-chlorophenyl hydrazine (CCCP). However, the CCCP-induced murine retinal mitochondrial injury model has not been reported. Here, the optimized conditions for the murine retinal mitochondrial injury model were established by intravitreal injection of different doses of CCCP (0, 2.5, 5, 7.5, 10, 12.5, 15 μg). Indeed, it has been reported that CCCP induces Opa1 cleavage and phosphorylation of ERK in cultured cells and rat retinas. Thus, we measured phosphorylated (p) -Erk and L/S-Opa1 following CCCP-induced retinal injury. Meanwhile, KW6002 (A2A receptor antagonist) pretreatment inhibited retinal injury induced by CCCP at 10 and 15 μg doses differently. Intravitreal injection of 10 μg doses of CCCP can induce apoptosis of retinal ganglion cells and decrease of retinal thickness, but intravitreal injection of 15 μg doses of CCCP is the appropriate dose to study the protective effect of A2A receptor. (1) Dose dependent effects of intravitreal injection of CCCP on the levels of L/S-Opa1 and p-Erk; (2) A2A receptor antagonist (KW6002) only inhibited the apoptosis of ganglion cells, but did not affect the thickness of retina with 10µg dosage of CCCP intravitreal injection; (3) A2A receptor antagonist (KW6002) inhibited the apoptosis of ganglion cells and increased the thickness of retina with 15µg dosage of CCCP intravitreal injection.
摘要：
利用线粒体氧化磷酸化解偶联剂建立了大鼠视网膜线粒体损伤模型,羰基氰化物间氯苯基肼(CCCP)。然而,CCCP诱导的小鼠视网膜线粒体损伤模型尚未见报道。这里,通过玻璃体内注射不同剂量的CCCP（0、2.5、5、7.5、10、12.5、15μg），建立小鼠视网膜线粒体损伤模型的优化条件。的确,据报道，CCCP诱导培养细胞和大鼠视网膜中的Opa1裂解和ERK磷酸化。因此，我们测量了CCCP诱导的视网膜损伤后磷酸化（p）-Erk和L/S-Opa1。同时,KW6002(A2A受体拮抗剂)预处理在10和15μg剂量下不同地抑制CCCP诱导的视网膜损伤。玻璃体内注射10μg剂量的CCCP可诱导视网膜神经节细胞凋亡和视网膜厚度降低，但是玻璃体内注射15μg剂量的CCCP是研究A2A受体保护作用的合适剂量。(1)玻璃体腔注射CCCP对L/S-Opa1和p-Erk水平的剂量依赖性影响;(2)A2A受体拮抗剂(KW6002)仅抑制神经节细胞凋亡,但10µg剂量的CCCP玻璃体内注射不影响视网膜厚度；（3）15µg剂量的CCCP玻璃体内注射A2A受体拮抗剂（KW6002）抑制神经节细胞凋亡并增加视网膜厚度。
