腺苷 A 受体拮抗剂在高氧期发挥作用，对视网膜病变起到保护作用。
Adenosine A receptor antagonists act at the hyperoxic phase to confer protection against retinopathy.
机构信息
Institute of Molecular Medicine, School of Optometry and Ophthalmology and Eye Hospital, Wenzhou Medical University, 270 Xueyuan Road, Wenzhou, 325027, Zhejiang, China.
State Key Laboratory Cultivation Base and Key Laboratory of Vision Science, Ministry of Health, China and Zhejiang Provincial Key Laboratory of Ophthalmology and Optometry, Wenzhou, Zhejiang, China.
出版信息
Mol Med. 2018 Jul 31;24(1):41. doi: 10.1186/s10020-018-0038-1.
BACKGROUND
Retinopathy of prematurity (ROP) remains a major cause of childhood blindness and current laser photocoagulation and anti-VEGF antibody treatments are associated with reduced peripheral vision and possible delayed development of retinal vasculatures and neurons. In this study, we advanced the translational potential of adenosine A receptor (AR) antagonists as a novel therapeutic strategy for selectively controlling pathological retinal neovascularization in oxygen-induced retinopathy (OIR) model of ROP.
METHODS
Developing C57BL/6 mice were exposed to 75% oxygen from postnatal (P) day 7 to P12 and to room air from P12 to P17 and treated with KW6002 or vehicle at different postnatal developmental stages. Retinal vascularization was examined by whole-mount fluorescence and cross-sectional hematoxylin-eosin staining. Cellular proliferation, astrocyte and microglial activation, and tip cell function were investigated by isolectin staining and immunohistochemistry. Apoptosis was analyzed by TUNEL assay. The effects of oxygen exposure and KW6002 treatment were analyzed by two-way ANOVA or Kruskal-Wallis test or independent Student's t-test or Mann-Whitney U test.
RESULTS
The AR antagonist KW6002 (P7-P17) did not affect normal postnatal development of retinal vasculature, but selectively reduced avascular areas and neovascularization, with the reduced cellular apoptosis and proliferation, and enhanced astrocyte and tip cell functions in OIR. Importantly, contrary to our prediction that AR antagonists were most effective at the hypoxic phase with aberrantly increased adenosine-AR signaling, we discovered that the AR antagonist KW6002 mainly acted at the hyperoxic phase to confer protection against OIR as KW6002 treatment at P7-P12 (but not P12-P17) conferred protection against OIR; this protection was observed as early as P9 with reduced avascular areas and reduced cellular apoptosis and reversal of eNOS mRNA down-regulation in retina of OIR.
CONCLUSIONS
As ROP being a biphasic disease, our identification of the hyperoxic phase as the effective window, together with selective and robust protection against pathological (but not physiological) angiogenesis, elevates AR antagonists as a novel therapeutic strategy for ROP treatment.
背景
早产儿视网膜病变（ROP）仍然是儿童失明的主要原因，目前的激光光凝和抗 VEGF 抗体治疗与周边视力下降以及视网膜血管和神经元发育延迟有关。在这项研究中，我们推进了腺嘌呤 A 受体（AR）拮抗剂作为一种新的治疗策略的转化潜力，用于选择性控制氧诱导的 ROP 模型中的病理性视网膜新生血管形成。
方法
对 C57BL/6 幼鼠进行实验，使其在出生后（P）第 7 天至 P12 天暴露于 75%氧气中，然后在 P12 天至 P17 天暴露于室内空气中，并在不同的出生后发育阶段用 KW6002 或载体处理。通过全视网膜荧光和横切苏木精-伊红染色检查视网膜血管生成。通过异硫氰酸荧光素染色和免疫组织化学研究细胞增殖、星形胶质细胞和小胶质细胞激活以及尖端细胞功能。通过 TUNEL 分析检测细胞凋亡。通过双因素方差分析或 Kruskal-Wallis 检验或独立学生 t 检验或曼-惠特尼 U 检验分析氧暴露和 KW6002 治疗的效果。
结果
AR 拮抗剂 KW6002（P7-P17）不影响视网膜血管的正常出生后发育，但选择性地减少无血管区和新生血管形成，减少细胞凋亡和增殖，并增强 OIR 中的星形胶质细胞和尖端细胞功能。重要的是，与我们的预测相反，即 AR 拮抗剂在缺氧阶段具有异常增加的腺苷-AR 信号时最有效，我们发现 AR 拮抗剂 KW6002 主要在高氧阶段发挥作用，以防止 OIR，因为 KW6002 治疗在 P7-P12（而不是 P12-P17）时可防止 OIR；这种保护作用早在 P9 时就观察到，无血管区减少，细胞凋亡减少，视网膜中 eNOS mRNA 下调逆转。
结论
由于 ROP 是一种双相疾病，我们确定高氧期为有效窗口期，以及对病理性（而非生理性）血管生成的选择性和强大保护作用，使 AR 拮抗剂成为 ROP 治疗的一种新的治疗策略。
Zotero 插件