Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning 110031, P.R. China.
Int J Mol Med. 2013 Aug;32(2):457-63. doi: 10.3892/ijmm.2013.1410. Epub 2013 Jun 5.
The aim of the present study was to evaluate the effects of blocking the Rho kinase pathway on non-perfused regions and angiogenesis in the retina of rats using a rat model of oxygen-induced retinopathy (OIR) by observing the sequential expression of intercellular adhesion molecule-1 (ICAM-1), hypoxia-inducible factor-1 (HIF-1), B-cell lymphoma/leukemia-2 gene (Bcl-2) and caspase-3 mRNA following the administration of the Rho kinase inhibitor, fasudil (FSD). A total of 240 newborn rats were randomly divided into a normoxia control (N) group, a hyperoxia (H) group and a H + FSD (HF) group. The rats were sacrificed, and the eyes were enucleated from postnatal day (P)12 to P21. Samples were prepared for retinal flat mounts, mRNA and protein quantification. On P14, a higher number of circuitous retinal veins was observed in the H group compared with the HF group. In the HF group, the avascular area was significantly reduced compared with the H group on P18 (P<0.01). In the HF group, the mRNA expression of Bcl-2 was significantly increased on P15 compared with the N and H group (P<0.01). On P15 and P17 in the H group and on P13 in the HF group, the mRNA expression of ICAM‑1 was significantly increased compared with the other groups (P<0.05). In the H and HF group, the expression of HIF-1α was significantly increased on P12 compared with the N group (P<0.05). On P19 and P21, HIF-1α expression was significantly increased to a maximum level in the HF group compared with the H and N group (P<0.01). In conclusion, these results suggest that FSD inhibits the expression of ICAM-1, assisting in the release of Bcl-2, suppressing caspase-3. In the HF group, the retinal flat mounts revealed that FSD had a vasorelaxant effect. On P18, a double-layered retinal vascular network was formed, and the number of non-perfused regions was significantly reduced. However, the late-phase peak expression of HIF-1α resulted in an inevitable increase in vascular endothelial growth factor expression and further accelerated neovascularization and vascular reconstruction in the immature retinal model.
本研究旨在通过观察大鼠氧诱导视网膜病变（OIR）模型中细胞间黏附分子-1（ICAM-1）、缺氧诱导因子-1（HIF-1）、B 细胞淋巴瘤/白血病-2 基因（Bcl-2）和半胱天冬酶-3 mRNA 的序贯表达，评估 Rho 激酶通路阻断对未灌注区和视网膜血管生成的影响，使用 Rho 激酶抑制剂 fasudil（FSD）。共 240 只新生大鼠随机分为常氧对照组（N）、高氧组（H）和 H+FSD（HF）组。从出生后第 12 天（P）到第 21 天，处死大鼠并取出眼球。制备视网膜平铺标本，进行 mRNA 和蛋白定量。在 P14 时，与 HF 组相比，H 组中出现了更多的迂回视网膜静脉。在 HF 组中，与 H 组相比，P18 时无血管区明显减少（P<0.01）。在 HF 组中，与 N 组和 H 组相比，P15 时 Bcl-2mRNA 表达明显增加（P<0.01）。在 H 组 P15 和 P17 以及 HF 组 P13 时，ICAM-1mRNA 表达与其他组相比明显增加（P<0.05）。在 H 和 HF 组中，与 N 组相比，P12 时 HIF-1α 的表达明显增加（P<0.05）。在 P19 和 P21 时，HF 组 HIF-1α 的表达达到最大值，与 H 组和 N 组相比明显增加（P<0.01）。综上所述，这些结果表明 FSD 抑制了 ICAM-1 的表达，有助于 Bcl-2 的释放，抑制 caspase-3。在 HF 组中，视网膜平铺显示 FSD 具有血管舒张作用。在 P18 时，形成了双层视网膜血管网络，未灌注区的数量明显减少。然而，HIF-1α 的晚期峰值表达导致血管内皮生长因子表达的不可避免增加，并进一步加速了未成熟视网膜模型中的新生血管形成和血管重建。
