Effects of a Rho kinase inhibitor on the sequential expression of ICAM-1, HIF-1α, Bcl-2 and caspase-3 in the retina of rats with oxygen-induced retinopathy

Wang,Haiying; Chen,Xiaolong

doi:10.3892/ijmm.2013.1410

Effects of a Rho kinase inhibitor on the sequential expression of ICAM-1, HIF-1α, Bcl-2 and caspase-3 in the retina of rats with oxygen-induced retinopathy

Authors:
- Haiying Wang
- Xiaolong Chen
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Affiliations: Department of Ophthalmology, The Fourth People's Hospital of Shenyang, Shenyang, Liaoning 110031, P.R. China, Department of Ophthalmology, Shengjing Hospital Affiliated to China Medical University, Shenyang, Liaoning 110004, P.R. China
Published online on: June 5, 2013 https://doi.org/10.3892/ijmm.2013.1410
Pages: 457-463

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Abstract

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.

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1	Pastor JC, de la Rúa ER and Martín F: Proliferative vitreoretinopathy: risk factors and pathobiology. Prog Retin Eye Res. 21:127–144. 2002. View Article : Google Scholar : PubMed/NCBI
2	Hartnett ME: The effects of oxygen stresses on the development of features of severe retinopathy of prematurity: knowledge from the 50/10 OIR model. Doc Ophthalmol. 120:25–39. 2010. View Article : Google Scholar : PubMed/NCBI
3	Penn JS: Retinal and Choroidal Angiogenesis. Springer; Dordrecht: pp. 1192008
4	Näpänkangas U, Lindqvist N, Lindholm D and Hallböök F: Rat retinal ganglion cells upregulate the pro-apoptotic BH3-only protein Bim after optic nerve transection. Brain Res Mol Brain Res. 120:30–37. 2003.PubMed/NCBI
5	Zhang Z, Qin X, Zhao X, Tong N, Gong Y, Zhang W and Wu X: Valproic acid regulates antioxidant enzymes and prevents ischemia/reperfusion injury in the rat retina. Curr Eye Res. 37:429–437. 2012. View Article : Google Scholar : PubMed/NCBI
6	Cellerino A, Bähr M and Isenmann S: Apoptosis in the developing visual system. Cell Tissue Res. 301:53–69. 2000. View Article : Google Scholar
7	Ozaki H, Yu AY, Della N, Ozaki K, Luna JD, Yamada H, Hackett SF, Okamoto N, Zack DJ, Semenza GL and Campochiaro PA: Hypoxia inducible factor-1alpha is increased in ischemic retina: temporal and spatial correlation with VEGF expression. Invest Ophthalmol Vis Sci. 40:182–189. 1999.PubMed/NCBI
8	Joussen AM, Murata T, Tsujikawa A, Kirchhof B, Bursell SE and Adamis AP: Leukocyte-mediated endothelial cell injury and death in the diabetic retina. Am J Pathol. 158:147–152. 2001. View Article : Google Scholar : PubMed/NCBI
9	Barouch FC, Miyamoto K, Allport JR, Fujita K, Bursell SE, Aiello LP, Luscinskas FW and Adamis AP: Integrin-mediated neutrophil adhesion and retinal leukostasis in diabetes. Invest Ophthalmol Vis Sci. 41:1153–1158. 2000.PubMed/NCBI
10	Rothlein R, Dustin ML, Marlin SD and Springer TA: A human intercellular adhesion molecule (ICAM-1) distinct from LFA-1. J Immunol. 137:1270–1274. 1986.
11	Samarin S and Nusrat A: Regulation of epithelial apical junctional complex by Rho family GTPases. Front Biosci. 14:1129–1142. 2007.PubMed/NCBI
12	Smith A, Bracke M, Leitinger B, Porter JC and Hogg N: LFA-1-induced T cell migration on ICAM-1 involves regulation of MLCK-mediated attachment and ROCK-dependent detachment. J Cell Sci. 116:3123–3133. 2003. View Article : Google Scholar : PubMed/NCBI
13	Leung T, Chen XQ, Manser E and Lim L: The p160 RhoA-binding kinase ROK alpha is a member of a kinase family and is involved in the reorganization of the cytoskeleton. Mol Cell Biol. 16:5313–5327. 1996.PubMed/NCBI
14	Riento K and Ridley AJ: Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol. 4:446–456. 2003. View Article : Google Scholar : PubMed/NCBI
15	Amano M, Nakayama M and Kaibuchi K: Rho-kinase/ROCK: A key regulator of the cytoskeleton and cell polarity. Cytoskeleton (Hoboken). 67:545–554. 2010. View Article : Google Scholar : PubMed/NCBI
16	Warner SJ and Longmore GD: Distinct functions for Rho1 in maintaining adherens junctions and apical tension in remodeling epithelia. J Cell Biol. 185:1111–1125. 2009. View Article : Google Scholar : PubMed/NCBI
17	Brady DC, Alan JK, Madigan JP, Fanning AS and Cox AD: The transforming Rho family GTPase Wrch-1 disrupts epithelial cell tight junctions and epithelial morphogenesis. Mol Cell Biol. 29:1035–1049. 2009. View Article : Google Scholar : PubMed/NCBI
18	Olson MF: Applications for ROCK kinase inhibition. Curr Opin Cell Biol. 20:242–248. 2008. View Article : Google Scholar : PubMed/NCBI
19	Abman SH: Recent advances in the pathogenesis and treatment of persistent pulmonary hypertension of the newborn. Neonatology. 91:283–290. 2007. View Article : Google Scholar : PubMed/NCBI
20	Budzyn K, Marley PD and Sobey CG: Targeting Rho and Rho-kinase in the treatment of cardiovascular disease. Trends Pharmacol Sci. 27:97–104. 2006. View Article : Google Scholar : PubMed/NCBI
21	Shimokawa H: Rho-kinase as a novel therapeutic target in treatment of cardiovascular diseases. J Cardiovasc Pharmacol. 39:319–327. 2002. View Article : Google Scholar : PubMed/NCBI
22	Hata Y, Miura M, Nakao S, et al: Antiangiogenic properties of fasudil, a potent Rho-Kinase inhibitor. Jpn J Ophthalmol. 52:16–23. 2008. View Article : Google Scholar : PubMed/NCBI
23	Bryan BA, Dennstedt E, Mitchell DC, et al: RhoA/ROCK signaling is essential for multiple aspects of VEGF-mediated angiogenesis. FASEB J. 24:3186–3195. 2010. View Article : Google Scholar : PubMed/NCBI
24	Arita R, Hata Y, Nakao S, et al: Rho kinase inhibition by fasudil ameliorates diabetes-induced microvascular damage. Diabetes. 58:215–226. 2009. View Article : Google Scholar : PubMed/NCBI
25	Yin L, Morishige K, Takahashi T, et al: Fasudil inhibits vascular endothelial growth factor-induced angiogenesis in vitro and in vivo. Mol Cancer Ther. 6:1517–1125. 2007. View Article : Google Scholar : PubMed/NCBI
26	Penn JS, Tolman BL and Lowery LA: Variable oxygen exposure causes preretinal neovascularization in the newborn rat. Invest Ophthalmol Vis Sci. 34:576–585. 1993.PubMed/NCBI
27	Kim WT and Suh ES: Retinal protective effects of resveratrol via modulation of nitric oxide synthase on oxygen-induced retinopathy. Korean J Ophthalmol. 24:108–118. 2010. View Article : Google Scholar : PubMed/NCBI
28	Sharp FR, Ran R, Lu A, et al: Hypoxic preconditioning protects against ischemic brain injury. NeuroRx. 1:26–35. 2004. View Article : Google Scholar : PubMed/NCBI