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Norbert Kociok, Claudia Brockmann, Elzbieta Jaroslawska-Setlak, Sabrina Dege, Caitlin Corkhill, Antonia M Joussen; Systemic treatment by the Rho-kinase inhibitor Fasudil does not alter the mRNA levels of inflammatory markers after oxygen-induced retinal neovascularization in mice. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3457. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
Rho kinase (ROCK) regulates a variety of cellular events, including endothelial migration and proliferation. The selective ROCK inhibitor Fasudil (C14H17N3O2S HCl),supports neuroprotection in vitro by decreasing the level of the pro-inflammatory factors nitric oxide, IL-1β, IL-6, and TNFa in cultured microglia. We investigated the effects of Fasudil on the mRNA expression of these factors in an animal model of oxygen-induced retinopathy.
In a model of oxygen-induced retinal neovascularization mice received daily intraperitoneal injections of Fasudil at a dose of 10mg/kg from P12 to P16. The mRNA levels of TNFa, IL-1b, IL-6, Nos-1/nNos, Nos-2/iNos, Nos-3/eNos, VEGF-A were determined by qPCR. Retinal vasculature on flatmounts was analyzed after staining with isolectin IB4.
The mRNA level of IL-1b, IL-6, and TNFa are elevated at P14 in O2-treated mice. The level decreased at P17 for IL-1b and IL-6 to the normoxic level, but increased further for TNFa, whereas at P21 normoxic mRNA level was reached for all three genes. Relative hypoxia led to a downregulation of Nos-1/nNos mRNA at P14 and P17, which at P21 reaches normoxic level again. Irrespective to the treatment there is an equal high expression level for Nos-2/iNos mRNA at P14, which decreases to a lower level in O2-treated mice. At P21 the Nos-2/iNos mRNA expression is very similar in O2-treated and untreated mice. For Nos-3/eNos there is no treatment-related difference in the mRNA expression as the mRNA level is decreasing from P14 to P21. Injecting Fasudil did not alter the mRNA level of the analyzed genes at P14 or P17.At P14, the retinas of the control group showed a mean (±SD) avascular area of 29.6 ± 6.8 % of total retinal area (NaCl injected mice: 24.5 ± 4.7 %; Fasudil injected mice: 25.6 ± 2.3 %). No significant differences were found between the groups (p = 0.331, p = 0.682, p = 0.562, respectively).
Our findings demonstrate that a systemic application of Fasudil did not alter the mRNA levels of inflammatory markers in mice with oxygen-induced retinopathy, and did not decrease retinal neovascularization; but that for this purpose intravitreal injection may be necessary. The results of our study makes it clear that the effects of ROCK inhibition are complex and further studies are necessary to analyze its potential therapeutic effects.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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