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Abstract #27362 Published in IGR 12-4
Expression of hypoxia-inducible factor-1(alpha), inducible nitric oxide synthase and cyclooxygenase-2 in rat retina with chronic high ocular pressure
Gui D-M; Yang Y; Li X; Gao D-WChinese Ophthalmic Research 2010; 28: 805-809
Background: Researches showed that hypoxia-inducible factor-1(alpha) (HIF-1(alpha)) is a transcription factor with the close relationship to hypoxia. So to study the role of HIF-1 and its relevant target gene in the damage of glaucomatous retinal ganglion cells is very important. Objective: The aim of this study was to investigate the expressions of downstream gene of HIF-1, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in normal retina and ocular hypertension retina, and investigate the relation of glaucomatous retinal degeneration and hypoxia-responsive gene regulation. Methods: Chronic ocular hypertension models were created in both eyes of 50 SPF Wistar rats by cautering two episcleral veins unilaterally with electric coagulator. Only bulbar conjunctiva was cut in other 20 eyes of 10 rabbits as sham operative group. Intraocular pressure (IOP) of animal models was measured with TONO-PEN II tonometor. The animals were sacrificed respectively in 3,7,14,21 and 28 days after establishment of models. Expressions of HIF-1(alpha), iNOS and COX-2 protein in retina were detected by Western blot, and the expressions of HIF-1(alpha), iNOS and COX-2 mRNA were assessed by reverse transcriptase-polymerase chain reaction (RT-PCR). Use of animals followed the Regulation for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. Results: The IOP was more 40% after cauterization of episclera in model eyes. RT-PCR revealed that HIF-1(alpha), iNOS and COX-2 mRNA were faintly expressed in rats retinas of sham operative group. The expressive levels of HIF-1(alpha), iNOS and COX-2 mRNA were enhanced in 3,7,14,21 and 28 days after operation with statistically significant differences in comparison with sham operative group (P < 0.05, P < 0.01). Western blot assay indicated that the expressions of HIF-1(alpha), iNOS and COX-2 proteins showed the same pattern to the expressions of HIF-1(alpha), iNOS and COX-2 mRNA. The peak expressions of HIF-1(alpha), iNOS and COX-2 genes were on 7 days after IOP elevated then declined but still kept a higher level until 28 days. Conclusion: HIF-1(alpha), iNOS and COX-2 participates in signal transduction of retinal neurons during retinal injury on chronic high ocular pressure.
D.-M. Gui. Department of Ophthalmology, Affiliated Shengjing Hospital, China Medical University, Shenyang 110003, China. guidongmei2004@yahoo.com.cn
Classification:
5.1 Rodent (Part of: 5 Experimental glaucoma; animal models)
3.6 Cellular biology (Part of: 3 Laboratory methods)
3.5 Molecular biology incl. SiRNA (Part of: 3 Laboratory methods)
11.8 Neuroprotection (Part of: 11 Medical treatment)
