The Effects Of Huoxuehuayu Prescription On Retinal Hypoxia-inducible Factor-1 Alpha In Acute Intraocular Hypertension Of Rats

Objective To evaluate the protective role of Huoxuehuayu prescription on rat's retina injury induced by acute intraocular hypertension and to observe the expression changes of HIF-1awith the prescription.Methods 140 Wistar rats were randomly divided into four groups, normal control group including 10 rats, punctured group including 10 rats, pressurized group and treated group including 60 rats respectively. The rats in punctured group were punctured in the left eyes. The acute intraocular hypertension model was created by anterior chamber perfusion in rats'left eyes for 60 minutes in pressurized and treated groups. There were 10 rats were sacrificed at 2h,12h,1d,3d,7d,14d randomly and respectively after being pressurized in pressurized group. The rats in treated group were treated with Huoxuehuayu prescription immediately after pressurizing and were sacrificed at 2h,12h,1d,3d,7d,14d respectively after pouring drugs.5 random rats'retina of every 10 rats were H-E stained and observed under light microscope, and the protein level of HIF-la were detected by immunohistochemistry; HIF-1a mRNA in rats' retina were detected by Real-Time quantitative reverse transcription polymerase chain reaction(Real-Time qRT-PCR) using the rest rats' retina.Results1. The histological changes under light microscope:2h after pressurizing, mild edema appeared in the retina; 12h, the edema became severe and retina became thick; 1d, the retina were dropsical the most severely, and the RGCs were loosen; 3d, the dropsical condition was alleviative, and the ganglion cell layer became thin mildly; 7d, the edema was disappeared generally, the structure and arrangement were disorganized, and the rest number of RGCs was smaller; 14d, the retina became atrophic and thinner, the RGCs were few in number, and the shape of nucleus were irregular. The changing trend in the pressurized and treated groups was similar to that of the pressurized groups, but the extent of injury in the former groups were less severe than the latter groups of the same time stages except for the 2h,12h.2. IHC results:2h of the pressurized groups, there was slight expression of HIF-1a protein in the GCL, and got to the peak at 12h, then decreased gradually. The expressions of HIF-1a in the pressurized and treated groups were less than those in the pressurized groups of the same time stages except for the 2h,12h.3. Real-Time qRT-PCR results:The expression of HIF-1a mRNA in 2h of the pressurized groups was higher than the normal group, got to the peak at 12h and decreased gradually; the expressions in the pressurized and treated groups were less than those in the pressurized groups of the same time stages except for the 2h,12h.Conclusion:1. The expression of HIF-la in the retina of acute intraocular hypertension rats increases and it may play an important role in the pathogenesis of glaucoma.2. The Huoxuehuayu prescription can protect the rat's retina which was injured by acute intraocular hypertension. This protective role may be concerned with that it can reduce the expression of endogenous HIF-la in retina.