| Objective The aim of the study was focused on the influence of Sodium Ozagrel inretinal ischemia reperfusion injury in rabbit reinal ischemia reperfusion injury (retinalischemia-reperfusion injury RIRI) model with injection of Sodium Ozagrel through earvein, to explore the neuprotective mechanism of Sodium Ozagrel on retinal ischemiareperfusion injury and its effect on Caspase-3and Bcl-2expression, provide a theoreticalbasis for the treatment of retinal ischemic injury in clinical environment.Method A total of78healthy male rabbits from Japanese, weighing2.0~2.5kg, wererandomly divided into three groups: normal group (6rabbits), ischemia-reperfusion group(IR,36rabbits) and Ozagrel therapy group (36rabbits). The last two groups were dividedinto6h,12h,24h,48h,72h and120h groups (6rabbits in each group). Normal groupreceived no treatment; IR group prepared ischemia-reperfusion model only by improvedpressure of intraocular anterior with booster device;(Ozagrel+IR) treatment groupimmediately injection ozagrel sodium after ischemia-reperfusion,25mg/kg,1times/d.normal and injury groups injected the same dose of balanced salt solution. Rabbits ineach group were sacrificed with embolism immediately afte reperfusion for six times byinjection of10ml air to ear vein, then the ischemia-reperfusion eyes wereremoved.Retinal tissue of different time points were stained by HE staining and thenretinal morphological changes were observed. Apoptotic genes Caspase-3and Bcl-2weredetected by immunohistochemical staining and neuronal apoptosis was detected byTUNEL method. Results obtained for image analysis, the data were analyzed bystatistical software SPSS (version17.0).Result1HE staining: In normal group, levels of retinal tissue were clear, the ganglioncells were densely packed with monolayer and cells volume were larger. In the IR group,the numbers of ganglion cells decreased compared with the control group, retinal edemaintensified with time delayed, cells in layers arranged slightly loose, vacuolardegeneration.Reperfusion for72h, retinal edema, the number of ganglion cells reduced.After120h, retinal edema disappeared, but the whole retinal thinning. In Ozagrel therapygroup, the degree of numbers reduction of ganglion cells and cells damage easedcompared with IR group.2Expression changes of Caspase-3and Bcl-2: there were no positive cells in the normal group; after reperfusion for6h the expression increased,positive parts mainly located in the ganglion cell layer; reperfusion for12h the expressionincreased and reached a peak on24h, positive parts mainly located in the ganglion celllayer, inner nuclear layer also had a small amount of expression but no expression in theouter nuclear layer; after reperfusion for48h the expression gradually decreased, positiveparts mainly located in the ganglion cell layer; reperfusion for72h little expression couldbe observed; to reperfusion for120h, there were almost no positive cells in retina. Eachobservation indicators in treatment group were similarly to the injury group. Same timepoints in the treatment group and injury group, the expression Bcl-2was significantlyincreased (except for the120h group) and Caspase-3was significantly decreased(P<0.05). In the injury group and the treatment group between different time pointsnumbers of positive cells were significant differences (P<0.01).3TUNEL assay: in thenormal group, there was no positive expression of apoptotic cells. Reperfusion for6hthere was small amount of positive expression of apoptotic cells and significantlyincreased on12h, which reached a peak on24h, positive pars mainly located in theganglion cell layer and inner nuclear layer. Reperfusion for48h, expression of apoptoticcells began to decrease and significantly reduced on72h; there showed no expression ofapoptotic cells after120h.The positive expression trend of apoptotic cells in the treatedgroup was basically similar with the injury group, but the amount of expressionsignificantly reduced. Comparison between injury and treatment groups in the same timepoint, the numbers of apoptotic cells in each time point of12h,24h,48h and72h weresignificant difference (P<0.01), whereas there was difference in6h,120h group (P<0.05).In the injury group and the treatment group between different time points numbers ofapoptotic cells were significant differences (P<0.01).Conclusion Ozagrel has a significant protective effect to nerve cells of the retina whenretinal ischemia-reperfusion injury occurred. Ozagrel can downregulate the expression ofCaspase-3and increase Bcl-2, thus inhibit apoptosis of retinal neurons when RIRI. |
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