| Objective:In this research,the protective effect of Eerdun-wurile was observed on the retinal ganglion cell in rats with retinal ischemia and reperfusion injury,will expounds the possibility of Eerdun-wurile retinal Ischemia reperfusion injury of disease prevention and cure function.In this study of the effect of Mongolian medicine Eerdun-wurile,to express the Bcl-2,Bax,Fas,Fasl and P53 protein in retina of rats with ischemia-reperfusion injury,and to explore the protective effect of Eerdun-wurile on retinal ischemia-reperfusion injury and its related mechanisms.Methods: Retinal ischemia-reperfusion injury model was induced by elevating intraocular pressure.The effects of drugs in different time and doses on the histopathology of retina will be observed.Several different methods such as Western Blot assay,immunohistochemistry,Fas/Fasl protein assay and computer image analysis system to show the determination of Bcl-2,Bax,Fas,Fasl and P53 protein in rat retina.Results: 1.The changes in Histopathology : After retinal ischemia-reperfusion,edema gradually aggravated,the number of ganglion cells and inner nuclear layer cells decreased,the density of inner and outer nuclear layer decreased,and the arrangement was disordered.After 48 hours and 72 hours,the retina became thinner,the edema was reduced,the number of ganglion cells was scarce,and the nuclear layer was thinner.2.The expression of related factors:Western Blot assay showed that Bcl-2 and Bax and P53 protein were detected in the retina of normal rats.The expression of Bcl-2,Bax and P53 in ischemia-reperfusion group was higher than that in normal group(P < 0.05).The expressions of Bcl-2protein after ischemia-reperfusion were compared in two groups at the same time,it point out that the model group was higher than the normal group,the low-dose group was higher than the model group,the middle-dose group was higher than the low-dose group,and the high-dose group was higher than the middle-dose group(P < 0.01).However,the expression of Bax and P53 at the same time point were compared in two groups,shows the high dose group was higher than the normal group,the middle dose group was higher than the high dose group,the low dose group was higher than the middle dose group,and the model group was higher than the low dose group.The results of Immunohistochemical method showed that there was no Bcl-2,Bax,P53 positive expression in retina 1 hour after ischemia-reperfusion.A small amount of positive expression was found after 6h,but increased at 12 h,peaked at 24 h,and decreased gradually from 48 h to 72 h.The results of Fas/Fasl protein detection showed that Fas was almost not expressed in normal retina tissues.It began to express at 1hour after ischemia-reperfusion,a little at 6 hours,increased at 12 hours,peaked at 24 hours,decreased at 48 hours,and continued to express at 72 hours after ischemia-reperfusion.Fasl expression in normal retina was low,beginning at 1 hour after ischemia-reperfusion,a small amount at 6 hours,increasing at 12 hours,peaking at 24 hours,declining at 48 hours,and still expressing at 72 hours after ischemia-reperfusion.The expression of Bcl-2,Bax,Fas,Fasl and P53 in the retina of rats in Eerdun-wurile group was similar to that in ischemia-reperfusion injury group.However,the expression of Bcl-2 protein was higher in ischemia-reperfusion injury group than in ischemia-reperfusion injury group.The expression of Bax and P53 protein was lower in ischemia-reperfusion injury group than in ischemia-reperfusion injury group.The expression of Fas in 6h,12 h,24 h and 48 h groups was significantly lower than that in ischemia reperfusion injury group(P < 0.01).The expression of Fasl in 12 h,24h and 48 h group was significantly lower than that in the ischemia-reperfusion injury group(P < 0.01).Among these,the expression of each observation index was the highest in the low dose group,followed by the middle dose group,and the lowest in the high dose group.Conclusion: 1.The expression of Bax,Fas,Fasl and P53 protein in retina increased after retinal ischemia-reperfusion injury.Eerdun-wurile will inhibit the increase of retinal ischemia-reperfusion injury and the apoptosis of optic nerve cells,which can be used for the treatment of retinal ischemia-reperfusion injury in clinic.2.Eerdun-wurile can increase the expression of Bcl-2 protein and inhibit the apoptosis of optic nerve cells,which can be used for the treatment of retinal ischemia-reperfusion injury in clinic.3.Eerdun-wurile has protective effect on retinal ischemia-reperfusion injury and can be used in clinical treatment of retinal ischemia-reperfusion injury.4.The high dose group of Eerdun-wurile(0.6g.kg-1)had the best inhibitory effect on optic nerve cell apoptosis,which provided dosage for the study of RIRI mechanism. |
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