| Objective The increase of intraocular pressure by the animal research,and to establish human retinal ischemia reperfusion injury(RIRI)animal model similar to that of animal erythropoietin(rhEPO)combined with fuming decoction.The changes of retinal structure of RIRI after administration of animal retinal cells Bcl-2,Bax level,and the apoptosis of nerve cells were observed,analysis of the mechanism of action of the drugs on the RIRI,for the clinical diagnosis and treatment of RIRI lay a foundation of experimental science.At the same time in clinical practice to erythropoietin treatment of retinal ischemia patients combined with fuming decoction,the clinical curative effect research.Methods 1 to 82 who were not infected with any disease of purebred white rabbits as the experimental object,which were randomly divided into 3 groups,normal group,injury group and treatment group with 10 rats in each group,36,36;injury group and treatment group two groups according to different reperfusion time,each divided into injury 1/4d,1/2d,1D,2D,4D,8D group,a total of 6 groups,each group of each 6 rabbits.2 all of the rabbits were treated with modeling and treatment of the right eye,except for the normal group of 10 rabbits,the model of retinal ischemia reperfusion injury was established by using anterior chamber perfusion to increase intraocular pressure.The normal animal at the ear part of the intravenous injection of 0.9%NaCl,dose of 2.0ml/kg,once a day;after modeling in injury group,from the ear part of the intravenous injection of 0.9%NaCl,the same dose of normal group;the treatment group in the model,by gavage in mice by intragastric injection of fuming Decoction,rabbits 5min after ischemia to the injection of rhEPO(1000U/k).3 after injury 1/4d,1/2d,1D,2D,4D,8D,from the ear edge of intravenous injection of 02 ml of 10 ml,immediately after the death of the eye,the embedded section.4 HE staining was used to observe and analyze the structure,each time the animal retinal changes;analysis of retinal cell Bcl-2 and Bax levels by immunohistochemistry;changes of nerve cells in the retina were observed by TUNEL method;using Fluorogold retrograde tracer method to evaluate the survival rate of RGCs.All the experimental data were statistically analyzed,and the statistical software was processed by SPSS 18.0.Clinical research in Ophthalmology,Harbin 242 hospital diagnosed 60 patients with central retinal artery occlusion as the research object,according to the conventional treatment group,the study group based on routine treatment,in the treatment of combined cataract by erythropoietin drink.The study group,the control group of 30 patients,the number of eyes need to be treated for only 36,only 35,to improve the strength of patients to check.Results 1.HE staining:(1)the number of ganglion cells in the injured group was lower than that in the normal group.The injury group,1/4d injury after mild retinal edema,and with the extension of injury,edema is more serious,cell gap increased,the volume increased,vacuoles obviously;edema after 1D,the most serious edema then 2d-4d,edema gradually reduced,the cell gap narrows gradually fade;edema 8D after the return to normal cell gap.(2)the number of ganglion cells in the treatment group was lower than that in the injured group,and the retinal edema was more significant than that in the injury group.2 immunohistochemical analysis:(1)normal group:Bcl-2 and Bax protein were negative.(2)injury group:1/4d after injury,Bcl-2 and Bax protein in the retina of the minority was positive in ganglion cell gap;with increasing injury,Bcl-2,positive expression of Bax protein is more obvious,the number of 1D positive expression in most,a positive kernel layer,outer nuclear layer was negative;the number of 2d-4d positive expression decreased,most were expressed in the ganglion cell gap;8D after injury,all negative expression.(3)the expression of Bcl-2 and Bax in the treatment group was similar to that in the injury group.Compared with 1/4d,1/2d,1D,2D,4D,8D after injury,the expression level of Bcl-2 in the treatment group was significantly higher than that in 8D group.The level of Bax was significantly lower in the treatment group than in the injury group(P<0.05).3.TUNEL analysis showed that:(1)normal group:no apoptosis of nerve cells.(2)injury group:1/4d after injury,a nerve cell apoptotic,with time prolonging,the apoptosis was gradually serious,positive increase in the number of 1D after injury,the positive expression rate rose to the highest,2D decreased,decreased significantly after 3D,5D positive expression after all disappear.(3)in the treatment group,the apoptosis of nerve cells was similar to that in the injury group,but the positive expression rate of apoptotic cells was significantly lower than that in the injury group.There was no significant difference between the injury group and the treatment group in the 1/4d,1/2d,1D,2D,4D,5D(P<0.01)after injury,and there was no significant difference between the two groups(P>0.05).The survival rate of 4.RGCs:(1)the survival rate of RGCs in the normal group was respectively at the time of 63%,66%,3 8%,25%,15%,respectively.(2)the survival rate of RGCs in the injury group was respectively at the time of 82%,83%,50%,37%,22%,respectively.(3)the survival rate of RGCs in the treatment group was:95%,94%,73%,53%,and 25%,respectively.There was a significant difference between the three groups(P<0.05).Clinical trials,comparative vision detection before and after treatment in two groups:the study group after treatment,the average sensitivity level had increased than before treatment,and the study group increased after treatment was significantly higher than the control group,two groups have significant difference(P<0.05);the study group after treatment,the average defect degree than before treatment decreased significantly and after treatment significantly higher than the control group,compared with significant difference(P<0.05).There was no difference in IOP between the two groups before and after treatment.There were differences in IOP and the difference of IOP before and after treatment(P<0.05).There was no difference in amplitude of B wave before treatment between the two groups.The amplitude of B wave and difference of B wave amplitude before and after treatment were different(P<0.05).After treatment,the study group cured,markedly effective,5 cases were invalid,16 cases,5 cases,4 cases,the total efficiency of 86.7%;control group of 2 cases were cured,11 cases markedly effective,7 cases effective,10 cases ineffective,the total efficiency of 66.7%,significantly less than the study group,there was statistical significant differences(P<0.05).Conclusion rhEPO combined with fuming Decoction on retinal ischemia reperfusion injury has obvious clinical curative effect of treatment,can help to maintain the normal structure of the retina;fuming decoction has obvious inhibitory effect on the apoptosis of retinal nerve cells,the main reason is that Bcl-2,Bax and protein expression level. |
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