| Objectives:Protective effect of Lycium barbarum polysaccharide on rat retina ischemia-reperfusion injury,The possible mechanism of the protective effect of Lycium barbarum polysaccharide on rat retina ischemia-reperfusion injury,providing theoretical basis and new diagnosis and treatment for the treatment of retinal ischemia-reperfusion injury.Methods:(1)55 healthy adult male Sprague-Dawley rats were used as experimental subjects,and their body weight ranged from 200 to 300 g.They were purchased from the Institute of Animal Science of Nanchang University.The digital table method was randomly divided into blank control group,sham operation group,and ischemiareperfusion group.Ischemia-reperfusion model+ sputum polysaccharide group referred to as LBP drug group,sham operation group 5,the other two groups each group 25,each component a total of 5 time points(6h,12 h,24h,48 h,72h,),5 at each time point,at room temperature,give enough food and water,12 h light,adapt to sexual life for 7 days under 12 h night.(2)SD male rats in the LBP drug group were intragastrically treated with4ml/100 g sputum polysaccharide solution(once a day for one week),and the model group SD male rats and sham operation group were treated with 4ml/100 g 0.9%sodium chloride solution.(once a day,for one week in a row).(3)A model of retinal ischemia-reperfusion injury was made by clamping the right common carotid artery of the rat with arterial clip.The right common carotid artery was clamped in the model group and the right common carotid artery of the LBP drug group(clamped for 1 hour).The sham operation group was only operated for 1 hour without exposing the right common carotid artery.The incision was sutured layer by layer after 1 hour.(4)SD rats were treated at 6h,12 h,24h,48 h and 72 h after ischemia-reperfusion.SD rats were sacrificed by intraperitoneal injection of 10% chloral hydrate solution under deep anesthesia.Eye stains were made by HE staining.The changes of morphological structure of the retina were observed under microscope.The expression of Caspase-3,Bcl-2 and Bax protein in the retina of each group were detected by immunohistochemical staining.Results:(1)HE staining results: 6 hours after retinal ischemia-reperfusion,mild retinal edema,slight increase in retinal thickness,edema of nerve fiber layer and inner plexiform layer,12,24 hours after retinal ischemia-reperfusion,severe retinal edema The number of ganglion cells decreased,the inner nuclear layer was thin,and the cells were loosely arranged;48 hours of edema disappeared,mainly showing that the number of retinal ganglion cells continued to decrease,the inner nuclear layer was further thinned,and vacuolization was observed;72 hours of retinal thinning The number of retinal cells is reduced.Compared with the model group(RIRI group),the retinal tissue edema was relieved at 6h and 12 h,24h in the drug group(LBP group);the ganglion cells were less reduced in the model group at 24 h,48h and 72 h,and there was no significant change in retinal thickness.The layers of the retina of the drug group are basically ordered.(2)The results of immunohistochemistry showed that the number of Caspase-3positive cells in the sham operation group was(103.12±12.97)·mm-2.The number of Caspase-3 positive cells in the model group increased at 6 h after perfusion,and the number of positive cells increased.(559.03±38.25)mm-2,24 h reached a high level,the number of positive cells was(1528.12 ± 52.74)mm-2,and then gradually decreased,LBP drug group retinal active Caspase-3 positive at each time point The number of cells was significantly lower than that of the ischemia-reperfusion group,and the difference was statistically significant(all P<0.05).A large number of Bcl-2positive cells were observed in the sham-operated group.Bcl-2 positive cells began to decrease at 6 h after RIRI in the ischemia-reperfusion group,and continued to decrease after 12 h and decreased to a lower level at 24 h.At each time point,the number of Bcl-2 positive cells in the drug group was significantly higher than that inthe ischemia-reperfused retina,and the difference was statistically significant(all P<0.05).Bax positive cells were less expressed in the sham-operated rats;in the ischemia-reperfusion group,Bax-positive cells were observed in the ganglion cell layer and the inner nuclear layer 6 hours after RIRI,and reached a higher level at 24 hours.And began to decrease in 48 hours.At each time point,the Bax positive cells in the drug group were significantly less than the ischemia-reperfusion group,and the differences were statistically significant(all P<0.05).Conclusions:Lycium barbarum polysaccharide has protective effects on retinal damage caused by retinal ischemia-reperfusion injury.Its protection is to inhibit apoptosis in retinal ischemia-reperfusion injury,which is related to up-regulation of Bcl-2 and down-regulation of Bax to Caspase-3. |
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