摘要
目的观察低氧预适应(hypoxic preconditioning,HPC)后视网膜缺血再灌注损伤(retinal ischemia-reperfusion injury,RIRI)大鼠视网膜促红细胞生成素(erythropoietin,EPO)蛋白及活性Caspase-3蛋白表达的动态变化,探讨HPC预防RIRI的机制,为RIRI的防治提供有效的治疗方案。方法取健康Sprague Dawley成年大鼠随机分为正常对照组(CON组),RIRI组与HPC+缺血再灌注损伤组(HPC组,HPC后采用高眼压法制成RIRI模型),每组根据RIRI时间分为6h、12h、24h及72h4个亚组。采用免疫组织化学法动态观察HPC对RIRI大鼠视网膜细胞EPO蛋白、活性Caspase-3蛋白表达的影响。结果 CON组大鼠视网膜仅见极少量的EPO+细胞;RIRI后6h,HPC组EPO+细胞数开始增加;RIRI后12h,HPC组EPO+细胞数(171±21)mm-2进一步增加,显著高于RIRI组[(149±20)mm-2,P<0.05];RIRI后24h,HPC组EPO+细胞数(287±24)mm-2达较高水平并显著高于RIRI组[(238±22)mm-2,P<0.01];RIRI后72h,HPC组EPO+细胞数开始下降,但仍显著高于RIRI组(P<0.05)。RIRI后6h,RIRI组活性Caspase-3+细胞数(60±5)mm-2开始增加,显著多于HPC组[(53±5)mm-2,P<0.05];RIRI后12h,RIRI组与HPC组活性Caspase-3+细胞数进一步增加,HPC组活性Caspase-3+细胞显著少于RIRI组(P<0.01);RIRI后24h,RIRI组与HPC组活性Caspase-3+细胞数达较高水平,HPC组活性Caspase-3+细胞数(578±26)mm-2显著少于RIRI组[(624±25)mm-2,P<0.01];RIRI后72h,HPC组与RIRI组活性Caspase-3+细胞数均开始下降,HPC组Caspase-3+细胞数仍少于RIRI组(P<0.05)。结论 HPC可促进RIRI大鼠视网膜细胞EPO蛋白的表达,减轻活性Caspase-3蛋白的表达,从而减轻细胞凋亡,具有神经保护作用。
Objective To observe the dynamic changes of erythropoietin （EPO） and active Caspase-3 protein expression in the rats with retinal ischemia-reperfusion in- jury （RIRI） after hypoxia preconditioning （HPC）, and explore the mechanism of HPC preventing RIRI,thus provide effective therapeutic schedule for the prevention of RIRI. Methods The healthy Sprague Dawley adult rats were randomly divided into the nor- mal control （CON） group,RIRI group and HPC group （RIRI models were established in the rats of this group after giving HPC）, the groups were sub-divided into 6 hours, 12 hours,24 hours and 72 hours group based on the time of RIRI. The effects of HPC on EPO and active Caspase-3 protein expression were examined by immunohistochemical method. Results The EPO ~ cells were seldom observed in retina of CON group. Six hours after RIRI,the EPO ＋ cells began to increase in the HPC group;12 hours after RI- RI,the EPO ＋ cells （171±21 ）ram-2 increased in the HPC group,which was significantly higher than that in the RIRI group [ （ 149±20 ） mm - 2,p 〈 0.05 ] ; 24 hours after RIRI, the EPO＋ cells （287 ±24）mm^-2： increased to the highest level in the HPC group,which were significantly higher than that of the RIRI group [ （238±22） mm^-2 ,P 〈 0.01 ] ;72 hours after RIRI, the EPO ~ cells in the HPC group began to drop, which were still higher than those in the RIRI group （P 〈 0.05 ）. Six hours after RIRI, the active Caspase-3 ^＋ cells （50±5 ）mm^-2 of the RIRI group increased,which were significantly higher than that in the HPC group E （53±5 ）mm - 2,p 〈 0. 05 ] ; 12 hours after RIRI, the active Caspase-3 in- creased in both the HPC group and RIRI group, and the active Caspase-3^＋ cells in the HPC group were significantly lower than that in the RIRI group（P 〈0.01 ） ;24 hours af- ter RIRI,the active Caspase-3 increased to the highest level in both the RIRI group and HPC group,and the active Caspase-3^＋ cells in the HPC group （578± 25）mm-2 were lower than that in the RIRI group [ （624±25 ） ram-2 ,p 〈 0.01 ] ;72 hours after the RIRI, the active Caspase-3 ＋ cells began to decease in both the RIRI and HPC group, and the active Caspase-3 ＊ cells in the RIRI group were still higher than that in the HPC group（P 〈 0.05）. Conclusion HPC can promote the expression of EPO protein and reduce the expression of the active Caspase-3 protein, thus alleviate the apoptosis of retinal cells with neuro-protective effects.
出处
《眼科新进展》
CAS
北大核心
2013年第1期5-8,共4页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:81000268)
山东省自然科学基金项目(编号:ZR2010HQ037)
山东省教育厅课题(编号:J11LF72)~~
