热休克蛋白72在兔视网膜缺血再灌注损伤中的表达及重组人碱性成纤维细胞生长因子对其表达的影响

The expression of HSP72 in rabbit retina with ischemia-reperfusion injury and the effect of rh-bFGF on its expression

目的研究兔视网膜缺血再灌注损伤(RIRI)后重组人碱性成纤维细胞生长因子(rh-bFGF)对其组织中热休克蛋白(HSP)72活化程度的影响。方法选取66只大耳白兔,随机分为3组:缺血再灌注模型组(RIRI组30只)、rh-bFGF治疗组(RIRI+rh-bFGF组30只)、正常假手术组(6只),RIRI组、RIRI+rh-bFGF组又平均分为5个亚组(缺血再灌注后6、12、24、48、72h),每组6只大耳白兔。造模初期,RIRI组和rh-bFGF治疗组实验动物分别往玻璃体腔内注入相同剂量的0.9%氯化钠注射液和rh-bFGF溶液,并分别于制模后5个时间点处死动物摘取眼球,组织标本行苏木精-伊红(HE)染色及原位杂交。正常假手术组实验动物给予前房穿刺,无其他操作。常规HE染色后观察各组兔视网膜组织病理学变化,免疫组织化学法检测HSP72表达情况。结果正常假手术组兔视网膜各层次清晰、细胞平行排列且形态规则,视网膜神经节细胞呈单层排列、规则,无空泡变性。RIRI组兔视网膜出现高度水肿,层次紊乱、细胞结构疏松,视网膜神经节细胞出现空泡样变性,神经节细胞数目减少。RIRI+rh-bFGF组兔视网膜结构层次和细胞组织变化与RIRI组变化大致相同,但损害程度较其明显减轻。正常假手术组兔视网膜中没有发现HSP72;与RIRI组比较,RIRI+rh-bFGF组RIRI后5个时间点视网膜组织中HSP72的表达水平增高(P均<0.05)。结论 rh-bFGF对兔视网膜损伤有保护作用。

Objective To investigate the protective effect of recombinant human basic fibroblast growth factor（rh-bFGF）on retinal ischemia-reperfusion injury in rabbits.Methods The models of RIRI were made by elevating intraocular pressure.Sixty-six healthy rabbits were randomly divided into RIRI group,RIRI＋rh-bFGF group and sham-operation group.After the eyes in RIRI group and RIRI＋rh-bFGF group were established the ischemia-reperfusion injury models,saline was given to rabbits in RIRI group and rh-bFGF drug therapy was performed in RIRI＋rh-bFGF group.RIRI group and RIRI＋rh-bFGF group were subdivided into 6hours,12 hours,24hours,48 hours and 72 hours after reperfusion groups.Histopathological change was detected with light microscopy and immunohistochemical method was used to measure changes of HSP72 protein levels in retinal tissues.Results No apoptotic cells was found in sham-operation group.Apoptotic cells mainly appeared in the inner nuclear layer and the ganglion cell layer,and apoptotic cells went to peak at 24 hours after reperfusion.No expression of HSP72 positive cells were found in sham-operation group.In the RIRI group,the expression of HSP72 began to increase at 6hours after reperfusion.At 24 hours after reperfusion the expression reached the peak,and went on increasing with the reperfusion time prolonged,and began to decrease after 72 hours.The groups of RIRI＋rhbFGF had the same trend as the RIRI group in each index.RIRI＋rh-bFGF group being compared with RIRI group,HSP72 expression was significantly increased.There was significant difference between two groups（P〈0.05）.Conclusion The retina organization was injured by RIRI.HSP72 protein had taken part in the RIRI.rhbFGF could enhance the expression of HSP72 protein,so that the retina tissue was protected.