上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (12): 1697-.doi: 10.3969/j.issn.1674-8115.2011.12.008

• 论著（基础研究） • 上一篇    下一篇

缺血再灌注损伤后大鼠视网膜细胞凋亡与低氧诱导因子1α表达的关系

袁海虹1, 周 薇2, 包辉英1, 谭攀攀2, 祝肇荣3, 吴国忠1   

  1. 1.上海医药高等专科学校药理学教研室, 上海 201318； 2.上海交通大学 医学院药理学教研室, 上海 200025； 3.上海交通大学 医学院附属新华医院眼科, 上海 200092
  • 出版日期:2011-12-28 发布日期:2012-01-04
  • 通讯作者: 吴国忠, 电子信箱: guozhongwu@hotmail.com。
  • 作者简介:袁海虹（1977—）, 女, 讲师, 硕士；电子信箱: yuanhaihong@126.com。
  • 基金资助:

    上海市自然科学基金（11ZR1419300）；上海交通大学医学院自然科学基金（09XJ21079）；上海市教委科研创新项目(10YZ241)

Relationship between cell apoptosis and expression of hypoxia-induced factor-1alpha in retinal cells after ischemia-reperfusion injury in rats

YUAN Hai-hong1, ZHOU Wei2, BAO Hui-ying1, TAN Pan-pan2, ZHU Zhao-rong3, WU Guo-zhong1   

  1. 1.Laboratory of Pharmacology, Shanghai Institute of Health Sciences, Shanghai 201318, China；2.Laboratory of Pharmacology, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China；3.Department of Ophthalmology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200092, China
  • Online:2011-12-28 Published:2012-01-04
  • Supported by:

    Natural Science Foundation of Shanghai, 11ZR1419300；Natural Science Foundation of Shanghai Jiaotong University School of Medicine, 09XJ21079；Innovation Program of Shanghai Education Committee, 10YZ241

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摘要：

目的 研究缺血再灌注损伤后大鼠视网膜细胞凋亡与低氧诱导因子1α(HIF-1α)的表达变化并探讨两者的相关性。方法 雄性SD大鼠随机分为正常对照组和缺血再灌注损伤1 d、3 d和5 d组(n=6)。眼前房灌注生理盐水调节大鼠眼压至110 mmHg并持续50 min，制作实验性视网膜缺血再灌注损伤模型。制备视网膜切片，测量内网层(IPL)、内核层(INL)厚度及节细胞层（GCL）细胞数；采用DNA原位末端标记(TUNEL)法检测视网膜凋亡细胞；免疫组织化学染色观察损伤后不同时间HIF-1α在视网膜细胞中的表达；采用RT-PCR检测HIF-1α mRNA的表达。结果 缺血再灌注损伤1 d、3 d和5 d组大鼠的IPL和INL厚度显著小于正常对照组（P<0.01），GCL细胞少于正常对照组（P<0.05）。视网膜细胞凋亡位于GCL；缺血再灌注损伤1 d、3 d和5d组的细胞凋亡率均明显高于正常对照组，差异有统计学意义（P<0.01）。缺血再灌注损伤1 d、3 d、5 d组视网膜GCL的HIF-1α蛋白表达阳性细胞比例分别为（47.88±14.71）%、（50.28±13.11）%、（43.09±10.04）%，与视网膜细胞凋亡率呈正相关（r=0.953）。缺血再灌注损伤3 d组视网膜细胞HIF-1α mRNA表达明显高于正常对照组(P< 0.05)。结论 缺血再灌注后大鼠视网膜细胞凋亡与HIF-1α的表达均增加，两者呈正相关；HIF-1α的表达可能参与视网膜细胞凋亡。

关键词: 缺血再灌注, 视网膜, 低氧诱导因子1α, 凋亡

Abstract:

Objective To investigate the changes of cell apoptosis and expression of hypoxia-induced factor-1α (HIF-1α) in retinal cells after ischemiareperfusion injury in rats, and explore the relationship between them. Methods Male SD rats were randomly divided into normal control group, 1 d after ischemia-reperfusion injury group, 3 d after ischemia-reperfusion injury group and 5 d after ischemia-reperfusion injury group (n=6). Retinal ischemia-reperfusion injury model was induced by perfusing saline into anterior chamber of eye to generate the intraocular pressure of 110 mmHg for 50 min. Retinal sections were prepared, the thickness of inner plexiform layer (IPL) and inner nuclear layer (INL) and the number of cells in ganglion cell layer (GCL) were measured, the retinal cell apoptosis was determined by TUNEL, the expression of HIF-1α in retinal cells was observed with immunohistochemical staining, and the expression of HIF-1α mRNA at different time points after injury was detected by RTPCR. Results Compared with normal control group, the thickness of IPL and INL was significantly lower (P<0.01), and the number of cells in GCL was significantly smaller in 1 d after ischemia-reperfusion injury group, 3 d after ischemia-reperfusion injury group and 5 d after ischemia-reperfusion injury group (P<0.05). Apoptosis of retinal cells occurred in GCL, and the apoptosis rates in 1 d after ischemia-reperfusion injury group, 3 d after ischemiareperfusion injury group and 5 d after ischemia-reperfusion injury group were significantly higher than that in normal control group (P<0.01). The rates of positive expression of HIF-1α in GCL of retina in 1 d after ischemia-reperfusion injury group, 3 d after ischemia-reperfusion injury group and 5 d after ischemia-reperfusion injury group were （47.88±14.71）%,（50.28±13.11）% and (43.09±10.04)% respectively, which were in line with the tendency of apoptosis of retinal cells (r=0.953). The expression of HIF-1α mRNA in 3 d after ischemia-reperfusion injury group was significantly higher than that in normal control group (P<0.05). Conclusion Both cell apoptosis and expression of HIF-1α in retinal cells increase after ischemiareperfusion injury in rats, and there is a positive correlation between them. The expression of HIF-1α may play an important role in retinal cell apoptosis.

Key words: ischemia reperfusion, retina, hypoxia-induced factor-1α, apoptosis