摘要
目的:探讨磷酰肌醇-3激酶(phosphatidylinositol-3 kinase,PI3K)途径在大鼠视网膜缺血再灌注后对低氧诱导因子(HIF-1α)表达的影响。方法:将磷酰肌醇-3激酶(PI3K)特异抑制剂Wortmannin(WT)注入大鼠玻璃体内,抑制PI3K活性为A组,玻璃体内注射等量林格液作为B组,空白对照组为C组,采用前房灌注生理盐水升高大鼠眼压到110mm Hg(1 kPa=7.5 mm Hg)持续1 h的方法制作实验性视网膜缺血再灌注损伤模型,于再灌注后0、2、6、8、12、24、48 h取材,行免疫组织化学法染色观察HIF-1α在视网膜细胞的阳性表达;并应用RT-PCR来检测HIF-1αmRNA的表达。结果:视网膜缺血再灌注6、8、12、24、48 h,视网膜神经节细胞层及内核层细胞出现HIF-1α阳性表达且表达逐渐增加,12 h达到高峰,然后逐渐降低,预先玻璃体内注射PI-3K特异抑制剂Wortmannin(WT)的大鼠视网膜HIF-1α表达明显减弱,亦在12 h达到高峰,然后逐渐降低。结论:缺血再灌注后大鼠视网膜HIF-1α的表达是经过PI3K途径来实现,阻断PI3K途径,可以明显减少HIF-1α的表达,但从实验结果来看PI3K途径被阻断后并没有完全阻断HIF-1α的表达,由此猜测HIF-1α的表达可能存在多种途径。
Objective ： To investigate the expression of HIF1αin rats＇ retinal ischemia-reperfusion injury,after intravitreal injection idio-inhibitor wortmannin. Methods ：Experimental group （A）：PI3K idio-inhibitor wortmannin 0.1 ml was injected intravitreal;Experimental control group （B）; Ringer＇s solation 0. 1 ml was iniected intravitreal.The rat model of experimental retinal ischemia reperfusion injury was created by increasing the intraocular pressure to 110 mmHg （1 kPa= 7. 5 mm Hg） in rat eyes. At different times post ischemia,the immunohistochemical staining and RT- PCR were done to observe the expression of HIF-1α. Results：In experimental control group;HIF-1αwas expressed in the cells of retinal ganglion layer and inner nuclear layer at 6 hours post ischemia. The expression attained the highest level at the 12 h after retinal ischemia eperfusion then the expression was declined. In experimental group ：the expression of HIF-1αwere obviously decreased and attained the highest level at the 12 h then declined. In control group ：the expression was low,and the level was almost unchanged. Conclusion ： The expression of HIF-1αafter retinal ischemia-reperfusion injury in rat comes true through PI3K pathway. If we block PI3K pathway,the expression of HIF-1αwill be decreased. But from the result we can see that wortmannin can not block the expression completely, we may guess expression of HIF-1α be completed through many way.
出处
《中国误诊学杂志》
CAS
2008年第5期1019-1021,共3页
Chinese Journal of Misdiagnostics
