神经生长因子联合曲安奈德减轻缺血再灌注视网膜视神经节细胞损伤的研究

Effect of nerve growth factor combined with triamcinolone acetonide on retinal ganglion cell injury after ischemia-reperfusion

目的:观察神经生长因子(nerve growth factor, NGF)联合曲安奈德(triamcinolone acetonide, TA)对缺血再灌注大鼠视网膜损伤的作用,并探究其潜在的作用机制。方法:通过高眼压法建立大鼠视网膜缺血再灌注损伤模型;采用1 000 BU NGF或联合50μl TA玻璃体注射给药治疗大鼠。苏木素-伊红(hematoxylin eosin, HE)染色观察大鼠视网膜厚度、视网膜神经节细胞数量;末端标记法(TUNEL)检测细胞凋亡;蛋白质免疫印迹法(Western blotting, WB)实验检测半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase, Caspase)-3、B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X基因(Bax)、磷酸化p38分裂原激活的蛋白激酶(phosphorylation mitogen activated protein kinases, p-p38MAPK)和p38MAPK的蛋白表达;酶联免疫吸附实验(enzyme linked immunosorbent assays, ELISA)检测炎症因子白细胞介素(interleukin, IL)-1β、肿瘤坏死因子(tumor necrosis factor, TNF)-α和IL-6含量。结果:与正常对照组相比,模型对照组大鼠视网膜厚度、神经节细胞数目均显著减少,视网膜细胞凋亡率显著升高,Caspase-3、Bax蛋白表达显著升高,Bcl-2蛋白表达显著降低,IL-1β、TNF-α和IL-6含量均显著升高,p-p38MAPK蛋白表达显著降低;NGF或TA治疗均能显著减弱上述指标的变化,并且NGF联合TA的效果显著优于单独使用NGF。结论:NGF联合TA对视网膜缺血再灌注损伤的治疗效果优于单独使用NGF,其可能与抑制p38MAPK信号通路的活性有关。

Objective: To observe the effect of nerve growth factor(NGF) combined with triamcinolone acetonide(TA) on retinal injury in rats after ischemia-reperfusion, and to explore its potential mechanism. Methods: The rat model of retinal ischemia-reperfusion injury was established by high intraocular pressure method. The rats were treated with 1 000 BU NGF or combined with 50 μl TA intravitreal injection. Hematoxylin eosin(HE) staining was used to observe the retinal thickness and the number of retinal ganglions. Apoptosis was detected by terminal labeling(TUNEL). Western blotting(WB) was used to detect cysteine aspartate specific protein(Caspase)-3, B-cell lymphoma-2 gene(Bcl-2), Bcl-2 related X gene(Bax)and protein kinase activated by phosphorylated p38 mitogen(phosphorylation mitogen activated protein kinases, p-p38 MAPK) and p38 MAPK protein expression. Enzyme linked immunosorbent assays(ELISA) was used to detect the inflammatory factor interleukin(IL)-1β, tumor necrosis factor(TNF)-α. Results: Compared with the normal control group, the retinal thickness and the number of ganglion cells were significantly reduced, the apoptosis rate of retinal cells was significantly increased, the expression of Caspase-3 and Bax protein was significantly increased, the expression of Bcl-2 protein was significantly decreased, and the expression of IL-1β was significantly decreased, TNF-α and IL-6 were increased significantly, and the expression of p-p38 MAPK protein was decreased significantly in the model control group. NGF or combined with TA treatment could significantly reduce the changes of the above indexes, and the effect of NGF combined with TA was significantly better than that of NGF alone. Conclusion: The therapeutic effect of NGF combined with TA on retinal ischemia-reperfusion injury is better than that of NGF alone. Its potential mechanism may be related to the inhibition of p38 MAPK signal pathway.