白细胞介素17A在视神经损伤模型小鼠神经再生中的作用

Interleukin-17A facilitates optic nerve regeneration in a mouse model of optic nerve crush

背景:实验证明在角膜损伤模型中,白细胞介素17A参与的炎症反应在角膜神经再生过程中发挥着重要作用。目的:进一步验证白细胞介素17A对视神经损伤模型小鼠神经再生的作用。方法:构建白细胞介素17A过表达腺病毒载体(AV-IL-17A-GFP),构建白细胞介素17A表达载体,包装并纯化腺病毒,以绿色荧光蛋白空载体包装的腺病毒为对照组(AV-GFP对照组);用病毒感染PC12细胞并通过ELISA检测白细胞介素17A表达效率,采用显微注射方法将病毒注射到小鼠玻璃体内,利用免疫荧光检测视神经中白细胞介素17A的表达,构建小鼠视神经损伤模型,检测受损视神经再生情况。结果与结论:(1)与AV-GFP对照组相比,ELISA检测发现AV-IL-17A-GFP组中白细胞介素17A在PC12细胞中高度表达;(2)免疫荧光染色表明白细胞介素17A在小鼠视神经节细胞和视神经中有效表达,AV-IL-17A-GFP组中再生的视神经纤维数量显著多于对照组,且长度显著长于对照组(P<0.05);(3)结果说明,白细胞介素17A腺病毒载体可在小鼠视神经节细胞和视神经中表达,并且能够促进受损视神经的再生。

BACKGROUND:Interleukin-17A has been shown to play an important role in nerve regeneration following corneal injury.OBJECTIVE:To investigate the effect of interleukin-17A on mouse nerve regeneration after optic nerve injury.METHODS:The interleukin-17A-overexpressed adenovirus vector(AV-IL-17A-GFP)was constructed and purified.Adenoviruses with green fluorescent protein(AV-GFP)served as control group.PC12 cell lines were transfected with adenoviruses and the expression efficiency of IL-17A were detected by ELISA.The adenovirus was intravitreally injected into the mouse vitreous body,and the expression level of IL-17A in the optic nerve was determined by immunofluorescence.Afterwards,a mouse model of optic nerve injury was established to observe the nerve regeneration.RESULTS AND CONCLUSION:ELISA results showed that IL-17A was highly expressed in the PC12 cell lines transfected with AV-IL-17A-GFP compared with the control group.Immunofluorescence results revealed that IL-17A was expressed in vivo successfully.There was a significant increase in the number of regenerated axons in the AV-IL-17A-GFP group compared with the control group(P<0.05).To conclude,AV-IL-17A-GFP can be expressed in the mature neurons in vivo and promote the axon regeneration after optic nerve crush injury in mice.