Abstract：
OBJECTIVE: To investigate the possible roles of Interleukin 17A (IL-17A) and IL-17A neutralizing antibodies (IL-17Ab) in glaucoma and the potential mechanisms.
METHODS: The two glaucoma animal models, chronic ocular hypertension (COH) and N-methyl-D-aspartate (NMDA)-induced retinal ganglion cell (RGC) damage, were established and treated with intravitreal injection of IL-17A or IL-17Ab. Intraocular pressure (IOP) was measured by a rebound tonometer. The retina and RGC injury were evaluated by HE staining, TUNLE assay and Brn3a immunofluorescence staining. The frequency of IL-17A+CD4+T cells in peripheral blood was detected by flow cytometry. The expression of glial fibrillary acidic protein (GFAP) was detected by immunofluorescence staining, Western Blot and qPCR in retina. The RNA and protein expression of Act1/TRAF6/NF-κB were detected by Western Blot and qPCR in retina.
RESULTS: The expression of IL-17A increased in glaucoma models. After intravitreal injection of IL-17A, in the retina, the number of RGCs decreased, the apoptosis of RGCs increased, the Müller cell gliosis was more obvious. In addition, peripheral inflammation aggravated. Whereas the intravitreal injection of IL-17Ab alleviated the relevant manifestations and peripheral inflammation, reduced the gliosis of Müller cells. In the COH model, IOP increased after the injection of IL-17A, while the intravitreal injection of IL-17Ab led to a decrease in IOP. Furthermore, IL-17A promotes the apoptosis of RGCs by binding to IL-17A receptor, activating Act1/TRAF6/NF-κB pathways.
CONCLUSIONS: IL-17A plays a role in and aggravates RGC damage in glaucoma. IL-17Ab can neutralize the pro-inflammatory effect of IL-17A and have a protective function in glaucoma. These findings reveal the importance of IL-17A in the pathogenesis of glaucoma, which will shed light on a novel direction for the prevention and treatment of glaucoma, and also provide a reference for further research on other retinal diseases.
METHODS: The two glaucoma animal models, chronic ocular hypertension (COH) and N-methyl-D-aspartate (NMDA)-induced retinal ganglion cell (RGC) damage, were established and treated with intravitreal injection of IL-17A or IL-17Ab. Intraocular pressure (IOP) was measured by a rebound tonometer. The retina and RGC injury were evaluated by HE staining, TUNLE assay and Brn3a immunofluorescence staining. The frequency of IL-17A+CD4+T cells in peripheral blood was detected by flow cytometry. The expression of glial fibrillary acidic protein (GFAP) was detected by immunofluorescence staining, Western Blot and qPCR in retina. The RNA and protein expression of Act1/TRAF6/NF-κB were detected by Western Blot and qPCR in retina.
RESULTS: The expression of IL-17A increased in glaucoma models. After intravitreal injection of IL-17A, in the retina, the number of RGCs decreased, the apoptosis of RGCs increased, the Müller cell gliosis was more obvious. In addition, peripheral inflammation aggravated. Whereas the intravitreal injection of IL-17Ab alleviated the relevant manifestations and peripheral inflammation, reduced the gliosis of Müller cells. In the COH model, IOP increased after the injection of IL-17A, while the intravitreal injection of IL-17Ab led to a decrease in IOP. Furthermore, IL-17A promotes the apoptosis of RGCs by binding to IL-17A receptor, activating Act1/TRAF6/NF-κB pathways.
CONCLUSIONS: IL-17A plays a role in and aggravates RGC damage in glaucoma. IL-17Ab can neutralize the pro-inflammatory effect of IL-17A and have a protective function in glaucoma. These findings reveal the importance of IL-17A in the pathogenesis of glaucoma, which will shed light on a novel direction for the prevention and treatment of glaucoma, and also provide a reference for further research on other retinal diseases.
摘要：
目的：探讨白细胞介素17A(IL-17A)和IL-17A中和抗体(IL-17Ab)在青光眼发病中的可能作用及可能的机制。
方法：两种青光眼动物模型，慢性高眼压（COH）和N-甲基-D-天冬氨酸（NMDA）诱导的视网膜神经节细胞（RGC）损伤，建立并用玻璃体内注射IL-17A或IL-17Ab治疗。通过回弹眼压计测量眼压(IOP)。HE染色评价视网膜和RGC损伤,TUNLE测定和Brn3a免疫荧光染色。流式细胞术检测外周血IL-17A+CD4+T细胞的频率。免疫荧光染色检测胶质纤维酸性蛋白(GFAP)的表达,视网膜中的Western印迹和qPCR。WesternBlot和qPCR检测视网膜中Act1/TRAF6/NF-κB的RNA和蛋白表达。
结果：青光眼模型中IL-17A的表达增加。玻璃体内注射IL-17A后，在视网膜上，RGC的数量减少，RGCs的凋亡增加，Müller细胞胶质增生更为明显。此外,周围炎症加重。而玻璃体内注射IL-17Ab减轻了相关表现和外周炎症，减少了Müller细胞的神经胶质增生。在COH模型中，注射IL-17A后眼压升高,而玻璃体内注射IL-17Ab导致IOP降低。此外，IL-17A通过与IL-17A受体结合促进RGCs凋亡,激活Act1/TRAF6/NF-κB通路。
结论：IL-17A在青光眼的RGC损害中起作用并加重。IL-17Ab可以中和IL-17A的促炎作用，并在青光眼中具有保护功能。这些发现揭示了IL-17A在青光眼发病机制中的重要性。这将揭示青光眼预防和治疗的新方向，为其他视网膜疾病的进一步研究提供参考。
方法：两种青光眼动物模型，慢性高眼压（COH）和N-甲基-D-天冬氨酸（NMDA）诱导的视网膜神经节细胞（RGC）损伤，建立并用玻璃体内注射IL-17A或IL-17Ab治疗。通过回弹眼压计测量眼压(IOP)。HE染色评价视网膜和RGC损伤,TUNLE测定和Brn3a免疫荧光染色。流式细胞术检测外周血IL-17A+CD4+T细胞的频率。免疫荧光染色检测胶质纤维酸性蛋白(GFAP)的表达,视网膜中的Western印迹和qPCR。WesternBlot和qPCR检测视网膜中Act1/TRAF6/NF-κB的RNA和蛋白表达。
结果：青光眼模型中IL-17A的表达增加。玻璃体内注射IL-17A后，在视网膜上，RGC的数量减少，RGCs的凋亡增加，Müller细胞胶质增生更为明显。此外,周围炎症加重。而玻璃体内注射IL-17Ab减轻了相关表现和外周炎症，减少了Müller细胞的神经胶质增生。在COH模型中，注射IL-17A后眼压升高,而玻璃体内注射IL-17Ab导致IOP降低。此外，IL-17A通过与IL-17A受体结合促进RGCs凋亡,激活Act1/TRAF6/NF-κB通路。
结论：IL-17A在青光眼的RGC损害中起作用并加重。IL-17Ab可以中和IL-17A的促炎作用，并在青光眼中具有保护功能。这些发现揭示了IL-17A在青光眼发病机制中的重要性。这将揭示青光眼预防和治疗的新方向，为其他视网膜疾病的进一步研究提供参考。
