AH samples were collected from 20 patients during cataract surgeries (controls: n = 10, age = 70.3 ± 9.742; glaucoma: n = 10, age: 66.5 ± 8.073) in Shanghai East Hospital, an affiliate of Tongji University, between September 2018 and March 2019 and analyzed in duplicate by Luminex cytokine polystyrene color bead-based multiplex assay. Retinas from female Sprague-Dawley rats (n = 6) were harvested ex vivo and cultured with or without 60 mmHg of hydrostatic pressure for 24 hours. Retinal ganglion cells (RGCs) were quantified using Brn3a staining. Cytokines in the retina and culture medium were analyzed by rat cytokine array (Abcam).
At baseline, patients with primary angle closure glaucoma (PACG) have significantly lower levels of IL-6 and IP-10 and a higher level of PDGF-BB in their AH, compared to the controls. Postoperatively, patients with PACG have significantly higher levels of IL-1ra, IL-13, and MIP-1α and a lower level of IL-6. Elevated hydrostatic pressure led to significant RGC loss in the retina, ex vivo, as well as the upregulation of ciliary neurotrophic factor (CNTF), IL-6, IL-10, IL-4, and TIMP-1 alongside the downregulation of PDGF-AA, MMP-8, TNF-α, and IFN-γ. Furthermore, eight cytokines were detected as being downregulated in the culture medium, including PDGF-AA, MMP-8, and IL-4.
Proinflammatory cytokines showed changes in both AH and ex vivo. Further studies are needed on the role of these cytokines and their corresponding signaling pathways in both neurodegeneration and glaucoma.
方法:收集20例白内障手术患者的AH样本(对照组:n=10,年龄=70.3±9.742;青光眼:n=10,年龄:66.5±8.073)。在上海东方医院,同济大学的附属机构,2018年9月至2019年3月,并通过基于Luminex细胞因子聚苯乙烯彩珠的多重测定进行一式两份分析。离体收获来自雌性Sprague-Dawley大鼠(n=6)的视网膜,并在有或没有60mmHg静水压力的情况下培养24小时。使用Brn3a染色定量视网膜神经节细胞(RGC)。通过大鼠细胞因子阵列(Abcam)分析视网膜和培养基中的细胞因子。
结果:在基线时,原发性闭角型青光眼(PACG)患者的AH中IL-6和IP-10水平明显较低,PDGF-BB水平较高,与对照组相比。术后,PACG患者的IL-1ra水平明显较高,IL-13和MIP-1α以及较低水平的IL-6。静水压力升高导致视网膜中的RGC明显丢失,离体,以及睫状神经营养因子(CNTF)的上调,IL-6,IL-10,IL-4和TIMP-1伴随PDGF-AA的下调,MMP-8,TNF-α,和IFN-γ。此外,在培养基中检测到8种细胞因子被下调,包括PDGF-AA,MMP-8和IL-4。
结论:促炎细胞因子在AH和离体中均显示出变化。需要进一步研究这些细胞因子及其相应的信号通路在神经变性和青光眼中的作用。