Eye Institute, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China.
Medical School of Nantong University, Nantong, Jiangsu, China.
Invest Ophthalmol Vis Sci. 2024 Sep 3;65(11):21. doi: 10.1167/iovs.65.11.21.
This study aims to investigate the relationship among STRA6, circadian rhythm, and choroidal neovascularization (CNV) formation, as well as the regulatory mechanism of STRA6 in CNV under circadian rhythm disturbances.
C57BL/6J male mice (aged 6 weeks) were randomly divided into control and jet lag groups (using a time shift method every 4 days to disrupt the molecular clock's capacity to synchronize with a stable rhythm). A laser-induced CNV model was established in both the control and the jet lag group after 2 weeks of jet lag. The size of CNV lesions and vascular leakage were detected by morphological and imaging examination on the seventh day post laser. STRA6 was screened by full transcriptome sequencing. Bioinformatics analysis was conducted to assess the variation and association of STRA6 in the GSE29801 dataset. The effects of STRA6 were evaluated both in vivo and in vitro. The pathway mechanism was further elucidated and confirmed through immunofluorescence of paraffin sections and Western blotting.
The disturbance of circadian rhythm promotes the formation of CNV. Patients with age-related macular degeneration (AMD) exhibited higher levels of STRA6 expression compared to the control group, and STRA6 was enriched in pathways related to angiogenesis. In addition, CLOCK and BMAL1, which are initiators that drive the circadian cycle, had regulatory effects on STRA6. Knocking down STRA6 reversed the promotion of CNV formation caused by circadian rhythm disturbance in vivo, and it also affected the proliferation, migration, and VEGF secretion of RPE cells without circadian rhythm in vitro, as well as impacting endothelial cells. Through activation of the JAK2/STAT3/VEGFA signaling pathway in unsynchronized RPE cells, STRA6 promotes CNV formation.
This study suggests that STRA6 reduces CNV production by inhibiting JAK2/STAT3 phosphorylation after circadian rhythm disturbance. The results suggest that STRA6 may be a new direction for the treatment of AMD.
本研究旨在探讨 STRA6、昼夜节律和脉络膜新生血管（CNV）形成之间的关系，以及 STRA6 在昼夜节律紊乱下对 CNV 的调控机制。
将 6 周龄 C57BL/6J 雄性小鼠随机分为对照组和时差组（每 4 天进行一次时间转移以破坏分子钟与稳定节律同步的能力）。在时差 2 周后，两组均建立激光诱导的 CNV 模型。在激光后第 7 天通过形态学和影像学检查检测 CNV 病变和血管渗漏的大小。通过全转录组测序筛选 STRA6。通过 GSE29801 数据集进行生物信息学分析，评估 STRA6 的变化和相关性。在体内和体外评估 STRA6 的作用。通过石蜡切片免疫荧光和 Western blot 进一步阐明和证实通路机制。
昼夜节律紊乱促进 CNV 的形成。与对照组相比，年龄相关性黄斑变性（AMD）患者的 STRA6 表达水平更高，并且 STRA6 富集在与血管生成相关的途径中。此外，作为驱动昼夜节律周期的启动子的 CLOCK 和 BMAL1 对 STRA6 具有调节作用。体内敲低 STRA6 逆转了昼夜节律紊乱引起的 CNV 形成的促进作用，并且它还影响了体外无昼夜节律的 RPE 细胞的增殖、迁移和 VEGF 分泌，以及影响内皮细胞。通过激活非同步 RPE 细胞中的 JAK2/STAT3/VEGFA 信号通路，STRA6 促进 CNV 的形成。
本研究表明，STRA6 通过抑制昼夜节律紊乱后 JAK2/STAT3 磷酸化来减少 CNV 的产生。结果表明，STRA6 可能是治疗 AMD 的一个新方向。
