Jin Zhong, Guo Qiru, Wang Zheng, Wu Xiao, Hu Wangming, Li Jiali, Li Hongfei, Zhu Song, Zhang Haidi, Chen Zixian, Xu Huan, Shi Liangqin, Yang Lan, Wang Yong
College of Basic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Chengdu University of Traditional Chinese Medicine, Hospital of Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Front Cardiovasc Med. 2023 Feb 8;10:1090938. doi: 10.3389/fcvm.2023.1090938. eCollection 2023.
Ocular abnormalities and the development of retinal vasculature may cause postnatal retinopathy. In the past decade, tremendous progress has been made in identifying the mechanisms that regulate retina vasculature. However, the means of regulating embryonic hyaloid vasculature development is largely unknown. This study aims to determine whether and how andrographolide regulates embryonic hyaloid vasculature development.
Murine embryonic retinas were used in this study. Whole mount isolectin B4 (IB4) staining, hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), and immunofluorescence staining (IF) were performed to determine whether andrographolide is critical for embryonic hyaloid vasculature development. BrdU incorporation assay, Boyden chamber migration assay, spheroid sprouting assay, and Matrigel-based tube formation assay were performed to evaluate whether andrographolide regulates the proliferation and migration of vascular endothelial cells. Molecular docking simulation and Co-immunoprecipitation assay were used to observe protein interaction.
Hypoxia conditions exist in murine embryonic retinas. Hypoxia induces HIF-1a expression; high-expressed HIF-1a interacts with VEGFR2, resulting in the activation of the VEGF signaling pathway. Andrographolide suppresses hypoxia-induced HIF-1a expression and, at least in part, interrupts the interaction between HIF-1a and VEGFR2, causing inhibiting endothelial proliferation and migration, eventually inhibiting embryonic hyaloid vasculature development.
Our data demonstrated that andrographolide plays a critical role in regulating embryonic hyaloid vasculature development.
眼部异常和视网膜血管系统的发育可能导致产后视网膜病变。在过去十年中，在确定调节视网膜血管系统的机制方面取得了巨大进展。然而，调节胚胎玻璃体血管系统发育的方式在很大程度上尚不清楚。本研究旨在确定穿心莲内酯是否以及如何调节胚胎玻璃体血管系统发育。
本研究使用小鼠胚胎视网膜。进行了全层异凝集素B4（IB4）染色、苏木精和伊红（H&E）染色、免疫组织化学（IHC）和免疫荧光染色（IF），以确定穿心莲内酯对胚胎玻璃体血管系统发育是否至关重要。进行了BrdU掺入试验、Boyden小室迁移试验、球体发芽试验和基于基质胶的管形成试验，以评估穿心莲内酯是否调节血管内皮细胞的增殖和迁移。使用分子对接模拟和免疫共沉淀试验观察蛋白质相互作用。
小鼠胚胎视网膜中存在缺氧情况。缺氧诱导HIF-1α表达；高表达的HIF-1α与VEGFR2相互作用，导致VEGF信号通路激活。穿心莲内酯抑制缺氧诱导的HIF-1α表达，并至少部分中断HIF-1α与VEGFR2之间的相互作用，从而抑制内皮细胞增殖和迁移，最终抑制胚胎玻璃体血管系统发育。
我们的数据表明，穿心莲内酯在调节胚胎玻璃体血管系统发育中起关键作用。
