Yu-Han Yan, Hong-Yu Li, Li-Xiong Gao, Wen Li, Ling-Ping Zhao, Quan Zeng, Yu Luo, Tian-Tian Cui, Ru-Ge Zang, Zi Ye, Jia-Fei Xi, Wen Yue, Zhao-Hui Li
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引用次数: 0
Abstract
Aim: To explore the changes in early retinal development after the occurrence of ischemia.
Methods: Human retinal organoids (hROs) of day 18 or day 30 were treated with oxygen-glucose deprivation and reperfusion (OGD/R) to simulate the retinal ischemia. All hROs were maintained normally until day 60 to evaluate changes in ischemic injuries during retinal development. Paraffin section staining was used for detecting changes in organoid structure and cell number. Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB) analyses were used to observe the change in the expression of retinal cell markers.
Results: In hROs, OGD/R induced the decrease of proliferating cells, inhibited the expression of proliferated marker Ki67 and promoted early apoptosis of retinal cells (P<0.05). Under OGD/R condition, the progenitor cell layer and ganglion cell layer of hROs lost normal structure, and the number of neural stem cells (SOX2+), retinal progenitor cells (CHX10+) and retinal ganglion cells (TUJ1+/BRN3+/ATOH7+) decreased (P<0.05). The expression of corresponding retinal cell markers also decreased (P<0.05). Organoids treated with OGD/R on day 30 had similar injuries in retinal structure and retinal cell markers to those on day 18. Long-term observations revealed that day 18-treated organoids remained disorganized progenitor and ganglion cell layers by day 60, with no recovery in proliferating cell nuclear antigen (PCNA) protein expression. RT-qPCR showed persistently low Ki67 transcription levels (P<0.001), while other retinal cell markers recovered or exceeded normal levels, indicating a limited self-repair happened in the development of hROs. In contrast, day 30-treated organoids exhibited normal structure and marker expression by day 60, with transcription levels of retinal cell markers returning to normal (P>0.05), demonstrating complete recovery from OGD/R damage.
Conclusion: Retinal ischemia damage the retinal development in the short-term. After the restoration of retinal blood supply, the retinal ischemic damage can be recovered during subsequent development. However, retinal ischemic injuries at different developmental stages exhibit varying degrees of reversibility. The earlier ischemic injury occurs, the more difficult it is to repair retinal cell and structure damage.
目的：探讨缺血后视网膜早期发育的变化。方法：采用氧糖剥夺再灌注法（OGD/R）模拟视网膜缺血，对第18天和第30天的人视网膜类器官（hROs）进行处理。所有hROs维持正常至第60天，以评估视网膜发育过程中缺血性损伤的变化。石蜡切片染色检测类器官结构和细胞数量的变化。采用实时定量聚合酶链反应（RT-qPCR）和Western blot （WB）方法观察视网膜细胞标志物的表达变化。结果：在hROs中，OGD/R诱导增殖细胞减少，抑制增殖标志物Ki67的表达，促进视网膜细胞（P+）的早期凋亡，视网膜祖细胞（CHX10+）和视网膜神经节细胞（TUJ1+/BRN3+/ATOH7+）减少（PPPP>0.05），显示OGD/R损伤完全恢复。结论：视网膜缺血在短期内损害视网膜发育。视网膜血供恢复后，视网膜缺血性损伤可在后续发育中得到恢复。然而，不同发育阶段视网膜缺血性损伤表现出不同程度的可逆性。缺血性损伤发生越早，视网膜细胞和结构损伤的修复就越困难。
期刊介绍：
· International Journal of Ophthalmology-IJO (English edition) is a global ophthalmological scientific publication
and a peer-reviewed open access periodical (ISSN 2222-3959 print, ISSN 2227-4898 online).
This journal is sponsored by Chinese Medical Association Xi’an Branch and obtains guidance and support from
WHO and ICO (International Council of Ophthalmology). It has been indexed in SCIE, PubMed,
PubMed-Central, Chemical Abstracts, Scopus, EMBASE , and DOAJ. IJO JCR IF in 2017 is 1.166.
IJO was established in 2008, with editorial office in Xi’an, China. It is a monthly publication. General Scientific
Advisors include Prof. Hugh Taylor (President of ICO); Prof.Bruce Spivey (Immediate Past President of ICO);
Prof.Mark Tso (Ex-Vice President of ICO) and Prof.Daiming Fan (Academician and Vice President,
Chinese Academy of Engineering.
International Scientific Advisors include Prof. Serge Resnikoff (WHO Senior Speciatist for Prevention of
blindness), Prof. Chi-Chao Chan (National Eye Institute, USA) and Prof. Richard L Abbott (Ex-President of
AAO/PAAO) et al.
Honorary Editors-in-Chief: Prof. Li-Xin Xie(Academician of Chinese Academy of
Engineering/Honorary President of Chinese Ophthalmological Society); Prof. Dennis Lam (President of APAO) and
Prof. Xiao-Xin Li (Ex-President of Chinese Ophthalmological Society).
Chief Editor: Prof. Xiu-Wen Hu (President of IJO Press).
Editors-in-Chief: Prof. Yan-Nian Hui (Ex-Director, Eye Institute of Chinese PLA) and
Prof. George Chiou (Founding chief editor of Journal of Ocular Pharmacology & Therapeutics).
Associate Editors-in-Chief include:
Prof. Ning-Li Wang (President Elect of APAO);
Prof. Ke Yao (President of Chinese Ophthalmological Society) ;
Prof.William Smiddy (Bascom Palmer Eye instituteUSA) ;
Prof.Joel Schuman (President of Association of University Professors of Ophthalmology,USA);
Prof.Yizhi Liu (Vice President of Chinese Ophtlalmology Society);
Prof.Yu-Sheng Wang (Director of Eye Institute of Chinese PLA);
Prof.Ling-Yun Cheng (Director of Ocular Pharmacology, Shiley Eye Center, USA).
IJO accepts contributions in English from all over the world. It includes mainly original articles and review articles,
both basic and clinical papers.
Instruction is Welcome Contribution is Welcome Citation is Welcome
Cooperation organization
International Council of Ophthalmology(ICO), PubMed, PMC, American Academy of Ophthalmology, Asia-Pacific, Thomson Reuters, The Charlesworth Group, Crossref,Scopus,Publons, DOAJ etc.
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