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Efficacy of intravitreal captopril on oxygen-induced retinopathy in mice

International Journal of Ophthalmology(English edition) 2011年第4期4卷 361-364页

作者： Yu Di, Xiao-Long Chen Department of Ophthalmology Shengjing Hospital of China Medical University Shenyang 110004 Liaoning Province China Graduate School of China Medical University Shenyang 110001 Liaoning Province China

AIM: To study the inhibitory effect of intravitreal captopril on oxygen-induced retinopathy (OIR) in mice. METHODS: Eighty postnatal day (P)7 C57BL/63 mice were randomly divided into treated group and control group with forty mice in each group. The mice were exposed to 75% 2% oxygen for 5 days (P7-P11) and then returned to room air for 5 days (P12-P17) to induce retinal neovascularization (RNV). Beginning on P12, the mice in treated group received daily intravitreal injections of captopril(3.0mL/kg), while those in control group received daily intravitreal injections of phosphate-buffered saline (PBS) (3.0mL/kg) through P17. After anesthetized at P17, one eye was chosen randomly as experimental eye and were enucleated. RNV was examined by Adenosine diphosphate-ase (ADPase) stained retina flat-mounts and was quantitated histologically by counting the neovascular endothelial cell nuclei anterior to inner limiting membrane (ILM). The expressions of matrix metalloproteinase-2 (MMP-2) and vascular endothelial growth factor (VEGF) were measured by immunohistochemical method. RESULTS: Comparing with control group, more regular distributions, better branch and reduced density of RNV were observed in eyes of treated group. The number of neovascular cell nuclei was less in treated group than that in control group ( t =6.135, P <0.01). Stain of MMP-2 and VEGF was weaker in treated group than that in control group. CONCLUSION: The results indicate that captopril can significantly inhibit RNV in OIR mice.

关键词： retinal neovascularization oxygen-induced retinopathy captopril intravitreal injection

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Acetylcholinesterase inhibition ameliorates retinal neovascularization and glial activation in oxygen-induced retinopathy

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International Journal of Ophthalmology(English edition) 2020年第9期13卷 1361-1367页

作者： Qiu-Ping Liu Xian Zhang Ya-Zhou Qin Jing-Lin Yi Jing-Ming Li Affiliated Eye Hospital of Nanchang University Nanchang 330006Jiangxi ProvinceChina Department of Ophthalmology First Affiliated Hospital of Xi'an Jiaotong UniversityXi’an 710061Shaanxi ProvinceChina

AIM:To investigate whether inhibition of acetylcholinesterase(AChE)by donepezil ameliorate aberrant retinal neovascularization(RNV)and abnormal glial activation in oxygen-induced retinopathy(OIR).METHODS:A mouse model of RNV was induced in postnatal day 7(P7)mice by exposure to 75%oxygen.Donepezil was administrated to P12 mice by intraperitoneal injection.Expression and localization of AChE in mouse retinas were determined by immunofluorescence.RNV was evaluated by paraffin sectioning and hematoxylin and eosin(HE)staining.Activation of retinal M uller glial cells were examined by immunoblot of glial fibrillary acidic protein(GFAP).rMC-1.a retinal Muller cell line,was used for in vitro study.Expression of hypoxia-induced factor 1α(HIF-1α)and vascular endothelial growth factor(VEGF)were determined by Western-blot analysis,enzyme-linked immunosorbent assay(ELISA)or immunostaining.RESULTS:Aberrant RNV and glial activation was observed after OIR.Of note,retinal AChE was mainly expressed by retinal Muller glial cells and markedly increased in OIR mice.Systemic administration of donepezil significantly reduced RNV and abnormal glial activation in mice with OIR.Moreover,ischemia-induced HIF-1αaccumulation and VEGF upregulation in OIR mouse retinas and cultured rMC-1 were significantly inhibited by donepezil intervention.CONCLUSION:AchE is implicated in RNV with OIR.Inhibition of AChE by donepeizl is likely to be a potential therapeutic approach for retinal neovascular diseases.

关键词： acetylcholinesterase oxygen-induced retinopathy neovascularization glosis mice

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Activated complement classical pathway in a murine model of oxygen-induced retinopathy

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International Journal of Ophthalmology(English edition) 2015年第1期8卷 17-22页

作者： Xue-Ying Tao Shi-Jie Zheng Bo Lei Department of Ophthalmology the First Affiliated Hospital of Chongqing Medical UniversityChongqing Key Laboratory of OphthalmologyChongqing Eye Institute Department of Ophthalmology Chongqing Maternal and Child Health-Care Hospital

AIM: To investigate whether the complement system is involved in a murine model of oxygen-induced retinopathy(OIR).METHODS: Forty C57BL/6J newborn mice were divided randomly into OIR group and control group. OIR was induced by exposing mice to 75% ±2% oxygen from postnatal 7d(P7) to P12 and then recovered in room air.For the control group, the litters were raised in room air.At the postnatal 17d(P17), gene expressions of the complement components of the classical pathway(CP),the mannose-binding lectin(MBL) pathway and the alternative pathway(AP) in the retina were determined by quantitative real-time polymerase chain reaction(RT-PCR). Retinal protein expressions of the key components in the CP were examined by Western blotting.· RESULTS: Whole mounted retina in the OIR mice showed area of central hypoperfusion in both superficial and deep layers and neovascular tufts in the periphery.The expressions of C1 qb and C4 b genes in the OIR retina were significantly higher than those of the controls. The expression of retinal complement factor B(CFB) gene in OIR mice was significantly lower than those of the controls. However, the expressions of C3 and complement factor H(CFH) genes were higher. The protein synthesis of the key components involved in the CP(C1q, C4 and C3) were also significantly higher in OIR mouse retina. Although MBL-associated serine protease 1(MASP1) and MASP2 were detected in both the OIR and the control groups, the expressions were weak and the difference between the two groups was not significant.CONCLUSION: Our data suggest that the complement system CP is activated during the pathogenesis of murine model of OIR.

关键词： oxygen-induced retinopathy complement activation classical pathway retina mouse

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Role of unc5b in retinal neovascularization in mice with oxygen-induced retinopathy

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International Journal of Ophthalmology(English edition) 2011年第2期4卷 137-142页

作者： Dan Liu, Lei Shang Department of Ophthalmology Xiangya Hospital of the Central South University Changsha 410013 Hunan Province China Department of Anatomy and Neurobiology Xiangya Medical School the Central South University Changsha 410013 Hunan Province China

AIM: To explore the role of unc5b in retinal neovascularization in murine oxygen-induced retinopathy (OIR). METHODS: On postnatal 7 (P7), C57BL/6J mice were exposed to 75% +/- 2% oxygen for 5 days. On postnatal 12 (P12), the mice were brought back to the room air (21% oxygen) to induce retinal neovascularization. Western blot analysis was performed to examine the temporal expression of unc5b in murine retinas. Double staining for unc5b and isolectin B4 were employed to determine the location of unc5b in murine retinas. The effect of unc5b on retinal neovascularization was evaluated by intravitreal injection of unc5b-FC in mice with OIR. Retinal neovascularization was measured by counting neovascular cell nuclei above the internal limiting membrane and by angiography of flat-mounted retinas perfused with fluorescein dextran. o RESULTS: Compared to age-matched normal mice, the expression of unc5b was significantly increased in retinas of OIR mice on P17 and P21. Unc5b was apparently expressed in retinal vessels of OIR while being negative in normal retinal vessels. Retinal neovascularization in eyes injected with unc5b-FC was significantly reduced. CONCLUSION: Unc5b-FC can effectively inhibit retinal neovascularization induced by OIR. It may serve as a powerful and novel therapy for ischemia-induced retinal disease.

关键词： oxygen-induced retinopathy unc5b unc5b-FC

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Expression of cyclinD1 in a rat model of oxygen-induced retinopathy

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Neural Regeneration Research 2011年第21期6卷 1651-1655页

作者： You Li Yuqiang Zhang Shaoyang Shi Xiaolong Chen Department of Ophthalmology Shengjing Hospital China Medical University Shenyang 110004 Liaoning Province China Department of Orthopedics First Affiliated Hospital China Medical University Shenyang 110001 Liaoning Province China

This study sought to elucidate the correlation between cyclinD1 expression and the emergence of neovascularization in oxygen-induced retinopathy （OIR）. OIR was induced in Sprague-Dawley 7-day-old neonatal rats exposed to hyperoxia （75% O2） for 5 days, and then returned to room air. Adenosine diphosphatase staining showed that the neovascularization started to emerge at rat age of day 14, and reached a peak at day 17, then gradually decreased and resolved by day 26. Immunohistochemical and immunofluorescence staining revealed that cyclinD1 protein expression was seen in the OIR rats at the age of day 12, then gradually increased and returned to normal levels by day 26. These experimental findings demonstrated that the temporal pattern of cyclinD1 protein expression is consistent with the emergence of retinal neovascularization.

关键词： oxygen-induced retinopathy cyclinD1 neovascularization rat

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Maternally expressed gene 3 regulates retinal neovascularization in retinopathy of prematurity

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Neural Regeneration Research 2022年第6期17卷 1364-1368页

作者： Yu Di Yue Wang Yue-Xia Wang Xue Wang Yuan Ma Qing-Zhu Nie Department of Ophthalmology Shengjing Hospital of China Medical UniversityShenyangLiaoning ProvinceChina

The mouse model of oxygen induced retinopathy is suitable for the study of various retinal neovascularization diseases,including retinopathy of prematurity.The maternally expressed gene 3(MEG3)has been demonstrated to have an inhibitory effect on diabetic retinopathy.In this study,we investigated the role of MEG3 overexpression in oxygen-induced retinopathy in mice.The results showed that MEG3 overexpression effectively inhibited the production of retinal neovascularization in oxygen-induced retinopathy mice.It acts by down-regulating the expression of phosphoinositide 3-kinase,serine/threonine kinase,and vascular endothelial growth factor and pro-inflammatory factors.MEG3 overexpression lentivirus has a future as a new method for the clinical treatment of retinopathy of prematurity.The animal experiments were approved by the Animal Ethics Committee of Shengjing Hospital of China Medical University,China(approval No.2016PS074K)on February 25,2016.

关键词： long noncoding RNA maternally expressed gene 3 neurodevelopment oxygen-induced retinopathy phos

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microRNA-218 Inhibits oxygen-induced Retinal Neovascularization via Reducing the Expression of Roundabout I

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Chinese Medical Journal 2016年第6期129卷 709-715页

作者： Shuang Han Yi-Chun Kong Bei Sun Quan-Hong Han Ying Chen Yu-ChuanWang Clinical College of Ophthalmology Tianjin Medical University Tianjin 300020 China Department of General Ophthalmology Tianjin Eye Hospital Tianjin Eye Institute Tianjin Key Laboratory of Ophthalmology and Visual Science Tianjin 300020 China Key Laboratory of Hormones and Development Ministry of Health Metabolic Diseases Hospital and Tianjin Institute of Endocrinology Tianjin Medical University Tianjin 300020 China Department of Vitreous and Retina Diseases Tianjin Eye Hospital Tianjin Eye Institute Tianjin Key Laboratory of Ophthalmology and Visual Science Tianiin 300020 China Department of Pathology Tianjin Eye Hospital Tianjin Eye Institute l:ianjin Key Laboratory of Ophthalmology and Visual Science Tianjin 300020 China

Background： The mechanisms of pathological retinal neovascularization （RNV） remain unknown. Several microRNAs were reported to be involved in the process of RNV. oxygen-induced retinopathy （O1R） is a useful model to investigate RNV. Our present work explored the expression and the role of microRNA-128 （miR-218） in oxygen-induced RNV. Methods： OIR was used to establish RNV model. The expression level ofmiR-218 in the retina from OIR mice was assessed by quantitative real-time reverse transcriptase polymerase chain reaction. Fluorescein angiography was performed in retinae of OIR mice, and RNV was quantified by hematoxylin and eosin staining to evaluate the effect of pCDH-CMV-miR-218 intravitreal injection on RNV in OIR mice. Roundabout 1 （Robol） expression was detected by Western blotting in mouse retinal vascular endothelial cells expressing a high or low level of miR-218 and retinal tissues from OIR mice. Cell migration was evaluated by scratch wound assay. Results： In OIR mice, the expression level of miR-218 was significantly down-regulated （P = 0.006）. Retinal Robol expression was significantly increased at both mRNA and protein levels （P = 0.001, 0.008： respectively）, miR-218 intravitreal injection inhibited retinal angiogenesis in OIR mice, and the restoration of miR-218 in retina led to down-regulation of Robol. Conelusions： Our experiments showed that restoration ofmiR-218 inhibited retinal angiogenesis via targeting Robo 1. MiR-218 contributed to the inhibition of retinal angiogenesis and miR-218 might be a new therapeutic target for preventing RNV.

关键词： miR-218 oxygen-induced retinopathy Retinal Neovascularization Roundabout 1

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Expression and function of Delta-like ligand 4 in a rat model of retinopathy of prematurity

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Neural Regeneration Research 2013年第8期8卷 723-730页

作者： Shaoyang Shi Xun Li You Li Cunwen Pei Hongwei Yang Xiaolong Chen Department of Ophthalmology Shengjing HospitalChina Medical University Department of Ophthalmology the 230 Hospital of Chinese PLA

The Delta-like ligand 4/Notch signaling pathway was shown to participate in the process of retinal development and angiogenesis. However, the function of the Delta-like ligand 4/Notch signaling pathway in retinopathy of prematurity requires further study. retinopathy of prematurity was induced in 5-day-old Sprague-Dawley rats exposed to hyperoxia for 7 days, and then returned to room air. Reverse transcription-PCR and western blot revealed that Delta-like ligand 4 levels decreased at postnatal day 12 and increased at postnatal day 17 in retinopathy of prematurity rats. Flat-mounted adenosine diphosphatase stained retina and hematoxylin-eosin stained retinal tissue slices showed that the clock hour scores and the nuclei counts in retinopathy of prematurity rats were significantly different compared to normal control rats. After retinopathy of prematurity rats were intravitreally injected with Delta-like ligand 4 monoclonal antibody to inhibit the Delta-like ligand 4/Notch signaling pathway, there was a significant increase in the severity of retinal neovascularization （clock hours） in the intravitreally injected eyes. The nuclei count was highly correlated with the clock hour score. These results suggest that Delta-like ligand 4/Notch signaling plays an essential role in the process of physiological and pathological angiogenesis in the retina.

关键词： neural regeneration peripheral nerve injury Delta-like ligand 4 retinopathy of prematurity retinalneovascularization vascular endothelial cells vascular endothelial growth factor Notch signalingpathway oxygen-induced retinopathy optic nerve disease photographs-containing paper neuroregeneration

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The progress of prophylactic treatment in retinopathy of prematurity

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International Journal of Ophthalmology(English edition) 2018年第5期11卷 858-873页

作者： Hong-Bing Zhang Xiao-Dong Wang Kun Xu Xiao-Gang Li Eye Institute of Shaanxi Province Xi'an First Hospital Xi'an 710002 Shaanxi Province China Department of Internal Medicine Department of Biochemistry and Molecular Biology University of Kansas Medical CenterKansas City KS 66160 USA

retinopathy of prematurity（ROP） is a retinal vascular disorder frequently found in premature infants.Different therapeutic strategies have been developed to treat ROP.However,there are still many children with ROP suffering by severe limitations in vision or even blindness.Recently,ROP has been suggested to be caused by abnormal development of the retinal vasculature,but not simply resulted by retinal neovascularization which takes about 4 to 6 wk after birth in premature infants.Thus,instead of focusing on how to reduce retinal neovascularization,understanding the pathological changes and mechanisms that occur prior to retinal neovascularization is meaningful,which may lead to identify novel target（s） for the development of novel strategy to promote the healthy growth of retinal blood vessels rather than passively waiting for the appearance of retinal neovascularization and removing it by force.In this review,we discussed recent studies about,1） the pathogenesis prior to retinal neovascularization in oxygen-induced retinopathy（OIR;a ROP in animal model） and in premature infants with ROP;2） the preclinical and clinical research on preventive treatment of early OIR and ROP.We will not only highlight the importance of the mechanisms and signalling pathways in regulating early stage of ROP but also will provide guidance for actively exploring novel mechanisms and discovering novel treatments for early phase OIR and ROP prior to retinal neovascularization in the future.

关键词： retinopathy of prematurity oxygen-induced retinopathy retinal neovascularization

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Placental growth factor expression is reversed by anti-vascular endothelial growth factor therapy under hypoxic conditions

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World Journal of Pediatrics 2014年第3期10卷 262-270页

作者： Ai-Yi Zhou Yu-Jing Bai Min Zhao Wen-Zhen Yu Lv-Zhen Huang Xiao-Xin Li Department of Ophthalmology the Second Affi liated Hospital of Medical College of Xi'an Jiaotong UniversityXi'an 710004Shaanxi ProvinceChina Key Laboratory of Vision Loss and Restoration Ministry of EducationDepartment of OphthalmologyPeople's HospitalPeking UniversityBeijingChina

Background:Clinical trials have revealed that the antivascular endothelial growth factor(VEGF)therapies are effective in retinopathy of prematurity(ROP).But the low level of VEGF was necessary as a survival signal in healthy conditions,and endogenous placental growth factor(PIGF)is redundant for development.The purpose of this study was to elucidate the PIGF expression under hypoxia as well as the infl uence of anti-VEGF therapy on PIGF.Methods:CoCl2-induced hypoxic human umbilical vein endothelial cells(HUVECs)were used for an in vitro study,and oxygen-induced retinopathy(OIR)mice models were used for an in vivo study.The expression patterns of PIGF under hypoxic conditions and the infl uence of anti-VEGF therapy on PIGF were evaluated by quantitative reverse transcription-polymerase chain reaction(RTPCR).The retinal avascular areas and neovascularization(NV)areas of anti-VEGF,anti-PIGF and combination treatments were calculated.Retina PIGF concentration was evaluated by ELISA after treatment.The vasoactive effects of exogenous PIGF on HUVECs were investigated by proliferation and migration studies.Results:PIGF mRNA expression was reduced by hypoxia in OIR mice,in HUVECs under hypoxia and anti-VEGF treatment.However,PIGF expression was reversed by anti-VEGF therapy in the OIR model and in HUVECs under hypoxia.Exogenous PIGF significantly inhibited HUVECs proliferation and migration under normal conditions,but it stimulated cell proliferation and migration under hypoxia.Anti-PIGF treatment was effective for neovascular tufts in OIR mice(P<0.05).Conclusion:The finding that PIGF expression is iatrogenically up-regulated by anti-VEGF therapy provides a consideration to combine it with anti-PIGF therapy.

关键词： human umbilical vein endothelial cells oxygen-induced retinopathy placental growth factor vascular endothelial growth factor

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