慢病毒介导的多嘧啶束结合蛋白相关剪接因子抑制氧诱导视网膜病变小鼠视网膜新生血管

Q4 Medicine

中华眼底病杂志 Pub Date : 2020-01-25 DOI:10.3760/CMA.J.ISSN.1005-1015.2020.01.012

Liangyu Huang, Yifeng Ke, Tingting Lin, Shaochong Bu, X. Ren, Min Jiao, Yong Wang, Li-ying Hu, Qiong Wang, Y. Hong, Xiaorong Li

{"title":"慢病毒介导的多嘧啶束结合蛋白相关剪接因子抑制氧诱导视网膜病变小鼠视网膜新生血管","authors":"Liangyu Huang, Yifeng Ke, Tingting Lin, Shaochong Bu, X. Ren, Min Jiao, Yong Wang, Li-ying Hu, Qiong Wang, Y. Hong, Xiaorong Li","doi":"10.3760/CMA.J.ISSN.1005-1015.2020.01.012","DOIUrl":null,"url":null,"abstract":"Objective \nTo investigate the inhibitory effect of lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR). \n \n \nMethods \nOne hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group, simple OIR model group, OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group), with 16, 32, 32 and 32 mice, respectively. When the mice were 7 days old, the mice in the normal control group were fed in a routine environment, and the mice in the OIR model group, Vec group and PSF group were established OIR model. The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1×1011 TU/ml) at the age of 12 days. No injection was performed in the normal control group and simple OIR group. RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Western blot analysis was applied to detect the protein expression of Nrf2, HO-1 and PSF. \n \n \nResults \nOf the normal control group, simple OIR model group, Vec group and PSF group, the number of pre-retinal neovascular cell nuclei were 0.00, 14.36±5.50, 15.67±4.96, 8.13±2.09, the non-perfusion area were 0.00%, (35.71±2.81)%, (36.57±4.53)%, (15.33±4.75)%, respectively. The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87, 165.70; P<0.05). Compared with the normal control group, there were more pre-retinal neovascular cell nucleis and larger non-perfusion area in the simple OIR model group and Vec group (P<0.05). Compared with the simple OIR model group and Vec group, there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P<0.05). Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2, HO-1 (F=53.66, 83.54) and protein expression of Nrf2, HO-1 and PSF (F=58.38, 52.69, 24.79) among 4 groups were significant (P<0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P<0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P<0.05). model group and Vec group (P<0.05). \n \n \nConclusion \nIntravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1. \n \n \nKey words: \nRetinal neovascularization/preventionc Polypyrimidine traet-binding protein/drug effects; Lentivirus infections; NF-E2-related factor 2; Heme oxygenase (decyclizing); Animal experimentation","PeriodicalId":10103,"journal":{"name":"中华眼底病杂志","volume":"36 1","pages":"53-59"},"PeriodicalIF":0.0000,"publicationDate":"2020-01-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor inhibits retinal neovascularization in mice of oxygen-induced retinopathy\",\"authors\":\"Liangyu Huang, Yifeng Ke, Tingting Lin, Shaochong Bu, X. Ren, Min Jiao, Yong Wang, Li-ying Hu, Qiong Wang, Y. Hong, Xiaorong Li\",\"doi\":\"10.3760/CMA.J.ISSN.1005-1015.2020.01.012\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Objective \\nTo investigate the inhibitory effect of lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR). \\n \\n \\nMethods \\nOne hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group, simple OIR model group, OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group), with 16, 32, 32 and 32 mice, respectively. When the mice were 7 days old, the mice in the normal control group were fed in a routine environment, and the mice in the OIR model group, Vec group and PSF group were established OIR model. The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1×1011 TU/ml) at the age of 12 days. No injection was performed in the normal control group and simple OIR group. RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Western blot analysis was applied to detect the protein expression of Nrf2, HO-1 and PSF. \\n \\n \\nResults \\nOf the normal control group, simple OIR model group, Vec group and PSF group, the number of pre-retinal neovascular cell nuclei were 0.00, 14.36±5.50, 15.67±4.96, 8.13±2.09, the non-perfusion area were 0.00%, (35.71±2.81)%, (36.57±4.53)%, (15.33±4.75)%, respectively. The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87, 165.70; P<0.05). Compared with the normal control group, there were more pre-retinal neovascular cell nucleis and larger non-perfusion area in the simple OIR model group and Vec group (P<0.05). Compared with the simple OIR model group and Vec group, there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P<0.05). Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2, HO-1 (F=53.66, 83.54) and protein expression of Nrf2, HO-1 and PSF (F=58.38, 52.69, 24.79) among 4 groups were significant (P<0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P<0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P<0.05). model group and Vec group (P<0.05). \\n \\n \\nConclusion \\nIntravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1. \\n \\n \\nKey words: \\nRetinal neovascularization/preventionc Polypyrimidine traet-binding protein/drug effects; Lentivirus infections; NF-E2-related factor 2; Heme oxygenase (decyclizing); Animal experimentation\",\"PeriodicalId\":10103,\"journal\":{\"name\":\"中华眼底病杂志\",\"volume\":\"36 1\",\"pages\":\"53-59\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-01-25\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"中华眼底病杂志\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3760/CMA.J.ISSN.1005-1015.2020.01.012\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Medicine\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"中华眼底病杂志","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3760/CMA.J.ISSN.1005-1015.2020.01.012","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Medicine","Score":null,"Total":0}

引用次数: 1

摘要

目的探讨慢病毒介导的多嘧啶束结合蛋白相关剪接因子(PSF)对氧致视网膜病变(OIR)小鼠视网膜新生血管(RNV)的抑制作用。方法将12只5日龄C57BL/6J小鼠随机分为正常对照组、单纯OIR模型组、OIR模型+慢病毒空载体治疗组(Vec组)和OIR模型+ PSF慢病毒治疗组(PSF组)，分别设16只、32只、32只和32只。小鼠7日龄时，正常对照组小鼠在常规环境下饲养，OIR模型组、Vec组和PSF组小鼠分别建立OIR模型。Vec组和PSF组小鼠在12日龄时玻璃体内注射慢病毒载体和PSF慢病毒1 μl(滴度1×1011 TU/ml)。正常对照组和单纯OIR组均不注射。通过对小鼠视网膜进行免疫荧光染色，计数视网膜前新生血管细胞数和分析非灌注区来评估RNV。采用实时荧光定量PCR检测核因子-红细胞2相关因子- 2 (Nrf2)和血红素加氧酶-1 (HO-1) mRNA的表达。Western blot检测Nrf2、HO-1、PSF蛋白表达。结果正常对照组、单纯OIR模型组、Vec组、PSF组视网膜前新生细胞核数分别为0.00、14.36±5.50、15.67±4.96、8.13±2.09，非灌注面积分别为0.00%、(35.71±2.81)%、(36.57±4.53)%、(15.33±4.75)%。4组大鼠视网膜前新生细胞核数及非灌注面积比较差异均有统计学意义(F=24.87, 165.70;P < 0.05)。与正常对照组比较，单纯OIR模型组和Vec组视网膜前新生细胞核增多，非灌注面积增大(P<0.05)。与单纯OIR模型组和Vec组比较，PSF组视网膜前新生血管细胞核减少，非灌注面积减小(P<0.05)。实时荧光定量PCR和Western blot检测结果显示，4组大鼠Nrf2、HO-1 mRNA表达量(F=53.66、83.54)和Nrf2、HO-1、PSF蛋白表达量(F=58.38、52.69、24.79)差异均有统计学意义(P<0.05)。单纯OIR模型组和Vec组大鼠Nrf2、HO-1 mRNA表达量及Nrf2、HO-1、PSF蛋白表达量均显著低于正常对照组(P<0.05)。PSF组大鼠Nrf2、HO-1 mRNA表达量及Nrf2、HO-1、PSF蛋白表达量显著高于单纯OIR模型组和Vec组(P<0.05)。模型组和Vec组(P<0.05)。结论玻璃体内注射慢病毒介导的PSF可能通过上调Nrf2和HO-1的表达来抑制RNV。关键词:视网膜新生血管/预防多嘧啶治疗结合蛋白/药物效应;慢病毒感染;nf - e2相关因子2;血红素加氧酶(脱环);动物实验

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

Lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor inhibits retinal neovascularization in mice of oxygen-induced retinopathy

Objective To investigate the inhibitory effect of lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR). Methods One hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group, simple OIR model group, OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group), with 16, 32, 32 and 32 mice, respectively. When the mice were 7 days old, the mice in the normal control group were fed in a routine environment, and the mice in the OIR model group, Vec group and PSF group were established OIR model. The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1×1011 TU/ml) at the age of 12 days. No injection was performed in the normal control group and simple OIR group. RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Western blot analysis was applied to detect the protein expression of Nrf2, HO-1 and PSF. Results Of the normal control group, simple OIR model group, Vec group and PSF group, the number of pre-retinal neovascular cell nuclei were 0.00, 14.36±5.50, 15.67±4.96, 8.13±2.09, the non-perfusion area were 0.00%, (35.71±2.81)%, (36.57±4.53)%, (15.33±4.75)%, respectively. The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87, 165.70; P<0.05). Compared with the normal control group, there were more pre-retinal neovascular cell nucleis and larger non-perfusion area in the simple OIR model group and Vec group (P<0.05). Compared with the simple OIR model group and Vec group, there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P<0.05). Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2, HO-1 (F=53.66, 83.54) and protein expression of Nrf2, HO-1 and PSF (F=58.38, 52.69, 24.79) among 4 groups were significant (P<0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P<0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P<0.05). model group and Vec group (P<0.05). Conclusion Intravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1. Key words: Retinal neovascularization/preventionc Polypyrimidine traet-binding protein/drug effects; Lentivirus infections; NF-E2-related factor 2; Heme oxygenase (decyclizing); Animal experimentation

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

中华眼底病杂志 Medicine-Ophthalmology

CiteScore

0.40

自引率

0.00%

发文量

5383

期刊介绍： Chinese Journal of Ocular Fundus Diseases is the only scientific journal in my country that focuses on reporting fundus diseases. Its purpose is to combine clinical and basic research, and to give equal importance to improvement and popularization. It comprehensively reflects the leading clinical and basic research results of fundus disease disciplines in my country; cultivates professional talents in fundus disease, promotes the development of fundus disease disciplines in my country; and promotes academic exchanges on fundus disease at home and abroad. The coverage includes clinical and basic research results of posterior segment diseases such as retina, uveal tract, vitreous body, visual pathway, and internal eye diseases related to systemic diseases. The readers are medical workers and researchers related to clinical and basic research of fundus diseases. According to the journal retrieval report of the Chinese Institute of Scientific and Technological Information, the comprehensive ranking impact factor and total citation frequency of the Chinese Journal of Ocular Fundus Diseases have been among the best in the disciplines of ophthalmology, otolaryngology, and ophthalmology in my country for many years. The papers published have been included in many important databases at home and abroad, such as Scopus, Peking University Core, and China Science Citation Database (CSCD).