[1]李玉平,王伶俐,陈震,等.circRNA_0051079通过靶向miR-26a-5p/PTEN轴诱导视网膜神经变性的分子机制[J].眼科新进展,2023,43(5):363-368.[doi:10.13389/j.cnki.rao.2023.0073]
　LI Yuping,WANG Lingli,CHEN Zhen,et al.Molecular mechanism of circular ribonucleic acid_0051079 inducing retinal neurodegeneration by targeted micro-ribonucleic acid-26a-5p/PTEN axis[J].Recent Advances in Ophthalmology,2023,43(5):363-368.[doi:10.13389/j.cnki.rao.2023.0073]
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circRNA_0051079通过靶向miR-26a-5p/PTEN轴诱导视网膜神经变性的分子机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年5期
页码:
363-368
栏目:
实验研究
出版日期:
2023-05-05

文章信息/Info

Title:
Molecular mechanism of circular ribonucleic acid_0051079 inducing retinal neurodegeneration by targeted micro-ribonucleic acid-26a-5p/PTEN axis
作者:
李玉平; 王伶俐; 陈震; 宋辉; 姚为华; 刘易
438000　湖北省黄冈市，黄冈市中心医院眼科（李玉平，宋辉，姚为华，刘易）；438300　湖北省黄冈市，麻城市人民医院眼科（王伶俐)；438000　湖北省武汉市，湖北省人民医院眼科（陈震）
Author(s):
LI Yuping1; WANG Lingli2; CHEN Zhen3; SONG Hui1; YAO Weihua1; LIU Yi1
1.Department of Ophthalmology，Huanggang Central Hospital，Huanggang 438000,Hubei Province,China
2.Department of Ophthalmology，Macheng People’s Hospital，Huanggang 438300，Hubei Province,China
3.Department of Ophthalmology，Hubei Provincial People’s Hospital，Wuhan 438000，Hubei Province,China
关键词:
视网膜缺血; 缺血/再灌注; 环状RNA; miR-26a-5p/PTEN通路; 视网膜神经变性
Keywords:
retinal ischemia;  ischemia/reperfusion;  circular ribonucleic acid;  micro-ribonucleic acid-26a-5p/PTEN pathway;  retinal neurodegeneration
分类号:
R774.1
DOI:
10.13389/j.cnki.rao.2023.0073
文献标志码:
A
摘要:
目的　探讨circ_0051079对缺血/再灌注（I/R）损伤诱导的视网膜神经变性的影响及机制。
方法　从C57BL/6J乳鼠眼球中分离视网膜神经节细胞（RGC），随机分为对照组、siRNA组（转染阴性siRNA）、si_circ_0051079组（转染si-circ-0051079干扰RNA）、模拟物对照组（转染Scr mimic）、miR-26a-5p组（转染miR-26a-5p mimic）、miR-26a-5p+vector组（转染pcDNA 3.1）、miR-26a-5p+PTEN组（转染pcDNA 3.1-PTEN），分别在正常、缺氧（体积分数1%O2暴露）或氧化应激（50 μmol·L-1H2O2暴露）条件下培养24 h，进行RT-qPCR、CCK-8、TUNEL、RNA免疫沉淀（RIP）等检测。于15只C57BL/6小鼠左眼中建立I/R损伤模型，对侧眼保持正常眼压作为对照，I/R损伤后0 d、3 d和7 d各取5只小鼠收集视网膜检测circ_0051079表达。另取20只C57BL/6小鼠分为正常对照组（用针头插入左眼前房但不升高眼压）、I/R 组（左眼I/R损伤处理）、I/R+对照shRNA组（左眼注射无序shRNA并建立I/R损伤模型）、I/R+circ_0051079 shRNA组（左眼注射circ_0051079 shRNA并建立I/R损伤模型）。I/R损伤后7 d，取视网膜进行蛋白免疫荧光染色。收集2020年11月至2021年3月急性闭角型青光眼和白内障患者的房水样本各20例，进行RT-qPCR测定。
结果　缺氧或氧化应激条件下培养的RGC中circ_0051079表达量较正常条件下培养的RGC增加（均为P<0.05）。在缺氧或氧化应激条件下培养，si_circ_0051079组RGC细胞活力均较siRNA组增加，RGC凋亡细胞数均减少（均为P<0.05）。荧光素酶报告基因分析和RIP检测结果表明，circ_0051079可以通过调节miR-26a-3p/PTEN信号转导进而影响RGC功能。动物实验中，I/R损伤可致视网膜组织中circ_0051079表达量升高（P<0.05）。与正常对照组（1.00±0.07）相比，I/R+circ_0051079 shRNA组小鼠视网膜中circ_0051079表达水平（0.37±0.04）下降（P<0.05），同时I/R诱导的视网膜神经变性减轻。临床研究中，与白内障患者房水样本相比，青光眼患者房水样本中circ_0051079和PTEN mRNA表达增加，miR-26a-3p表达降低（均为P<0.001）。
结论　circ_0051079可能是缺血性视网膜病变诊断和治疗的潜在靶点。
Abstract:
Objective　To investigate the effect and mechanism of circ_0051079 on retinal neurodegeneration induced by ischemia/reperfusion (I/R).
Methods　Retinal ganglion cells (RGCs) were isolated from the eyeballs of C57BL/6J suckling mice and divided into the control group, siRNA group (transfected with negative siRNA), si_circ_0051079 group (transfected with si_circ_0051079 interfering RNA), mimic control group (transfected with Scr mimic), miR-26a-5p group (transfected with miR-26a-5p mimic), miR-26a-5p+vector group (transfected with pcDNA 3.1), and miR-26a-5p+PTEN group (transfected with pcDNA 3.1-PTEN). RGCs were cultured under the normal, hypoxia (1% O2 exposure) and oxidative stress (50 μmol·L-1 H2O2 exposure) conditions for 24 h, respectively, and then detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Cell Counting Kit-8, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and RNA immunoprecipitation (RIP). I/R injury model was established in the left eye of 15 C57BL/6 mice, and normal intraocular pressure was maintained in the contralateral eye as a control. Five mice were selected to collect retinas at 0 d, 3 d and 7 d after I/R injury modeling to detect the expression of circ_0051079. Another 20 C57BL/6 mice were selected and divided into the control group (left eye was injected with a needle without increasing intraocular pressure), I/R group (I/R injury model was established), I/R+control shRNA group (left eye was injected with disorder shRNA and I/R injury model was established), I/R+ circ_0051079 shRNA group (left eye was injected with circ_0051079 shRNA and I/R injury model was established). Retinal tissues were taken 7 d after I/R injury, and detected by protein immunofluorescence staining. Aqueous humor (AH) samples from 20 patients with acute angle-closure glaucoma and 20 patients with cataract in our hospital from November 2020 to March 2021 were collected and determined by RT-qPCR.
Results　The expression level of circ_0051079 in RGCs cultured under hypoxia or oxidative stress was significantly higher than in normal conditions (both P<0.05). Under hypoxia or oxidative stress, the activity of RGCs in the si_circ_0051079 group increased compared with the siRNA group, and the number of apoptotic RGCs decreased (both P<0.05). Luciferase reporter assay and RIP showed that circ_0051079 could affect the functions of RGCs by regulating miR-26a-3p/PTEN signal transduction. The animal experiments showed that the expression level of circ_0051079 in retinal tissue significantly increased after I/R injury (P<0.05). Compared with the control group (1.00±0.07), the expression of circ_0051079 in retinal tissue significantly decreased in the I/R+circ_0051079 shRNA group (0.37±0.04) (P<0.05), and I/R-induced retinal neurodegeneration was relieved. In clinical studies, compared with the AH samples of cataract patients, the expressions of circ_0051079 and PTEN significantly increased, and the expression of miR-26a-3p significantly decreased in the AH of glaucoma patients(all P<0.001) .
Conclusion　The circ_0051079 may be a promising target for the diagnosis and treatment of ischemic retinopathy.

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更新日期/Last Update: 2023-05-05