论著

2型糖尿病视网膜病变严重程度与血清生化全套指标的相关性

Correlation between the severity of type 2 diabetic retinopathy and the serum biochemical indexes

:200-204
 
目的:分析不同程度2型糖尿病视网膜病变的血清生化全套指标,探讨各生化指标与糖尿病性视网膜病变(diabetic retinopathy,DR)程度的相关性。方法:回顾性收集2018年5月至2020年10月在福建医科大学附属第二医院就诊的2型糖尿病患者。根据眼底情况分为3组:糖尿病不伴视网膜病变(non-diabetic retinopathy,NDR)组(17例)、糖尿病伴非增殖性视网膜病变(non-proliferative diabetic retinopathy,NPDR)组(29例)、糖尿病伴增殖期视网膜病变(proliferative diabetic retinopathy,PDR)组(34例)。采用SPSS 24.0分析比较3组血清生化全套各指标水平,对差异指标与DR相关性采用Pearson相关分析与多重线性回归分析。结果:3组年龄差异无统计学意义(P>0.05);3组的总蛋白、白蛋白、白球比、钙浓度差异均有统计学意义(均P<0.05)。组间比较发现NPDR和PDR组总蛋白、白蛋白、白球比、钙浓度低于NDR组,差异有统计学意义(P<0.05)。Pearson相关分析显示总蛋白(r=?0.290)、白蛋白(r=?0.304),钙浓度(r=?0.252)与DR呈负相关(均P<0.05),多重线性回归提示总蛋白、白蛋白及血钙浓度是DR保护因素(P<0.05)。结论:血清指标总蛋白、白蛋白及血钙浓度与DR成负相关,在生理范围内将这些指标维持相对较高的水平,或许可以为糖尿病视网膜病变的防治提供新思路。
Objective: To analyze the serum biochemical indexes of different degrees of type 2 diabetic retinopathy, and the correlation between biochemical indexes and the degree of diabetic retinopathy (DR). Methods: Patients with type 2 diabetes admitted to the Fujian Medical University 2nd Affiliate Hospital from May 2018 to October 2020 were retrospectively collected. The patients were divided into 3 groups according to fundus conditions: non-diabetic retinopathy (NDR) group (n=17), diabetes with non-proliferative diabetic retinopathy (NPDR) group (n=29),diabetes with proliferative diabetic retinopathy (PDR) group (n=34). SPSS 24.0 was used to compare the levels of a complete set of serum biochemical indexes in the three groups. Pearson correlation analysis and multiple linear regression analysis were used for the correlation between the different indexes and DR. Results: There was no statistically significant difference in age between the three groups. The differences in total protein, albumin, white sphere ratio, and calcium concentration of the three groups were statistically significant (P<0.05). The comparison between the groups showed that the total protein, albumin, white sphere ratio, and calcium concentration in the NPDR and PDR groups were lower than those in the NDR group. The difference was statistically significant (P<0.05). Pearson correlation analysis showed that total protein (r=?0.290), albumin (r=?0.304), and
calcium concentration (r=?0.252) were negatively correlated with DR (all P<0.05). Multiple linear regression indicated total protein, albumin and blood calcium concentration were the protective factors of DR (P<0.05).Conclusion: This study found that serum levels of total protein, albumin, and blood calcium levels are negatively correlated with DR. Maintaining a relatively high level of these indexes within the physiological range may provide new ideas for the prevention and treatment of diabetic retinopathy.
论著

玻璃膜疣主要成分胆固醇对人视网膜色素上皮细胞金属硫蛋白表达的影响

Effect of cholesterol exposure on the expression of metallothionein in human retinal pigment epithelial cells

:128-132
 
目的:研究玻璃膜疣主要成分胆固醇对人视网膜色素上皮细胞ARPE-19中金属硫蛋白表达的影响。方法:体外培养ARPE-19细胞,将细胞分为对照组和胆固醇处理组(2.5 mg/mL),取样时间为0,6,12,24,48,72h。通过实时定量PCR检测hMT1a,hMT2a和hMT3在转录水平的表达,用蛋白质印迹法检测总金属硫蛋白的表达。结果:在转录水平上hMT1a,hMT2a和hMT3受到胆固醇影响mRNA表达上调,且hMT3上调倍数最大;总金属硫蛋白的蛋白表达随着胆固醇处理时间延长明显增多。结论:玻璃膜疣主要成分胆固醇可以上调人视网膜色素上皮细胞中金属硫蛋白的表达,提示金属硫蛋白表达可受到玻璃膜疣形成起始阶段的刺激,其检测是否能用于年龄相关性黄斑变性的早期发现及早期诊断还需深入探讨。
Objective: To study the effects of cholesterol, the main component of drusen, on the expression of metallothionein of ARPE-19 cells. Methods: The ARPE-19 cell line was cultured in vitro, and the cells were divided into a control group and a cholesterol treatment group (2.5 mg/mL). The treatment time was 0, 6, 12, 24, 48, 72 hours. Real- time quantitative PCR was used to detect the expression of hMT1a, hMT2a and hMT3 at the mRNA level, and Western blot was used to detect the expression at the protein level. Results: The mRNA expression of hMT1a, hMT2a and hMT3 were up-regulated by cholesterol and the protein expression of total MTs was increased with cholesterol treatment. Conclusion: Cholesterol, the main component of drusen, can up-regulate the expression of metallothionein in human retinal pigment epithelial cells, suggesting that the expression of metallothionine can be stimulated by the initial stage of drusen formation. However, whether its detection can be used for the early detection and early diagnosis of age-related macular degeneration or not still needs to be further explored.
眼底内专栏

结核性脉络膜结节联合视网膜结节1例

A case report of choroidal tubercle complicated with intraretinal tubercle

:122-126
 
眼结核的表现形式多样,有时会被误诊漏诊。本病例为一年轻男性,以右眼视力突发下降为主诉就诊,眼部表现可见黄斑区脉络膜占位病灶及视网膜内小病灶,经γ-干扰素释放试验、胸部CT等检查,确诊为血型播散性肺结核,眼部诊断为结核性脉络膜结节联合视网膜结节,全身接受抗结核治疗后,眼部病灶消退。
Ocular tuberculosis encompasses a variety of clinical manifestations, which is easily misdiagnosed sometimes. In this article, a young male was admitted to our hospital with a chief complaint of sudden blurred vision in his right eye. His ocular examination detected a tiny mass in the choroid and a small lesion in the retina. After interferon-gamma release assay (IGRA) and chest CT scan, the patient was diagnosed with military tuberculosis. The ocular examination confirmed the diagnosis of choroidal tubercle complicated with intrarentinal tubercle, which were healed after systemic anti-tuberculosis therapy.
眼底内专栏

光学相干断层扫描成像对急性中心性浆液性脉络膜视网膜病变渗漏点的前瞻性观察研究

Observation on the leakage point of acute central serous chorioretinopathy through optical coherence tomography: A prospective study

:115-121
 
目的:观察急性中心性浆液性脉络膜视网膜病变(central serous chorioretinopathy,CSC)的自然病程中渗漏点的形态及变化。方法:本研究为前瞻性研究,使用光学相干断层扫描(optical coherence tomography,OCT)观察从发病到发病后5~13个月的CSC患者的渗漏点的OCT形态,测量并计算Hall层、脉络膜全层各自厚度及比值,并进行比较。结果:共20例患者[男14例,女6例,年龄33~59(中位数41)岁]纳入研究。随访时间为5~13个月。在19例患者中观察到微小视网膜色素上皮脱离(pigment epithelium detachment,PED)。1例患者可见视网膜色素上皮(retinal pigment epithelium, RPE)小凸起。在随访期间,仅1例患者的PED完全恢复,其他19例患者在视网膜下液被完全吸收时,RPE和Bruch膜之间仍存在微小分离。渗漏点处的Haller层/脉络膜厚度显著高于中央凹处(初诊时0.806±0.08 vs 0.863±0.06,P=0.003;最后1次随访时为0.801±0.07 vs 0.851±0.06,P=0.004)。结论:本研究观察到在急性CSC患者自然病程中,即使视网膜下液吸收,OCT显示渗漏点处仍存在持续的PED,更厚的Haller层及更薄的内层脉络膜,这些发现为CSC的发病机制提供了更多线索。
Objective: To observe the morphology and changes of leakage points in the natural course of acute central serous chorioretinopathy (CSC). Methods: This study was a prospective study, using optical coherence tomography to observe the OCT morphology of leakage points in CSC patients from onset to 5 to 13 months after onset, measuring the thickness and ratio of Hall layer and the whole choroid, and then compare them. Results: A total of 20 patients were included in the study, including 14 males and 6 females, aged from 33 to 59, with the median being 41 years old. The follow-up time ranged from 5 months to 13 months. Minute retinal pigment epithelial detachments (PED) were observed in 19 patients. A small bulge of retinal pigment epithelium (RPE) was observed in 1 patient. During the follow-up, only one patient totally recovered. Small separation between RPE and Bruch membrane still exit even subretinal fluid were absorbed completely in the other 19 patients. The thickness of Haller layer or choroid at the leakage point was significantly higher than that of the fovea (0.806±0.08 vs 0.863±0.06, P=0.003, at the first visit; 0.801±0.07 vs 0.851±0.06, P=0.004, at the last follow-up). Conclusion: This study observed that in the natural course of acute CSC patients, even if the subretinal fluid was absorbed, OCT still showed that there was persistent PED at the leakage point, thicker Haller layer and thinner inner choroid layer. These findings provided more clues to the pathogenesis of CSC.
论著

结合多尺度特征融合的扩张残差U-Net分割网络在视网膜自动分层中的应用

Multie-scale hierarchical feature extraction combined with dilated-residual U-Net for retina automatic segmentation

:5-10
 
目的:对视网膜光学相干断层扫描图像中不同层和积液区域的分割。方法:提出一种基于深度学习的轻量级的神经网络,参考DRUNet体系、膨胀卷积和残差网络的架构,通过连接不同深度网络处得到的上采样输出,进行多尺度特征融合,使网络能够更好地识别出图像中的边界信息。结果:改进型DRUNet显著提升了视网膜分层的效果,准确率较U-Net提高了1.25%,同时能提前1~2次迭代达到传统U-Net的准确度。结论:本文采用的网络结构提高了对视网膜光学相干断层扫描图像的分割性能,同时降低了网络参数,具有强大的应用潜力。
Objective: To achieve the segmentation of different layers and fluid areas on the optical coherence tomography (OCT) image of the retina. Methods: A lightweight neural network based on deep learning was proposed. The network structure adopted in this study was designed based on the architecture of dilated-residual U-Net. By connecting the upsampling output obtained at different depth networks, multi-scale feature fusion was performed to enable the system to accurately identify the boundaries on the OCT image. Results: Compared with U-Net, this algorithm could achieve the same accuracy with 1–2 epochs less, and the accuracy was also improved by 1.25%. Conclusion: The proposed network improves the segmentation performance of retinal OCT images, and reduces the number of parameters, which demonstrates the network has great application potential.
论著

ARIA软件测量视网膜血管管径及迂曲度的重复性和再现性

Repeatability and reproducibility of retinal vessel diameter and tortuosity measured by ARIA software

:383-392
 
目的:评价不同操作者使用视网膜自动图像分析(automated retinal image analyzer,ARIA)软件测量糖尿病患者视网膜血管管径和迂曲度的重复性和再现性。方法:采用前瞻性诊断性试验研究设计,由2名熟练操作者应用ARIA软件对49名糖尿病患者以视盘为中心的45°数码眼底彩照进行视网膜血管半自动分析。通过测量距离视盘0.5~1.0个视盘直径内的视网膜动静脉血管参数,各选取6条最粗的动静脉综合计算得到中央动脉管径当量(central retinal artery equivalent,CRAE)、视网膜中央静脉管径当量(central retinal vein equivalent,CRVE)、视网膜动静脉比值(arteriole-to-venule ratio,AVR)、平均视网膜动脉迂曲度(mean retinal arteriolar tortuosity,MRAT)、平均视网膜静脉迂曲度(mean retinal venular tortuosity,MRVT)。在此基础上,评价同一操作者与不同操作者间测量结果的重复性和再现性,并绘制Bland-Altman图进行一致性分析。结果:操作者A两次测量的CRAE、CRVE、AVR差异无统计学意义(t= -0.089、-1.425、1.214,均P>0.05),重复性良好(ICC≥0.725)且高度正相关(r≥0.721)。操作者A与操作者B对同一受试者2次测量的CRAE、CRVE、AVR差异无统计学意义(t= -1.169、-0.050、0.941,均P>0.05),再现性良好(ICC≥0.750)且高度正相关( r≥0.697)。同一操作者和不同操作者间的MR AT、MRVT差异无统计学意义
(P≥0.05),Spearman非参数相关性分析重复性和再现性较好(r≥0.571,r≥0.588,均P<0.01)。结论:利用ARIA软件测量糖尿病患者视网膜血管管径及迂曲度具有良好的重复性和再现性。
Objective: To assess the repeatability and reproducibility in measurements of retinal vessel diameter and tortuosity in diabetic patients using the Automated Retinal Image Analyzer (ARIA). Methods: A prospective diagnostic trial was performed; 2 skilled researchers performed a semi-automatic analysis of retinal vessels in digital color fundus photography (45° field of view centered on the optic disc) from 49 diabetic patients using ARIA software. Six largest arterioles and venules coursing through an area of 0.5–1.0 disc diameter surrounding the optic disc were selected. Then the retinal vessel parameters were calculated into the central retinal artery equivalent (CRAE), central retinal vein equivalent (CRVE), arteriole-to-venule ratio (AVR), mean retinal arteriolar tortuosity (MRAT) and mean retinal venular tortuosity (MRVT). To evaluate the repeatability of those values in 2 measurements by the same researcher and the reproducibility between different researchers. Bland-Altman analysis was employed to assess the agreement of the outcomes. Results: There was no significant difference in the CRAE, CRVE, and AVR values in the two-time measurements of researcher A (t= -0.089, -1.425 and 1.214, all P>0.05). It also showed a good repeatability and a highly positive correlation (ICC≥0.725, r≥0.721). There was no significant difference in those measurements between researcher A and B (t= -1.169, -0.050, 0.941, all P>0.05). It also showed a good reproducibility and a highly positive correlation (ICC≥0.750, r≥0.697). Results showed that there was no significant difference in the MRAT and MRVT values both in intraobserver and interobserver measurements (P≥0.05). Besides, Spearman’s nonparametric correlation analysis showed good repeatability and reproducibility (r≥0.571, r≥0.588, all P<0.01). Conclusion: ARIA software yields good repeatability and reproducibility in the measurements of retinal vessel diameter and tortuosity in diabetic patients.
论著

EDN1和CFH的基因多态性与2型糖尿病增生性视网膜病变的相关性

Association of EDN1 and CFH gene polymorphisms with proliferative diabetic retinopathy in type 2 diabetes mellitus

:374-382
 
目的:探讨基因多态性与2型糖尿病(type 2 diabetes mellitus,T2DM)患者发生增生性糖尿病性视网膜病变(proliferative diabetic retinopathy,PDR)的相关性。方法:2019年1月至2020年9月桂林医学院附属医院收治的700名T2DM患者,分为无糖尿病性视网膜病变(non-diabetic retinopathy,NDR)组(n=386)与PDR组(n=314)。收集临床基本资料,抽取患者外周血,使用竞争性等位基因特异性聚合酶链反应(kompetitive allele specific polymerase chain reaction,KASP)基因分型检测法检测两组患者血清中的内皮素-1(endothelin 1,EDN1)基因的rs5370位点和补体因子H(complement factor H,CFH)基因的rs800292位点基因型。用logistic回归分析这2个基因位点的多态性与广西汉族T2DM患PDR的关系。结果:收缩压、使用胰岛素治疗以及肾小球滤过率(glomerular filtration rate,GFR)的分析结果显示两组间差异有统计学意义(P收缩压=0.025,P胰岛素=0.001,PGFR=0.013)。排除以上混杂因素后,EDN1基因的rs5370位点上TT基因型与PDR易感性呈正相关(P=0.03,OR=2.973;adj.P=0.011,OR=2.718);CFH基因的rs800292位点上AA基因型与PDR易感性呈正相关(P=0.037,OR=1.949;adj.P=0.044,OR=2.058)。结论:收缩压增高、GFR降低可能与T2DM患者发生PDR相关。EDN1基因的rs5370位点与CFH基因的rs800292位点的多态性与广西汉族人群的PDR易感性显著相关。
Objective: To investigate the relationship between gene-polymorphisms and proliferative diabetic retinopathy in patients who have type 2 diabetes mellitus (T2DM). Method: A total of 700 hospitalized T2DM patients were included in this study from January 2019 to September 2020. They were divided into two groups: the no-diabetic retinopathy (NDR) group (n=386) and the proliferative diabetic retinopathy (PDR) group (n=314). Basic clinical data were collected, and clinical indexes affecting diabetic retinopathy were analyzed. Two tag SNPs rs5370 in endothelin 1 (EDN1) and rs800292 in complement factor H (CFH) were examined using kompetitive allele-specific polymerase chain reaction (KASP) genotyping assays. Logistic regression was used to analyse the relationship between the polymorphisms of these two SNPs and PDR in a Guangxi Han population with T2DM. Results: Significant differences were found through the analysis of the systolic blood pressure—whether using insulin or not—and the glomerular filtration rate (GFR) between the two groups (Psystolic blood pressure=0.025, Pinsulin=0.001, PGFR=0.013) The TT genotype of rs5370 was determined to be associated with an increased risk of PDR (P=0.03, OR=2.973; adj.P=0.011, OR=2.718). The AA genotype of rs800292 was also determined to be associated with an increased risk of PDR (P=0.037, OR=1.949; adj.P=0.044, OR=2.058). Conclusion: Increased systolic blood pressure and decreased GFR may be associated with PDR in patients with T2DM. The rs5370 polymorphism of the EDN1 gene and the rs800292 polymorphism of the CFH gene are significantly associated with the risk of PDR in Guangxi’s Han population.
论著

二甲双胍治疗与2型糖尿病患者糖尿病性视网膜病变的相关性研究

Relationship between metformin treatment and diabetic retinopathy in patients with type 2 diabetes mellitus

:298-306
 
目的:探讨2型糖尿病(type 2 diabetes mellitus,T2DM)患者二甲双胍治疗与糖尿病性视网膜病变(diabetic retinopathy,DR)的相关性。方法:回顾2015年9月至2020年8月在中日友好医院眼科就诊的1 891例T2DM患者的临床资料,对病程≥10年的324例T2DM患者的一般资料、内科疾病史、糖尿病治疗史、眼科检查和实验室血生化指标进行回顾性病例研究。根据是否接受二甲双胍治疗分为二甲双胍治疗组与非二甲双胍治疗组,根据眼底检查结果同时结合DR临床诊断标准,将DR分为无明显DR、非增生性DR及增生性DR。采用logistic多因素回归分析判断年龄、性别、糖尿病发病年龄、糖尿病病程、高血压病程、高血脂病程、吸烟年数、体重指数、胰岛素治疗及空腹血糖、糖化血红蛋白、总胆固醇、三酰甘油、尿酸和血肌酐水平对结局变量的影响。结果:在DR的发病风险方面,二甲双胍治疗组与非二甲双胍治疗组的差异无统计学意义(P>0.05)。对T2DM患者DR发生及不同分期的相关变量行单因素及多因素分析,结果显示吸烟年数、空腹血糖及肌酐均与DR发病呈正相关(均P<0.05),而年龄与DR发病呈负相关(P<0.01),糖尿病发病年龄与DR发生呈显著负相关(OR=0.95,95%CI:0.92~0.98,P=0.0003)。在二甲双胍治疗的T2DM患者中,二甲双胍的疗程(OR=1.02,95%CI:0.96~1.08,P>0.05)及平均剂量(OR=1.50,95%CI:0.79~2.84,P>0.05)与DR的发生与进展均无显著相关性;女性DR发生与进展的风险较男性低(P<0.05);合并胰岛素治疗与DR发生呈明显正相关(OR=3.11,95%CI:1.59~6.07,P<0.01);吸烟年数长、糖化血红蛋白及尿酸水平高于正常范围均与DR的发生与进展呈正相关(P<0.05)。在口服二甲双胍患者中,未使用胰岛素治疗组和联合使用胰岛素组的DR发病风险有显著差异(P<0.01);而未口服二甲双胍患者中,胰岛素治疗与DR发生呈正相关(OR=12.43,95%CI:3.75~41.19,P<0.0001)。结论:病程10年以上T2DM患者中,二甲双胍治疗与DR发生与进展均无显著相关性。
Objective: To investigate the correlation between metformin therapy and diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM). Methods: The clinical data of 1 891 patients with type 2 diabetes mellitus attending the ophthalmology department of China-Japan Friendship Hospital from September 2015 to August 2020 were reviewed. A retrospective study was performed on 324 cases of these T2DM patients with disease duration ≥10 years. Medical records of all patients including general information, history of medical disease, diabetes treatment, ophthalmologic examination and blood biochemical indices were collected. According to whether metformin treatment was received or not, the patients were divided into a metformin-treated and a non-metformin-treated groups. DR is classified into non-obvious DR, non-proliferative DR and proliferative DR according to the fundus examination and the clinical diagnostic criteria of DR. Logistic multiple regression analysis was used to determine the effects of age, sex, age of DM onset, duration of DM, duration of hypertension,
duration of hyperlipidemia, years of smoking, body mass index, insulin treatment and fasting glucose, glycated hemoglobin, total cholesterol, triglycerides, uric acid and blood creatinine levels on DR. Results: There was no statistically significant difference in the risk of developing DR between the metformin-treated and non-metformin-treated groups (P>0.05). Univariate and multifactorial analyses of variables related to the occurrence and different stages of DR in patients with T2DM showed that years of smoking, fasting glucose and creatinine were positively associated with DR (P<0.05), while age was negatively associated with DR (P<0.01), and age of DM onset was significantly negatively associated with DR (OR=0.95, 95%CI: 0.92 to 0.98, P=0.0003). In T2DM patients treated with metformin, neither the duration of metformin (OR=1.02, 95%CI: 0.96 to 1.08, P>0.05) nor the mean dose(OR=1.50, 95%CI: 0.79 to 2.84, P>0.05) was significantly associated with developing DR. The risk of developing DR was lower in women than in men (P<0.05); combined insulin therapy was significantly positively correlated with the risk of DR (OR=3.11, 95%CI: 1.59 to 6.07, P<0.01); long-term smoking, glycosylated hemoglobin and uric acid levels higher than normal were positively associated with DR (P<0.05). In metformin users, there was a significant difference in the risk of developing DR between the no-insulin treatment group and the combined insulin group (P<0.01); and among patients not using metformin, insulin therapy was positively associated with the occurrence of DR (OR=12.43, 95%CI: 3.75 to 41.19, P<0.0001). Conclusion: There was no significant association between metformin treatment and DR among patients with T2DM for >10 years.
“筑梦·铸人”专题

视网膜类器官衍生物的研究应用

An overview on applications of retinal organoid derivatives

:255-270
 
视网膜退行性疾病的种类繁多、患病人口基数大,该病特征为终末期严重的视网膜细胞丢失。视网膜类器官(retinal organoid,RO)可通过3D干细胞体外分化培养技术大量获取,并拥有完整的各亚型视网膜细胞和经典的视网膜分层结构。因此,RO可作为最佳的视网膜退行性疾病建模方法之一,以便于发现潜在致病机制。目前,RO衍生物已被广泛用于视网膜细胞替代治疗的动物实验和临床研究,具体的成效参差不齐,可能的影响因素包括移植细胞数量、移植时间窗、移植工具等。随着RO相关研究的快速发展,视网膜退行性疾病在分子和个体上的诊断和治疗将进一步完善。
Retinal degenerative diseases, characterized by severe retinal cell loss at the end stage, are of various kinds and haunt vast amounts of patients. Retinal organoid (RO) with complete retinal cell subtypes and classic retinal stratification structures can be obtained in large quantities through stem cells in vitro 3D differentiation and culture method. Therefore, RO can serve as one of the best ways for retinal degenerative disease modeling to facilitate the decipherment of underlying pathogenic mechanisms. At present, RO derivatives have been widely used in animal experiments and clinical studies of retinal cell replacement therapy with varying results possibly affected by cell quantity, time window, or tools in terms of transplantation. With the booming progress of RO-related research, the diagnosis and treatment on molecular and individual level for retinal degenerative diseases will be further improved.
“筑梦·铸人”专题

视网膜神经节细胞特异的报告基因干细胞系构建及其应用

Establishment and application of a retinal ganglion cell-specific reporter stem cell line

:245-254
 
目的:建立能驱动GFP在视网膜神经节细胞(retinal ganglion cell,RGC)中特异性表达的小鼠胚胎干细胞系。方法:通过同源重组的方式建立Brn3b-GFP敲入的小鼠胚胎干细胞系(Brn3b-GFP ESC),利用3D培养将其诱导成视网膜类器官检测GFP表达的细胞特异性,再用流式细胞分选富集GFP阳性RGC,采用玻璃体腔注射的方式将GFP阳性RGC移植到健康小鼠和NMDA损伤模型小鼠眼中探索该细胞的应用价值。结果:Brn3b-GFP ESC经3D视网膜诱导培养后在RGC中特异性表达GFP,将这些GFP阳性RGC移植到两种小鼠中2周后能在所有视网膜内观察到GFP阳性细胞存活,且均能观察到有供体RGC整合到宿主视网膜RGC层。结论:本研究建立了RGC特异的报告基因干细胞系Brn3b-GFP ESC,通过将该细胞系诱导成视网膜类器官进而获得的GFP阳性RGC移植后能够整合进宿主视网膜。该细胞系的建立将为青光眼及相关疾病提供重要的研究手段和工具。
Objective: This study was designed to establish a mouse embryonic stem cell line that can drive GFP expression specifically in retinal ganglion cells (RGCs). Methods: In this study, we established a Brn3b-GFP knock-in embryonic stem cell line (Brn3b-GFP ESC) by homologous recombination. By 3D culture, we induced these cells into retinal organoids to investigate the cell-specificity of GFP expression. GFP-positive RGCs were then enriched by flow cytometry and transplanted by intravitreal injection into the eyes of healthy mice and NMDA injury model mice to explore the feasibility of a potential clinical application. Results: GFP was specifically expressed in RGCs following induction of Brn3b-GFP ESCs into 3D retinal organoids. Two weeks after these GFP-positive RGCs were transplanted into the control and injured mice, GFP-positive cells were observed in all transplanted retinas, and donor RGCs were seen to integrate into the RGC layer of the host retina. Conclusion: This study has established a retinal ganglion cell-specific reporter stem cell line Brn3b-GFP ESC. The GFP-positive RGCs obtained by inducing the cell line into retinal organoids can be integrated into the host retina after transplantation. The establishment of such a cell line will provide an important research tool for glaucoma and related diseases.
其他期刊
  • 眼科学报

    主管:中华人民共和国教育部
    主办:中山大学
    承办:中山大学中山眼科中心
    主编:林浩添
    主管:中华人民共和国教育部
    主办:中山大学
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  • Eye Science

    主管:中华人民共和国教育部
    主办:中山大学
    承办:中山大学中山眼科中心
    主编:林浩添
    主管:中华人民共和国教育部
    主办:中山大学
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