Knockdown of fibrillin-1 suppresses retina-blood barrier dysfunction by inhibiting vascular endothelial apoptosis under diabetic conditions

AIM:To investigate the effects of fibrillin-1(FBN1)deletion on the integrity of retina-blood barrier function and the apoptosis of vascular endothelial cells under diabetic conditions.METHODS:Streptozotocin(STZ)-induced diabetic mice were used to simulate the diabetic conditions of diabetic retinopathy(DR)patients,and FBN1 expression was detected in retinas from STZ-diabetic mice and controls.In the Gene Expression Omnibus(GEO)database,the GSE60436 dataset was selected to analyze FBN1 expressions in fibrovascular membranes from DR patients.Using lentivirus to knock down FBN1 levels,vascular leakage and endothelial barrier integrity were detected by Evans blue vascular permeability assay,fluorescein fundus angiography(FFA)and immunofluorescence labeled with tight junction marker in vivo.High glucose-induced monkey retinal vascular endothelial cells(RF/6A)were used to investigate effects of FBN1 on the cells in vitro.The vascular endothelial barrier integrity and apoptosis were detected by trans-endothelial electrical resistance(TEER)assay and flow cytometry,respectively.RESULTS:FBN1 mRNA expression was increased in retinas of STZ-induced diabetic mice and fibrovascular membranes of DR patients(GSE60436 datasets)using RNA-seq approach.Besides,knocking down of FBN1 by lentivirus intravitreal injection significantly inhibited the vascular leakage compared to STZ-DR group by Evans blue vascular permeability assay and FFA detection.Expressions of tight junction markers in STZ-DR mouse retinas were lower than those in the control group,and knocking down of FBN1 increased the tight junction levels.In vitro,30 mmol/L glucose could significantly inhibit viability of RF/6A cells,and FBN1 mRNA expression was increased under 30 mmol/L glucose stimulation.Down-regulation of FBN1 reduced high glucose(HG)-stimulated retinal microvascular endothelial cell permeability,increased TEER,and inhibited RF/6A cell apoptosis in vitro.CONCLUSION:The expression level of FBN1 increases in retinas and vascular endothelial cells under diabetic conditions.Down-regulation of FBN1 protects the retina of early diabetic rats from retina-blood barrier damage,reduce vascular leakage,cell apoptosis,and maintain vascular endothelial cell barrier function.

Glucagon-like peptide-1 receptor agonists rescued diabetic vascular endothelial damage through suppression of aberrant STING signaling

Glucagon-like peptide-1 receptor agonists(GLP-1 RAs)protect against diabetic cardiovascular diseases and nephropathy.However,their activity in diabetic retinopathy(DR)remains unclear.Our retrospective cohort study involving 1626 T2DM patients revealed superior efficacy of GLP-1 RAs in controlling DR compared to other glucose-lowering medications,suggesting their advantage in DR treatment.By single-cell RNA-sequencing analysis and immunostaining,we observed a high expression of GLP-1R in retinal endothelial cells,which was down-regulated under diabetic conditions.Treatment of GLP-1 RAs significantly restored the receptor expression,resulting in an improvement in retinal degeneration,vascular tortuosity,avascular vessels,and vascular integrity in diabetic mice.GO and GSEA analyses further implicated enhanced mitochondrial gene translation and mitochondrial functions by GLP-1 RAs.Additionally,the treatment attenuated STING signaling activation in retinal endothelial cells,which is typically activated by leaked mitochondrial DNA.Expression of STING mRNA was positively correlated to the levels of angiogenic and inflammatory factors in the endothelial cells of human fibrovascular membranes.Further investigation revealed that the cAMP-responsive element binding protein played a role in the GLP-1R signaling pathway on suppression of STING signaling.This study demonstrates a novel role of GLP-1 RAs in the protection of diabetic retinal vasculature by inhibiting STING-elicited inflammatory signals.

近视眼孔源性视网膜脱离患者对侧眼周边视网膜血管FFA观察

目的:观察近视眼孔源性视网膜脱离患者对侧眼周边视网膜血管FFA的图像特点及其影响因素。方法:回顾性研究。收集2018年10月至2019年8月于郑州大学第一附属医院就诊的78例(78只眼)单眼近视眼单侧孔源性视网膜脱离患者的对侧眼的临床资料。根据眼轴长度,将患者分为3组:A组,24例(24只眼)其眼轴长度为24.0~26.0 mm;B组,23例(23只眼)其眼轴长度26.1~28.0 mm;C组,31例(31只眼)其眼轴长度≥28.1 mm。所有患者均行荧光素眼底血管造影。观察周边视网膜血管情况及其可能影响因素。结果:A、B、C 3组均见周边视网膜血管荧光渗漏,3组发生率依次20.83%、26.09%、19.35%,差异无统计学意义(χ^2=0.370,P=0.831)。周边视网膜血管荧光渗漏发生率为21.79%,渗漏的发生率随着年龄增加而减少,≤45岁者发生率高于>45岁者,两组差异有统计学意义(χ^2=6.093,P=0.014)。不同性别组周边视网膜血管荧光渗漏差异无统计学意义(χ^2=0.679,P=0.410)。结论:近视者孔源性视网膜脱离的对侧眼部分出现周边视网膜血管荧光渗漏,且年龄越小出现的可能性越大,而与眼轴长度、性别无关。

渴络欣胶囊治疗中重度非增生型糖尿病视网膜病变的初步临床应用研究

目的观察中药渴络欣胶囊治疗中重度非增生型糖尿病视网膜病变(NPDR)的安全性及有效性。方法多中心、非盲、单臂、Ⅱa期临床研究。研究对象为2014年5月至2016年12月于中部战区总医院、南昌大学附属眼科医院、中国中医科学院西苑医院、中国中医科学院眼科医院检查确诊并接受渴络欣药物治疗的中重度NPDR患者,并据此分为中度NPDR组、重度NPDR组。所有患者均行最佳矫正视力(BCVA)、彩色眼底照相、光相干断层扫描、荧光素眼底血管造影以及实验室检查。在原有控制糖尿病的治疗基础上,给予患者口服渴络欣胶囊治疗,连续24周。以治疗后24周末为疗效判定时间点。对比观察两组患者基线及治疗后24周末BCVA、黄斑中心凹视网膜厚度(CMT)、6 mm直径范围内黄斑视网膜体积(TMV)、视网膜血管渗漏面积、视网膜无灌注区(RNP)面积变化。组间连续变量比较行独立样本Mann-Whitney U检验;分类资料比较行χ^(2)检验。结果共计纳入NPDR患者60例60只眼,失访9例,最终纳入51例51只眼,其中中度NPDR组、重度NPDR组分别为37、14只眼。基线时,中度NPDR组、重度NPDR组患眼BCVA分别为(80.1±6.8)、(81.4±6.3)个字母;CMT分别为(249.5±32.1)、(258.9±22.2)μm;TMV分别为(8.79±1.09)、(8.95±1.31)mm^(3);视网膜血管渗漏面积分别为(7.69±10.63)、(10.45±7.65)mm^(2);RNP面积分别为(2.48±5.74)、(10.63±20.06)mm^(2);伴糖尿病黄斑水肿(DME)分别为11(29.7%,11/37)、4(28.6%,4/14)只眼。治疗后24周,中度NPDR组、重度NPDR组患眼BCVA分别提高(1.3±5.2)、(3.2±3.0)个字母;与基线时比较,重度NPDR组差异有统计学意义(t=-3.986,P=0.033)。CMT分别为(252.1±45.6)、(269.8±57.2)μm;与基线时比较,差异均无统计学意义(t=-0.567、-0.925,P>0.05)。TMV分别为(9.96±1.16)、(10.09±1.32)mm^(3);与基线时比较,差异均无统计学意义(t=-0.996、-1.304,P>0.05)。视网膜血管渗漏面积分别减小(0.19±6.90)、(1.98±7.52)mm^(2);与基线时比较,差异均无统计学意义(t=0.168、0.983,P>0.05)。RNP面积分别为(3.01±6.47)、(10.36±19.57)mm^(2);与基线时比较,差异均无统计学意义(t=-1.267、0.553,P>0.05)。伴DME分别为8(21.6%,8/37)、3(21.4%,3/14)只眼;与基线时比较,差异有统计学意义(χ^(2)=11.919、4.571,P=0.001、0.033)。结论渴络欣胶囊可稳定或改善中重度NPDR患眼BCVA、CMT、TMV、RNP面积,降低视网膜血管渗漏面积。