[1]王艳歌,李苗,王刚,等.视网膜微血管周细胞来源的外泌体对糖尿病小鼠视网膜血管功能障碍的影响[J].眼科新进展,2023,43(8):589-593.[doi:10.13389/j.cnki.rao.2023.0119]
　WANG Yange,LI Miao,WANG Gang,et al.Effects of exosomes derived from retinal microvascular pericytes on retinal vascular dysfunction in diabetic mice[J].Recent Advances in Ophthalmology,2023,43(8):589-593.[doi:10.13389/j.cnki.rao.2023.0119]
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视网膜微血管周细胞来源的外泌体对糖尿病小鼠视网膜血管功能障碍的影响/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年8期
页码:
589-593
栏目:
实验研究
出版日期:
2023-08-05

文章信息/Info

Title:
Effects of exosomes derived from retinal microvascular pericytes on retinal vascular dysfunction in diabetic mice
作者:
王艳歌; 李苗; 王刚; 张贝贝; 史平玲; 唐贺; 魏圆梦; 杜恩明; 陶冶; 宋宗明
450003　河南省郑州市，河南省人民医院，郑州大学人民医院，河南大学人民医院，河南省立眼科医院，河南省眼科研究所
Author(s):
WANG Yange; LI Miao; WANG Gang; ZHANG Beibei; SHI Pingling; TANG He; WEI Yuanmeng; DU Enming; TAO Ye; SONG Zongming
Henan Provincial People’s Hospital, People’s Hospital of Zhengzhou University, People’s Hospital of Henan University, Henan Eye Hospital, Henan Eye Institute, Zhengzhou 450003, Henan Province, China
关键词:
糖尿病视网膜病变; 血管功能障碍; 周细胞; 内皮细胞; 外泌体
Keywords:
diabetic retinopathy;  vascular dysfunction;  pericytes;  endothelial cells;  exosomes
分类号:
R774
DOI:
10.13389/j.cnki.rao.2023.0119
文献标志码:
A
摘要:
目的　探讨视网膜微血管周细胞（RMVPCs）来源的外泌体（Exos）通过调控血管内皮细胞的生物学功能进而对糖尿病视网膜血管功能障碍的影响。
方法　通过超速离心法提取并鉴定人视网膜微血管周细胞(HRMVPCs)来源的Exos（HRMVPCs-Exos），并对Exos进行PKH67染色后，与人脐静脉血管内皮细胞(HUVECs)孵育进行内吞实验，此后，体外实验分为Control组（5.55 mmol·L-1葡萄糖）、高糖（HG）组（30.00 mmol·L-1葡萄糖）和HG+HRMVPCs-Exos组，各组HUVECs进行相应处理后，应用CCK-8、细胞迁移、流式细胞术以及成管实验等方法检测 HUVECs的增殖、迁移以及血管的生成。体内实验分为NC组、链脲佐菌素（STZ）组和STZ+HRMVPCs-Exos组，每组5只健康雄性C57BL/6J小鼠，各组小鼠进行相应处理后，进行眼底照相、OCT、荧光素眼底血管造影（FFA）、视网膜血管密度以及血管渗漏检测。
结果　HRMVPCs可分泌Exos，可以进入HUVECs中。与Control组相比，HG组和HG+HRMVPCs-Exos组HUVECs的存活率、迁移率以及血管生成数均显著减少（均为P<0.05）；与HG组相比，HG+HRMVPCs-Exos组HUVECs的存活率、迁移率以及血管生成数均显著升高（均为P<0.05）。STZ+HRMVPCs-Exos组小鼠视网膜结构破坏严重，并出现大量渗出物以及荧光素渗漏，而STZ组小鼠视网膜出现少量渗出物和微血管异常，NC组小鼠视网膜无明显病变。与NC组相比，STZ组和STZ+HRMVPCs-Exos组小鼠视网膜血管密度均显著下降（均为P<0.01），血管渗漏面积均显著增加（均为P<0.01）。与STZ组相比，STZ+HRMVPCs-Exos组小鼠视网膜血管密度显著下降（P<0.01），血管渗漏面积显著增加（P<0.01）。
结论　HRMVPCs-Exos进入HUVECs中，通过加强HG下HUVECs的生物学功能，最终加重糖尿病小鼠视网膜血管功能障碍。
Abstract:
Objective　To investigate the effects of retinal microvascular pericytes (RMVPCs)-derived exosomes (Exos) on diabetes-induced retinal vascular dysfunction by regulating the biological function of vascular endothelial cells.
Methods　Exos (HRMVPCs-Exos) derived from human retinal microvascular pericytes (HRMVPCs) were extracted by ultracentrifugation and identified. After being stained with PKH67, HRMVPCs-Exos were incubated with human umbilical vein endothelial cells (HUVECs) for the endocytosis experiment. Then the in-vitro studies were divided into the Control group (5.55 mmol·L-1 glucose), high glucose (HG) group (30.00 mmol·L-1 glucose), and HG+HRMVPCs-Exos group. The proliferation, migration and angiogenesis of HUVECs were detected by the Cell Counting Kit-8, cell migration assay, flow cytometry and tube formation assay after HRMVPCs-Exos were incubated with HUVECs for 48 h. The in-vivo studies were divided into the normal control (NC) group, streptozotocin (STZ) group and STZ+HRMVPCs-Exos group, with 5 healthy male C57BL/6J mice in each group. After the intervention, fundus photography, optical coherence tomography, fundus fluorescein angiography, and detection of retinal vascular density and vessel leakage were performed.
Results　HRMVPCs could secrete Exos, which can enter HUVECs. The survival rate, migration rate and angiogenesis of HUVECs in the HG group and HG+HRMVPCs-Exos group significantly reduced compared with those in the Control group (all P<0.05); compared with the HG group, these parameters significantly increased in the HG+HRMVPCs-Exos group (all P<0.05). The retinal structure of mice in the STZ+HRMVPCs-Exos group was seriously injured with a large amount of exudate and fluorescein leakage, the retina of mice in the STZ group showed a small amount of exudate and microvascular abnormalities, and the retina of mice in the NC group had no obvious lesions. Compared with the NC group, the retinal vascular density significantly decreased (both P<0.01) and the vascular leakage area significantly increased (both P<0.01) in the STZ group and STZ+HRMVPCs-Exos group. Compared with the STZ group, the retinal vascular density significantly decreased (P<0.01) and the vascular leakage area significantly increased (P<0.01) in the STZ+HRMVPCs-Exos group.
Conclusion　HRMVPCs-Exos can enter the HUVECs to enhance the biological function of HUVECs under HG, ultimately aggravating the retinal vascular dysfunction in diabetic mice.

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备注/Memo

备注/Memo:
国家自然科学基金青年项目（编号：82101162）；河南省重点研发与推广专项（编号：222102310061）；河南省医学科技攻关计划联合共建项目（编号：LHGJ20200062）；河南省立眼科医院基础研究专项（编号：21JCQN002，20JCZD001）
更新日期/Last Update: 2023-08-05