PGE2/EP2R上调内皮细胞外泌体miR-423-5p促进Müller细胞活化在糖尿病视网膜病变中的作用机制研究

Diabetic retinopathy (DR) is a common complication in patients with diabetes and has become an important cause of blindness. Latest research reports that abnormally high level of PGE2 is detected in vitreous fluid of DR patients. Our previous study found that human retinal micro-vascular endothelial cell (hRMEC) expressed 4 types of EP receptor and the level of EP2R significantly increased in high glucose-treated group. Micro-RNAs are regulatory molecules that can be packaged into exosomes and secreted from cells. A series of micro-RNAs have been related to the progress of DR, but the mechanism is unknown. Here, we show that HG-treated retinal micro-vascular endothelial cells (RMECs) secrete miR-423-5p-containing exosomes, promoting müller cells activation and VEGF secretion, which could be potently inhibited by EP2R antagonist treatment. Accordingly, we hypothesize that PGE2/EPR signaling pathway regulates the secretion of exosomes-contained miR-423-5p and participates in müller activation, retinal leakage, inflammation and neovascularization in diabetic retinopathy. To test this hypothesis, first of all, we apply rats DR model, cell experiments and clinical samples tests. Secondly, by Phoenix micron IV fundus photography system, PAS staining, Evans Blue staining, WB, immunofluorescence staining, ELISA, PCR technology, we observe the disease process, the expression or activation of key factors in the signal path. The results of the reseach will help to find some targets and theoretical basis to inhibit DR occurrence and development.

糖尿病视网膜病变（DR）是糖尿病患者常见并发症和致盲原因。DR患者玻璃体液中前列腺素E2（PGE2）水平异常升高。我们前期工作发现，PGE2受体EP2R在高糖诱导的人视网膜微血管内皮细胞（hRMECs）中变化显著。高糖处理可上调hRMECs培养上清液中外泌体包裹的miR-423-5p水平，促进Müller细胞的活化，在EP2R受体抑制剂组中逆转。据此，本课题意在研究PGE2/EP2R通路通过影响RMECs外泌体miR-423-5p，促进Müller细胞活化，参与DR视网膜血管渗漏、炎症和病理性的血管新生的机制。我们从大鼠DR模型、细胞实验和临床标本检测方面验证该假说，利用小动物活体视网膜显微成像系统、PAS染色、Evans Blue染色、WB、免疫荧光、ELISA、PCR等手段，观察疾病进程、信号通路关键因子的表达和激活情况。为抑制DR血管病变的发生与发展提供新思路和理论依据。

糖尿病视网膜病变（DR）是糖尿病患者常见并发症和致盲原因。DR患者玻璃体液中前列腺素E2（PGE2）水平异常升高。我们前期工作发现，PGE2受体EP2R在高糖诱导的人视网膜微血管内皮细胞（hRMECs）中变化显著。高糖处理可上调hRMECs培养上清液中胞外囊泡包裹的miR-423-5p水平，促进Müller细胞的活化，在EP2R受体抑制剂组中逆转。据此，本课题意在研究PGE2/EP2R通路通过影响RMECs胞外囊泡miR-423-5p，促进Müller细胞活化，参与DR视网膜血管渗漏、炎症和病理性的血管新生的机制。我们从临床标本的测序、大鼠DR模型、细胞实验和临床标本检测方面验证该假说，利用小动物活体视网膜显微成像系统、PAS染色、Evans Blue染色、WB、免疫荧光、ELISA、PCR等手段，观察疾病进程、信号通路关键因子的表达和激活情况。为抑制DR血管病变的发生与发展提供新思路和理论依据。