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Comparative analysis of co-culture and monoculture models in simulating diabetic neurovascular dysfunction: insights into diabetic retinopathy

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收录情况： ◇ SCIE

作者：

机构： [1]Beijing Tongren Eye Center, Tongren Hospital, Capital Medical University, Beijing, China. [2]Beijing Retinal and Choroidal Vascular Disorders Study Group, Beijing, China. [3]Clinical Research Center, Tongren Hospital, Capital Medical University, Beijing, China. [4]Save Sight Institute, Department of Ophthalmology, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia.

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关键词： diabetic neurovascular dysfunction retinal vascular endothelium cells ganglion cells co-culture monoculture

摘要：

Interaction between retinal vascular endothelial cells and neurons plays a critical role in the pathogenesis of diabetic retinopathy (DR). This study aims to compare an in vitro model over a monoculture model to simulate the neurovascular coupling under the hyperglycemic microenvironment of diabetes.Rat retinal vascular endothelial cells (RRMECs) and ganglion cells (RGCs) were seeded mono- or co-cultured in a normal (NG, 5.5 mM) and high (HG, 75 mM) glucose concentrations culture medium. Cell viability was detected by the cell counting kit-8 (CCK-8) assay. The ability of migration and lumen formation of RRMECs were determined by scratch wound, transwell migration, and lumen formation assays. The apoptosis index of cells was calculated and detected by propidium iodide (PI)/Hoechst staining. Quantitative and morphological analysis of RGCs was performed through the labeling of RGCs by brain-specific homeobox/POU domain protein 3A (BRN3A) and anti-beta-III tubulin (TUJ1). The gene and protein expression levels of occludin (OCLN) and zonula occludens-1 (ZO-1) were evaluated by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assay.The viability, migration, and lumen formation abilities of RRMECs in the HG group significantly increased (P<0.05) in both mono- and co-culture models. Migration and lumen formation abilities of RRMECs in the co-culture with HG were lower than that in the monoculture group (P<0.05). The viability of RGCs cells with HG significantly decreased in both mono- and co-culture models (Pmono<0.001, Pco<0.001), the apoptosis index of RGCs in the co-culture with HG was higher than that in the monoculture (P=0.010). The protein and gene expression of OCLN, and ZO-1 in RRMECs significantly decreased with HG culture medium in both culture models (P<0.05). In the HG group, the protein and gene expression level of the ZO-1 and OCLN of RRMECs significantly decreased in the co-culture model than that in the monoculture model (P<0.05).Compared with mono cell culture, the established co-culture in vitro system for diabetic neurovascular dysfunction can better stimulate the micro-environment of the retinal neurovascular unit.Copyright © 2023 Wang, Qiao, Kang, Zhu and Zhang.

基金：

基金编号： Grant 2016YFC1305604

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出版当年[2022]版：

大类 | 2 区医学

小类 | 2 区内分泌学与代谢

最新[2023]版：

大类 | 2 区医学

小类 | 2 区内分泌学与代谢

JCR分区：

出版当年[2021]版：

Q1 ENDOCRINOLOGY & METABOLISM

最新[2023]版：

Q2 ENDOCRINOLOGY & METABOLISM

影响因子： 3.9 最新[2023版] 4.9 最新五年平均 6.055 出版当年[2021版] 6.335 出版当年五年平均 5.555 出版前一年[2020版] 5.2 出版后一年[2022版]

第一作者：

第一作者机构： [1]Beijing Tongren Eye Center, Tongren Hospital, Capital Medical University, Beijing, China. [2]Beijing Retinal and Choroidal Vascular Disorders Study Group, Beijing, China.

通讯作者：

通讯机构： [1]Beijing Tongren Eye Center, Tongren Hospital, Capital Medical University, Beijing, China. [2]Beijing Retinal and Choroidal Vascular Disorders Study Group, Beijing, China.

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Comparative analysis of co-culture and monoculture models in simulating diabetic neurovascular dysfunction: insights into diabetic retinopathy

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