Introduction

Diabetic patients are often accompanied by complications of diabetic vascular disease, which could lead to heart failure or stroke. In this work, we explored the role of miR-503/Apelin-12 in diabetic angiopathy (DA) in vitro.

Methods

ELISA and qPCR were applied to assess the expression of miR-503 and Apelin-12 in high glucose (HG)-treated microvascular endothelial cells (HMEC-1). The effects of miR-503 on apoptosis, inflammation and oxidative stress were assessed by flow cytometry, western blotting, qPCR, and ELISA. The interaction between miR-503 and Apelin-12 was evaluated by dual-luciferase reporter assay, qPCR and ELISA, respectively. Western blotting was performed to examine the function of miR-503/Apelin-12 on JNK and p38MAPK activation.

Results

MiR-503 was markedly increased and Apelin-12 was decreased in HG-treated HMEC-1 cells. MiR-503 inhibitor significantly assuaged apoptosis, inflammation and oxidative stress in HMEC-1 cells. MiR-503 could specifically bind to the 3′UTR of Apelin and inversely downregulate Apelin-12 expression. Furthermore, Apelin-12 suppressed apoptosis, inflammation and oxidative stress. Inhibition of Apelin-12 could partially reverse the decrease of p-JNK and p-p38 expression levels induced by miR-503 suppression.

Conclusion

In HG-induced microvascular cells injury, miR-503/Apelin-12 enhances inflammation and oxidative stress by regulating JNK and p38MAPK pathway, suggesting a potential therapeutic target for DA.

中文翻译：

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miR-503/Apelin-12 通过 JNK 和 p38MAPK 信号通路介导高糖诱导的微血管内皮细胞损伤。

介绍

糖尿病患者通常伴有糖尿病血管疾病的并发症，这可能导致心力衰竭或中风。在这项工作中，我们在体外探索了 miR-503/Apelin-12 在糖尿病血管病 (DA)中的作用。

方法

应用 ELISA 和 qPCR 评估高糖 (HG) 处理的微血管内皮细胞 (HMEC-1) 中 miR-503 和 Apelin-12 的表达。通过流式细胞术、蛋白质印迹、qPCR 和 ELISA 评估 miR-503 对细胞凋亡、炎症和氧化应激的影响。分别通过双荧光素酶报告基因测定、qPCR 和 ELISA 评估 miR-503 和 Apelin-12 之间的相互作用。进行蛋白质印迹以检查 miR-503/Apelin-12 对 JNK 和 p38MAPK 激活的功能。

结果

在 HG 处理的 HMEC-1 细胞中，MiR-503 显着增加，Apelin-12 减少。MiR-503 抑制剂显着减轻 HMEC-1 细胞的凋亡、炎症和氧化应激。MiR-503 可以特异性结合 Apelin 的 3'UTR 并反向下调 Apelin-12 的表达。此外，Apelin-12 抑制细胞凋亡、炎症和氧化应激。抑制 Apelin-12 可以部分逆转由 miR-503 抑制诱导的 p-JNK 和 p-p38 表达水平的降低。

结论

在 HG 诱导的微血管细胞损伤中，miR-503/Apelin-12 通过调节 JNK 和 p38MAPK 通路增强炎症和氧化应激，提示 DA 的潜在治疗靶点。