Department of Glaucoma and Cataract, Liuzhou Aier Eye Hospital, Affiliated Hospital of Aier Ophthalmology College of Central South University, Liuzhou, China.
Hum Exp Toxicol. 2022 Jan-Dec;41:9603271221099589. doi: 10.1177/09603271221099589.
MicroRNAs (miRNAs) have been reported to be involved in the progression of various diseases. Studying the regulatory mechanisms of miRNAs can help clinical treatment.
Diabetic retinopathy (DR) is one of the complications of diabetes. The objective of this study was to elucidate the underlying molecular mechanisms by which miR-200c-3p regulates the pyroptosis of DR cell.
Human retinal microvascular endothelial cells (HRMECs) and high glucose (HG) cultures established DR cell model in vitro. RT-qPCR is used to detect the expression level of miRNAs. CCK-8 assays and flow cytometry are used to detect apoptosis of HRMECs cell. Western blotting is used to detect cleaved caspase-3, cleaved caspase-1, and N-GSDMD proteins levels in HRMECs. The ELISA assay is used to detect the expression of IL-1β and IL-18. Predict and validate potential binding sites between miR-200c-3p and SLC30A7 by dual luciferase reporter gene analysis.
The results showed that HG caused damage to HRMECs through the pyroptosis pathway rather than the apoptosis pathway. MiR-200c-3p is highly expressed in HG induced-HRMECs, and knockdown of miR-200c-3p mitigates HG-induced HRMECs pyroptosis. MiR-200c-3p negatively targets SLC30A7 in HRMECs, and miR-200c-3p regulates pyroptosis of HG-induced HRMECs by targeting SLC30A7.
The results suggest that miR-200c-3p might be a promising interference target for DR prevention and treatment. The results of current study may provide new insights into development of therapeutic strategies for DR.
已有研究报道，微小 RNA（miRNAs）参与多种疾病的进展。研究 miRNAs 的调控机制有助于临床治疗。
糖尿病视网膜病变（DR）是糖尿病的并发症之一。本研究旨在阐明 miR-200c-3p 调控 DR 细胞焦亡的潜在分子机制。
体外建立人视网膜微血管内皮细胞（HRMECs）和高糖（HG）培养的 DR 细胞模型。采用 RT-qPCR 检测 miRNA 的表达水平。CCK-8 法和流式细胞术检测 HRMECs 细胞凋亡情况。Western blot 检测 HRMECs 中 cleaved caspase-3、cleaved caspase-1 和 N-GSDMD 蛋白水平。ELISA 法检测 IL-1β 和 IL-18 的表达。通过双荧光素酶报告基因分析预测和验证 miR-200c-3p 与 SLC30A7 之间的潜在结合位点。
结果表明，HG 通过细胞焦亡途径而非细胞凋亡途径对 HRMECs 造成损伤。HG 诱导的 HRMECs 中 miR-200c-3p 高表达，敲低 miR-200c-3p 可减轻 HG 诱导的 HRMECs 细胞焦亡。miR-200c-3p 在 HRMECs 中负向靶向 SLC30A7，miR-200c-3p 通过靶向 SLC30A7 调节 HG 诱导的 HRMECs 细胞焦亡。
研究结果提示 miR-200c-3p 可能成为 DR 预防和治疗的有前途的干预靶点。本研究结果可能为 DR 治疗策略的发展提供新的思路。
