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Abstract：
BACKGROUND: Podocyte apoptosis plays a vital role in proteinuria pathogenesis in diabetic nephropathy (DN). The regulatory relationship between long noncoding RNAs (lncRNAs) and podocyte apoptosis has recently become another research hot spot in the DN field.
OBJECTIVE: To investigate whether lncRNA protein-disulfide isomerase-associated 3 (Pdia3) could regulate podocyte apoptosis through miR-139-3p and revealed the underlying mechanism.
METHODS: Using normal glucose or high glucose (HG)-cultured podocytes, the cellular functions and exact mechanisms underlying the regulatory effects of lncRNA Pdia3 on podocyte apoptosis and endoplasmic reticulum stress (ERS) were explored. LncRNA Pdia3 and miR-139-3p expression were measured through quantitative real-time polymerase chain reaction. Relative cell viability was detected through the cell counting kit-8 colorimetric assay. The podocyte apoptosis rate in each group was measured through flow cytometry. The interaction between lncRNA Pdia3 and miR-139-3p was examined through the dual luciferase reporter assay. Finally, western blotting was performed to detect the effect of lncRNA Pdia3 on podocyte apoptosis and ERS via miR-139-3p.
RESULTS: The expression of lncRNA Pdia3 was significantly downregulated in HG-cultured podocytes. Next, lncRNA Pdia3 was involved in HG-induced podocyte apoptosis. Furthermore, the dual luciferase reporter assay confirmed the direct interaction between lncRNA Pdia3 and miR-139-3p. LncRNA Pdia3 overexpression attenuated podocyte apoptosis and ERS through miR-139-3p in HG-cultured podocytes.
CONCLUSIONS: Taken together, this study demonstrated that lncRNA Pdia3 overexpression could attenuate HG-induced podocyte apoptosis and ERS by acting as a competing endogenous RNA of miR-139-3p, which might provide a potential therapeutic target for DN.
OBJECTIVE: To investigate whether lncRNA protein-disulfide isomerase-associated 3 (Pdia3) could regulate podocyte apoptosis through miR-139-3p and revealed the underlying mechanism.
METHODS: Using normal glucose or high glucose (HG)-cultured podocytes, the cellular functions and exact mechanisms underlying the regulatory effects of lncRNA Pdia3 on podocyte apoptosis and endoplasmic reticulum stress (ERS) were explored. LncRNA Pdia3 and miR-139-3p expression were measured through quantitative real-time polymerase chain reaction. Relative cell viability was detected through the cell counting kit-8 colorimetric assay. The podocyte apoptosis rate in each group was measured through flow cytometry. The interaction between lncRNA Pdia3 and miR-139-3p was examined through the dual luciferase reporter assay. Finally, western blotting was performed to detect the effect of lncRNA Pdia3 on podocyte apoptosis and ERS via miR-139-3p.
RESULTS: The expression of lncRNA Pdia3 was significantly downregulated in HG-cultured podocytes. Next, lncRNA Pdia3 was involved in HG-induced podocyte apoptosis. Furthermore, the dual luciferase reporter assay confirmed the direct interaction between lncRNA Pdia3 and miR-139-3p. LncRNA Pdia3 overexpression attenuated podocyte apoptosis and ERS through miR-139-3p in HG-cultured podocytes.
CONCLUSIONS: Taken together, this study demonstrated that lncRNA Pdia3 overexpression could attenuate HG-induced podocyte apoptosis and ERS by acting as a competing endogenous RNA of miR-139-3p, which might provide a potential therapeutic target for DN.
摘要：
背景：足细胞凋亡在糖尿病肾病（DN）的蛋白尿发病机制中起着至关重要的作用。长链非编码RNA(lncRNAs)与足细胞凋亡的调控关系是近年来DN领域的又一研究热点。
目的：探讨lncRNA蛋白-二硫键异构酶相关3(Pdia3)是否通过miR-139-3p调控足细胞凋亡并揭示其机制。
方法：使用正常葡萄糖或高糖（HG）培养的足细胞，研究了lncRNAPdia3对足细胞凋亡和内质网应激（ERS）的调控作用的细胞功能和确切机制。通过定量实时聚合酶链反应检测LncRNAPdia3和miR-139-3p的表达。通过细胞计数试剂盒-8比色测定检测相对细胞活力。通过流式细胞术测量各组的足细胞凋亡率。lncRNAPdia3和miR-139-3p之间的相互作用通过双荧光素酶报告基因测定进行检查。最后,进行蛋白质印迹以通过miR-139-3p检测lncRNAPdia3对足细胞凋亡和ERS的影响。
结果：在HG培养的足细胞中，lncRNAPdia3的表达显著下调。接下来,lncRNAPdia3参与HG诱导的足细胞凋亡。此外，双荧光素酶报告基因测定证实了lncRNAPdia3和miR-139-3p之间的直接相互作用。LncRNAPdia3过表达通过miR-139-3p在HG培养的足细胞中减弱足细胞凋亡和ERS。
结论：综合来看，这项研究表明，lncRNAPdia3过表达可以通过充当miR-139-3p的竞争性内源性RNA来减弱HG诱导的足细胞凋亡和ERS，这可能为DN提供潜在的治疗靶点。
目的：探讨lncRNA蛋白-二硫键异构酶相关3(Pdia3)是否通过miR-139-3p调控足细胞凋亡并揭示其机制。
方法：使用正常葡萄糖或高糖（HG）培养的足细胞，研究了lncRNAPdia3对足细胞凋亡和内质网应激（ERS）的调控作用的细胞功能和确切机制。通过定量实时聚合酶链反应检测LncRNAPdia3和miR-139-3p的表达。通过细胞计数试剂盒-8比色测定检测相对细胞活力。通过流式细胞术测量各组的足细胞凋亡率。lncRNAPdia3和miR-139-3p之间的相互作用通过双荧光素酶报告基因测定进行检查。最后,进行蛋白质印迹以通过miR-139-3p检测lncRNAPdia3对足细胞凋亡和ERS的影响。
结果：在HG培养的足细胞中，lncRNAPdia3的表达显著下调。接下来,lncRNAPdia3参与HG诱导的足细胞凋亡。此外，双荧光素酶报告基因测定证实了lncRNAPdia3和miR-139-3p之间的直接相互作用。LncRNAPdia3过表达通过miR-139-3p在HG培养的足细胞中减弱足细胞凋亡和ERS。
结论：综合来看，这项研究表明，lncRNAPdia3过表达可以通过充当miR-139-3p的竞争性内源性RNA来减弱HG诱导的足细胞凋亡和ERS，这可能为DN提供潜在的治疗靶点。
