circ-PRKCB acts as a ceRNA to regulate p66Shc-mediated oxidative stress in intestinal ischemia/reperfusion
circ-PRKCB acts as a ceRNA to regulate p66Shc-mediated oxidative stress in intestinal ischemia/reperfusion
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circ-PRKCB 作为 ceRNA 调节肠道缺血/再灌注中 p66Shc 介导的氧化应激
DOI：
10.7150/thno.44250
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Tian, Xiaofeng
中科院分区：
文献类型：
--
作者：
Feng, Dongcheng;Wang, Zhecheng;Tian, Xiaofeng
背景：氧化应激在肠缺血/再灌注（I/R）损伤的发病机制中起重要作用。衔接蛋白p66Shc是肠道中活性氧（ROS）生成的关键调节因子和I/R损伤的介质，但在肠道I/R过程中直接调节p66Shc表达的上游机制仍不清楚。最近的研究表明，非编码rna，如环状rna (circRNAs)，在生理和病理过程中是重要的参与者，基于它们在基因表达中的多种调节作用。本研究旨在阐明p66Shc在肠道I/R氧化损伤中的作用，并探讨环状rna海绵对p66Shc的调节作用。方法：采用肠系膜上动脉（SMA）闭塞法诱导小鼠肠I/R。I/R激发前静脉注射miR-339-5p agomir或circ-protein kinase C β (PRKCB) siRNA。此外，在体外对Caco-2细胞进行缺氧/再氧化（H/R）模拟体内I/R模型。结果：在体外，p66Shc缺乏通过降低线粒体超氧阴离子（O-2(-)）水平、抑制NADPH氧化酶活性和增强抗氧化酶表达，显著降低H/ r诱导的ROS过量产生。此外，miR-339-5p被鉴定为直接调节p66Shc在肠道中的表达。此外，我们发现从PRKCB基因转录的circRNA，称为circ-PRKCB，作为内源性miR-339-5p海绵调节p66Shc的表达。circ-PRKCB沉默或miR-339-5p过表达可显著下调p66Shc表达，减轻体内和体外氧化应激水平和I/R损伤。值得注意的是，与小鼠肠道I/R相关的circ-PRKCB水平升高和miR-339-5p水平降低与肠梗死患者一致。结论：我们的研究结果揭示了circ-PRKCB/miR-339-5p/p66Shc信号通路在调节I/R肠氧化应激中的关键作用。该途径可能是肠I/R损伤的潜在治疗靶点。
Background: Oxidative stress has emerged as an essential factor in the pathogenesis of intestinal ischemia/reperfusion (I/R) injury. The adaptor protein p66Shc is a key regulator of reactive oxygen species (ROS) generation and a mediator of I/R damage in the intestine, but the upstream mechanisms that directly regulate p66Shc expression during intestinal I/R remain largely unknown. Recent studies have suggested that noncoding RNAs, such as circular RNAs (circRNAs), are important players in physiological and pathological processes based on their versatile regulatory roles in gene expression. The aim of this study was to elucidate the contribution of p66Shc to oxidative damage in intestinal I/R and to investigate the regulation of p66Shc by circRNA sponges.Methods: Intestinal I/R was induced in mice via superior mesenteric artery (SMA) occlusion. A miR-339-5p agomir or circ-protein kinase C beta (PRKCB) siRNA was injected intravenously before I/R challenge. In addition, Caco-2 cells were subjected to hypoxia/reoxygenation (H/R) in vitro to simulate an in vivo I/R model.Results: In vitro, p66Shc deficiency significantly reduced H/R-induced ROS overproduction by attenuating mitochondrial superoxide anion (O-2(-)) levels, suppressing NADPH oxidase activity and enhancing antioxidant enzyme expression. Moreover, miR-339-5p was identified to directly regulate p66Shc expression in the intestine. Furthermore, we found that a circRNA transcribed from the PRKCB gene, named circ-PRKCB, acted as an endogenous miR-339-5p sponge to regulate p66Shc expression. circ-PRKCB silencing or miR-339-5p overexpression significantly downregulated p66Shc expression and attenuated oxidative stress levels and I/R injury in vivo and in vitro. Notably, the increased circ-PRKCB levels and decreased miR-339-5p levels associated with murine intestinal I/R were consistent with those in patients with intestinal infarction.Conclusions: Our findings reveal a crucial role for the circ-PRKCB/miR-339-5p/p66Shc signaling pathway in regulating oxidative stress in the I/R intestine. This pathway may be a potential therapeutic target for intestinal I/R injury.