HIF-1α/JMJD1A signaling regulates inflammation and oxidative stress following hyperglycemia and hypoxia-induced vascular cell injury.
HIF-1α/JMJD1A signaling regulates inflammation and oxidative stress following hyperglycemia and hypoxia-induced vascular cell injury.
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HIF-1α/JMJD1A 信号调节高血糖和缺氧诱导的血管细胞损伤后的炎症和氧化应激
DOI：
10.1186/s11658-021-00283-8
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发表时间：
2021-09-03
影响因子：
8.3
通讯作者：
Wang M
中科院分区：
文献类型：
--
作者：
Zhao M;Wang S;Zuo A;Zhang J;Wen W;Jiang W;Chen H;Liang D;Sun J;Wang M
内皮细胞（EC）损伤加速糖尿病大血管并发症的进展。缺氧是EC损伤的重要原因。缺氧诱导因子-1 α（Hypoxia-inducible factor-1 alpha，HIF-1α）是一种重要的缺氧调节蛋白。我们前期的研究表明，高糖低氧条件下可上调HIF-1α的表达，增强EC的炎症损伤，且不依赖于核因子-κ B（NF-κB）通路。然而，目前尚不清楚HIF-1α是否通过表观遗传相关机制在血管疾病中发挥作用。我们利用RNA测序（RNA-seq）和小干扰HIF-1α（si-HIF-1α）技术对高糖和低氧诱导的人脐静脉内皮细胞（HUVECs）进行基因表达分析和分子机制研究。采用定量逆转录-聚合酶链反应（qRT-PCR）和Western blot检测HIF-1α和Jumonji结构域蛋白1A（JMJD 1A）的表达，酶联免疫吸附试验（ELISA）检测细胞上清液中炎性蛋白的分泌，染色质免疫沉淀（Ch-IP）检测HIF-1α和JMJD 1A之间的蛋白相互作用。我们使用细胞计数试剂盒8（CCK-8）分析细胞活力，并通过使用检测试剂盒和流式细胞术评估氧化应激指标。高糖低氧可上调HIF-1α表达，下调HIF-1α表达可降低HUVECs炎症反应和氧化应激水平。为了确定下游通路，我们通过RNA-sEq观察了组蛋白去甲基化酶基因和相关通路，其中JMJD 1A是组蛋白去甲基化酶中上调最多的基因。此外，我们还观察到HIF-1α与JMJD 1A启动子结合，并且si-HIF-1α对高糖和缺氧诱导的HUVECs氧化应激和炎性细胞因子的改善作用可被JMJD 1A过表达逆转。此外，JMJD 1A的敲低降低了炎症和氧化应激损伤。为了确定JMJD 1A相关因素，我们对JMJD 1A敲低的HUVECs进行基因表达分析。我们观察到，炎症下调和氧化应激途径丰富，FOS和FOSB可能是重要的保护性转录因子。这些发现为HIF-1α/JMJD 1A信号通路参与高糖缺氧诱导的HUVECs炎症和氧化应激提供了新的证据。此外，这一途径可能作为一种新的调节剂的氧化应激和炎症相关的事件，在糖尿病血管损伤，从而有助于糖尿病和血管疾病的病理进展。在线版本包含补充材料，可通过10.1186/s11658-021-00283-8获得。
Endothelial cell (EC) injury accelerates the progression of diabetic macrovascular complications. Hypoxia is an important cause of EC injury. Hypoxia-inducible factor-1 alpha (HIF-1α) is an important hypoxia regulatory protein. Our previous studies showed that high-glucose and hypoxic conditions could upregulate HIF-1α expression and enhance EC inflammatory injury, independently of the nuclear factor kappa-B (NF-κB) pathway. However, it is not clear whether HIF-1α plays a role in vascular disease through epigenetic-related mechanisms. We conducted gene expression analysis and molecular mechanistic studies in human umbilical vein endothelial cells (HUVECs) induced by hyperglycemia and hypoxia using RNA sequencing (RNA-seq) and small interfering HIF-1α (si-HIF-1α). We determined HIF-1α and Jumonji domain-containing protein 1 A (JMJD1A) expression by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot, analyzed inflammatory protein secretion in the cell supernatant by enzymelinked immunosorbent assay (ELISA), and assessed protein interaction between HIF-1α and JMJD1A by chromatin immunoprecipitation (Ch-IP). We used the Cell Counting Kit8 (CCK-8) assay to analyze cell viability, and assessed oxidative stress indicators by using a detection kit and flow cytometry. High glucose and hypoxia up-regulated HIF-1α expression, and down-regulated HIF-1α decreased the level of inflammation and oxidative stress in HUVECs. To determine the downstream pathways, we observed histone demethylases genes and related pathway by RNA-sEq. Among these, JMJD1A was the most upregulated gene in histone demethylases. Moreover, we observed that HIF-1α bound to the promoter of JMJD1A, and the ameliorative effects of si-HIF-1α on oxidative stress and inflammatory cytokines in high-glucose and hypoxia-induced HUVECs were reversed by JMJD1A overexpression. Furthermore, knockdown of JMJD1A decreased inflammatory and oxidative stress injury. To determine the JMJD1A-related factors, we conducted gene expression analysis on JMJD1A-knockdown HUVECs. We observed that downregulation of inflammation and the oxidative stress pathway were enriched and FOS and FOSB might be important protective transcription factors. These findings provide novel evidence that the HIF-1α/JMJD1A signaling pathway is involved in inflammation and oxidative stress in HUVECs induced by high glucose and hypoxia. Also, this pathway might act as a novel regulator of oxidative stress and inflammatory-related events in response to diabetic vascular injury and thus contribute to the pathological progression of diabetes and vascular disease. The online version contains supplementary material available at 10.1186/s11658-021-00283-8.
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