HIF-1α mediates the protective effect of plasma extracellular particles induced by remote ischaemic preconditioning on oxidative stress injury in human umbilical vein endothelial cells.
HIF-1α mediates the protective effect of plasma extracellular particles induced by remote ischaemic preconditioning on oxidative stress injury in human umbilical vein endothelial cells.
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HIF-1α介导远程缺血预处理诱导的血浆细胞外颗粒对人脐静脉内皮细胞氧化应激损伤的保护作用。
DOI：
10.3892/etm.2021.10970
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发表时间：
2022-01
影响因子：
2.7
通讯作者：
Cheng X
中科院分区：
文献类型：
--
作者：
Wen M;Hu F;Gong Z;Huang C;Cheng X
远程缺血预处理（RIPC）被认为可以减轻心肌缺血/再灌注（I/R）损伤。本研究探讨了血浆颗粒物质，这是所谓的细胞外颗粒（EPs），并从细胞中释放的RIPC过程中，可以减少H2 O2诱导的损伤人脐静脉内皮细胞（HUVECs）。首先，EP来自进行或未进行RIPC的志愿者。为了在志愿者中诱导RIPC，血压袖带交替地充气5分钟和放气相同的持续时间，连续四个周期。HUVEC被分配到两组：i）第1组与假RIPC后来自志愿者的EP预孵育24小时，然后用H2 O2（1 mM; 6小时）处理以模拟I/R诱导的氧化应激的体内条件;和ii）第2组与RIPC后来自志愿者的EP预孵育24小时，然后用H2 O2处理。随后，EP来自接受假RIPC或RIPC和/或镉（Cd）预处理的大鼠。为了在大鼠中诱导RIPC，使用连接在大鼠腹股沟水平的血压袖带递送远程后肢预处理刺激。血压袖带交替充气5分钟和放气相同的时间段，连续四个周期。HUVEC被分为六组：i）第1组未处理; ii）第2组仅接受H2 O2处理（1 mM; 6小时）; iii）第3组与来自暴露于假RIPC的大鼠的EP预孵育24小时，然后用H2 O2处理; iv）第4组与来自接受腹膜内注射1 mg/kg Cd [缺氧的药理学抑制剂-诱导因子1-α（HIF-1α）在体内]假RIPC前180 min，然后用H2 O2处理; v）组5与暴露于RIPC的大鼠的EP预孵育24 h，然后用H2 O2处理; vi）组6与在RIPC前180 min腹腔注射1 mg/kg Cd的大鼠的EP预孵育24 h，然后用H2 O2处理。使用细胞计数试剂盒-8和乳酸脱氢酶测定法监测细胞活力和细胞毒性。通过流式细胞术和蛋白质印迹分析来评估细胞凋亡和坏死。与假RIPC志愿者相比，RIPC志愿者血浆中EP浓度显著增加。RIPC-EPs可提高H2 O2诱导的HUVECs的细胞活力，降低细胞毒性，减少细胞凋亡和坏死。此外，RIPC引起大鼠肢体肌肉组织中HIF-1α表达显著增加。RIPC-EPs的减脂作用被大鼠在RIPC前腹腔注射Cd所抵消。与未接受Cd治疗的大鼠相比，在RIPC前接受Cd治疗的大鼠血浆中沉淀的EP水平显著降低。本研究提示HIF-1α至少部分介导了血浆RIPC-EPs对HUVECs氧化应激损伤的保护作用。
Remote ischaemic preconditioning (RIPC) is considered to alleviate myocardial ischaemia/reperfusion (I/R) injury. The present study explored whether blood plasma particulate matter, which is termed extracellular particles (EPs), and is released from cells during RIPC, could reduce H2O2-induced damage in human umbilical vein endothelial cells (HUVECs). Firstly, EPs were derived from volunteers who did or did not undergo RIPC. To induce RIPC in volunteers, a blood pressure cuff was alternatively inflated for 5 min and deflated for the same duration for four successive cycles. HUVECs were assigned to two groups: i) Group 1 was preincubated for 24 h with EPs from volunteers after sham-RIPC, then treated with H2O2 (1 mM; 6 h) to mimic the in vivo conditions of I/R-induced oxidative stress; and ii) group 2 was preincubated for 24 h with EPs from volunteers after RIPC, then treated with H2O2. Subsequently, EPs were derived from rats received sham-RIPC or RIPC and/or cadmium (Cd) pre-treatment. To induce RIPC in rats, a remote hind limb preconditioning stimulus was delivered using a blood pressure cuff attached at the inguinal level of the rat. The blood pressure cuff was alternatively inflated for 5 min and deflated for the same time period for four successive cycles. HUVECs were assigned to six groups: i) Group 1 was untreated; ii) group 2 received only H2O2 treatment (1 mM; 6 h); iii) group 3 was preincubated for 24 h with EPs from rats exposed to sham-RIPC, then treated with H2O2; iv) group 4 was preincubated for 24 h with EPs from rats that received an intraperitoneal injection of 1 mg/kg Cd [a pharmacological inhibitor of hypoxia-inducible factor 1-α (HIF-1α) in vivo] 180 min before sham-RIPC, then treated with H2O2; v) group 5 was preincubated for 24 h with EPs from rats exposed to RIPC, then treated with H2O2; and vi) group 6 was preincubated for 24 h with EPs from rats that received an intraperitoneal injection of 1 mg/kg Cd 180 min before RIPC, then treated with H2O2. Cell viability and cytotoxicity were monitored using Cell Counting Kit-8 and lactate dehydrogenase assays. Cell apoptosis and necrosis were assessed via flow cytometry and western blot analysis. A notable increase in EP concentration in the plasma of volunteers after RIPC compared with that in the plasma of volunteers after sham-RIPC was observed. RIPC-associated EPs (RIPC-EPs) from volunteers could improve cell viability and reduce cytotoxicity, cell apoptosis and necrosis in HUVECs treated with H2O2 in vitro. Furthermore, RIPC caused a significant increase in HIF-1α expression in the rat limb musculature. The apoptosis-reducing effect of RIPC-EPs was demonstrated to be counteracted by an intraperitoneal injection of Cd before RIPC in rats. A significant decrease in the EP levels precipitated from the plasma of rats that received Cd treatment before RIPC was observed compared with rats that did not receive Cd treatment. The present study suggested that HIF-1α mediated at least partly the protective effect of plasma RIPC-EPs on oxidative stress injury in HUVECs.
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影响因子：
14.9
作者：
Bao L;Chen Y;Lai HT;Wu SY;Wang JE;Hatanpaa KJ;Raisanen JM;Fontenot M;Lega B;Chiang CM;Semenza GL;Wang Y;Luo W
通讯作者：
Luo W
影响因子：
13.5
作者：
Bromage, Daniel I.;Davidson, Sean M.;Yellon, Derek M.
通讯作者：
Yellon, Derek M.
影响因子：
2.1
作者：
Frey, Ulrich H.;Klaassen, Marina;Peters, Juergen
通讯作者：
Peters, Juergen
DOI：
10.1016/j.bbamcr.2010.10.019
发表时间：
2011-01-01
影响因子：
5.1
作者：
Hackenbeck, Thomas;Huber, Regina;Wiesener, Michael S.
通讯作者：
Wiesener, Michael S.
影响因子：
3.7
作者：
Hibert P;Prunier-Mirebeau D;Beseme O;Chwastyniak M;Tamareille S;Lamon D;Furber A;Pinet F;Prunier F
通讯作者：
Prunier F