Kyle J. Riedmann, Jamie E. Meegan, Aqeela Afzal, Yatzil Cervantes-Cruz, Sarah Obeidalla, Avery M. Bogart, Lorraine B. Ware, Ciara M. Shaver, Julie A. Bastarache
{"title":"氧化无细胞血红蛋白通过激活肺微血管线粒体通透性过渡孔诱导线粒体功能障碍","authors":"Kyle J. Riedmann, Jamie E. Meegan, Aqeela Afzal, Yatzil Cervantes-Cruz, Sarah Obeidalla, Avery M. Bogart, Lorraine B. Ware, Ciara M. Shaver, Julie A. Bastarache","doi":"10.1111/micc.70012","DOIUrl":null,"url":null,"abstract":"<div>\n \n \n <section>\n \n <h3> Objective</h3>\n \n <p>Cell-free hemoglobin (CFH) is released into the circulation during sepsis where it can redox cycle from the ferrous 2+ to ferric 3+ and disrupt endothelial function, but the mechanisms of CF-mediated endothelial dysfunction are unknown. We hypothesized that oxidized CFH induces mitochondrial dysfunction via the mitochondrial permeability transition pore (mPTP) in pulmonary endothelial cells, leading to the release of mitochondrial DNA (mtDNA).</p>\n </section>\n \n <section>\n \n <h3> Methods</h3>\n \n <p>Human lung microvascular endothelial cells were treated with CFH2+/CFH3+. We measured mitochondrial mPTP activation (flow cytometry), network and mass (immunostaining), structure (electron microscopy), mtDNA release (PCR), and oxygen consumption rate (OCR; Seahorse). Plasma from critically ill patients and conditioned cell media were quantified for mtDNA and CFH.</p>\n </section>\n \n <section>\n \n <h3> Results</h3>\n \n <p>CFH3+ disrupted the mitochondrial network, activated the mPTP (1434 (874–1642) vs. 2302 (1729–2654) mean fluorescent intensity, <i>p</i> = 0.02), increased the spare respiratory capacity (30.61 (29.36–37.78) vs. 7.83 (3.715–10.63) OCR, <i>p</i> = 0.004), and caused the release of mtDNA. CFH was associated with circulating mtDNA (<i>R</i><sup>2</sup> = 0.1912, <i>p</i> = 0.0077) in plasma from critically ill patients.</p>\n </section>\n \n <section>\n \n <h3> Conclusion</h3>\n \n <p>CFH3+, not CFH2+, is the primary driver of CFH-induced lung microvascular mitochondrial dysfunction. Activation of the mPTP and the release of mtDNA are a feature of CFH3+ mediated injury.</p>\n </section>\n </div>","PeriodicalId":18459,"journal":{"name":"Microcirculation","volume":"32 4","pages":""},"PeriodicalIF":1.9000,"publicationDate":"2025-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/micc.70012","citationCount":"0","resultStr":"{\"title\":\"Oxidized Cell-Free Hemoglobin Induces Mitochondrial Dysfunction by Activation of the Mitochondrial Permeability Transition Pore in the Pulmonary Microvasculature\",\"authors\":\"Kyle J. Riedmann, Jamie E. Meegan, Aqeela Afzal, Yatzil Cervantes-Cruz, Sarah Obeidalla, Avery M. Bogart, Lorraine B. Ware, Ciara M. Shaver, Julie A. Bastarache\",\"doi\":\"10.1111/micc.70012\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div>\\n \\n \\n <section>\\n \\n <h3> Objective</h3>\\n \\n <p>Cell-free hemoglobin (CFH) is released into the circulation during sepsis where it can redox cycle from the ferrous 2+ to ferric 3+ and disrupt endothelial function, but the mechanisms of CF-mediated endothelial dysfunction are unknown. We hypothesized that oxidized CFH induces mitochondrial dysfunction via the mitochondrial permeability transition pore (mPTP) in pulmonary endothelial cells, leading to the release of mitochondrial DNA (mtDNA).</p>\\n </section>\\n \\n <section>\\n \\n <h3> Methods</h3>\\n \\n <p>Human lung microvascular endothelial cells were treated with CFH2+/CFH3+. We measured mitochondrial mPTP activation (flow cytometry), network and mass (immunostaining), structure (electron microscopy), mtDNA release (PCR), and oxygen consumption rate (OCR; Seahorse). Plasma from critically ill patients and conditioned cell media were quantified for mtDNA and CFH.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Results</h3>\\n \\n <p>CFH3+ disrupted the mitochondrial network, activated the mPTP (1434 (874–1642) vs. 2302 (1729–2654) mean fluorescent intensity, <i>p</i> = 0.02), increased the spare respiratory capacity (30.61 (29.36–37.78) vs. 7.83 (3.715–10.63) OCR, <i>p</i> = 0.004), and caused the release of mtDNA. CFH was associated with circulating mtDNA (<i>R</i><sup>2</sup> = 0.1912, <i>p</i> = 0.0077) in plasma from critically ill patients.</p>\\n </section>\\n \\n <section>\\n \\n <h3> Conclusion</h3>\\n \\n <p>CFH3+, not CFH2+, is the primary driver of CFH-induced lung microvascular mitochondrial dysfunction. 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引用次数: 0
摘要
目的:在脓毒症期间,无细胞血红蛋白(CFH)被释放到血液循环中,它可以从2+铁氧化还原循环到3+铁并破坏内皮功能,但cf介导的内皮功能障碍机制尚不清楚。我们假设氧化CFH通过肺内皮细胞的线粒体通透性过渡孔(mPTP)诱导线粒体功能障碍,导致线粒体DNA (mtDNA)的释放。方法用CFH2+/CFH3+处理人肺微血管内皮细胞。我们测量了线粒体mPTP激活(流式细胞术)、网络和质量(免疫染色)、结构(电镜)、mtDNA释放(PCR)和耗氧量(OCR);海马)。对危重患者血浆和条件细胞培养基进行mtDNA和CFH定量分析。结果CFH3+破坏线粒体网络,激活mPTP(1434(874-1642)比2302(1729-2654)平均荧光强度,p = 0.02),增加备用呼吸量(30.61(29.36-37.78)比7.83 (3.715-10.63)OCR, p = 0.004),引起mtDNA释放。CFH与危重患者血浆循环mtDNA相关(R2 = 0.1912, p = 0.0077)。结论CFH3+是cfh诱导的肺微血管线粒体功能障碍的主要驱动因子,而非CFH2+。mPTP的激活和mtDNA的释放是CFH3+介导的损伤的一个特征。
Oxidized Cell-Free Hemoglobin Induces Mitochondrial Dysfunction by Activation of the Mitochondrial Permeability Transition Pore in the Pulmonary Microvasculature
Objective
Cell-free hemoglobin (CFH) is released into the circulation during sepsis where it can redox cycle from the ferrous 2+ to ferric 3+ and disrupt endothelial function, but the mechanisms of CF-mediated endothelial dysfunction are unknown. We hypothesized that oxidized CFH induces mitochondrial dysfunction via the mitochondrial permeability transition pore (mPTP) in pulmonary endothelial cells, leading to the release of mitochondrial DNA (mtDNA).
Methods
Human lung microvascular endothelial cells were treated with CFH2+/CFH3+. We measured mitochondrial mPTP activation (flow cytometry), network and mass (immunostaining), structure (electron microscopy), mtDNA release (PCR), and oxygen consumption rate (OCR; Seahorse). Plasma from critically ill patients and conditioned cell media were quantified for mtDNA and CFH.
Results
CFH3+ disrupted the mitochondrial network, activated the mPTP (1434 (874–1642) vs. 2302 (1729–2654) mean fluorescent intensity, p = 0.02), increased the spare respiratory capacity (30.61 (29.36–37.78) vs. 7.83 (3.715–10.63) OCR, p = 0.004), and caused the release of mtDNA. CFH was associated with circulating mtDNA (R2 = 0.1912, p = 0.0077) in plasma from critically ill patients.
Conclusion
CFH3+, not CFH2+, is the primary driver of CFH-induced lung microvascular mitochondrial dysfunction. Activation of the mPTP and the release of mtDNA are a feature of CFH3+ mediated injury.
期刊介绍:
The journal features original contributions that are the result of investigations contributing significant new information relating to the vascular and lymphatic microcirculation addressed at the intact animal, organ, cellular, or molecular level. Papers describe applications of the methods of physiology, biophysics, bioengineering, genetics, cell biology, biochemistry, and molecular biology to problems in microcirculation.
Microcirculation also publishes state-of-the-art reviews that address frontier areas or new advances in technology in the fields of microcirculatory disease and function. Specific areas of interest include: Angiogenesis, growth and remodeling; Transport and exchange of gasses and solutes; Rheology and biorheology; Endothelial cell biology and metabolism; Interactions between endothelium, smooth muscle, parenchymal cells, leukocytes and platelets; Regulation of vasomotor tone; and Microvascular structures, imaging and morphometry. Papers also describe innovations in experimental techniques and instrumentation for studying all aspects of microcirculatory structure and function.