Continuously increased generation of ROS in human plasma after cardiac arrest as determined by Amplex Red oxidation,Free Radical Research

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Continuously increased generation of ROS in human plasma after cardiac arrest as determined by Amplex Red oxidation
Free Radical Research ( IF 3.6 ) Pub Date : 2023-08-29 , DOI: 10.1080/10715762.2023.2250547
Muhammad Shoaib _{1,

2} , Nancy Kim _{1,

2} , Rishabh C Choudhary ₁ , Blanca Espin ₁ , Mitsuaki Nishikimi ₁ , Ann Iverson ₁ , Tsukasa Yagi ₁ , Seyedeh Shadafarin Marashi Shoshtari ₁ , Koichiro Shinozaki _{1,

2,

3} , Lance B Becker _{1,

2,

3} , Junhwan Kim _{1,

2,

3}

Affiliation

Abstract

Oxidative stress is believed to be a major cause of injury after cardiac arrest (CA). While the effects of ROS generated within tissues have been extensively investigated, the potential of plasma-generated ROS in contributing to CA pathology has not been examined. We utilized Amplex Red (AR) to measure the real time-generation of ROS in isolated plasma from human CA patients. We first used post-CA rat plasma to identify interfering factors for AR oxidation, and then applied this knowledge to analyze human plasma samples, accounting for the identified confounders. We found significantly increased AR oxidation rates lasting for 4 h in post-CA rat plasma compared to baseline. AR oxidation was unchanged with removal of horseradish peroxidase or addition of catalase. However, adding carboxylesterase inhibitors significantly decreased AR oxidation in rat plasma, which implicated increased carboxylesterase activity, not ROS leading to increased AR oxidation. AR oxidation rates were also significantly increased in human CA patient plasma compared to control and this increase persisted even with carboxylesterase inhibition, suggesting continuously increased ROS-generation within plasma post-CA in humans. The increased ROS generation may be one major source of injury post-CA that may be mitigated with antioxidative therapeutic strategies that can manage the ROS systemically generated in plasma over time.

KEY POLICY HIGHLIGHTS
We examined the potential of plasma as a source of ROS generation post-cardiac arrest
Rat cardiac arrest was used to guide the application of Amplex Red in human plasma
ROS generation in plasma is significantly increased after cardiac arrest in humans
Scavenging excessive ROS in post-resuscitation plasma may improve outcomes of patients

中文翻译：

Amplex Red 氧化测定心脏骤停后人血浆中 ROS 的产生持续增加

摘要

氧化应激被认为是心脏骤停（CA）后受伤的主要原因。虽然组织内产生的 ROS 的影响已被广泛研究，但血浆产生的 ROS 在促进 CA 病理学中的潜力尚未得到检验。我们利用 Amplex Red (AR) 测量人类 CA 患者分离血浆中 ROS 的实时生成。我们首先使用 CA 后大鼠血浆来识别 AR 氧化的干扰因素，然后应用这些知识来分析人类血浆样本，解释已识别的混杂因素。我们发现与基线相比，CA 后大鼠血浆中 AR 氧化率显着增加，持续 4 小时。去除辣根过氧化物酶或添加过氧化氢酶后，AR 氧化没有变化。然而，添加羧酸酯酶抑制剂显着降低了大鼠血浆中的 AR 氧化，这表明羧酰酯酶活性增加，而不是 ROS 导致 AR 氧化增加。与对照-E7��比，人类 CA 患者血浆中的 AR 氧化率也显着增加，并且即使在抑制羧酸酯酶的情况下，这种增加仍然持续，表明人类 CA 后血浆中 ROS 的生成持续增加。ROS 生成的增加可能是 CA 后损伤的主要来源之一，可以通过抗氧化治疗策略来减轻损伤，这些策略可以随着时间的推移控制血浆中系统性生成的 ROS。