Flow-cytometric Analysis of Reactive Oxygen Species in Blood Cells: A Potential Tool for Predicting Restenosis - Insights from a Cohort Study.

Rakesh Raman Patyar, Sazal Patyar, Yash Paul Sharma, Krishan Lal Khanduja
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Abstract

Introduction: In-stent restenosis (ISR) is a recurrence of a blockage in a section of the coronary artery that has previously been treated with a stent. Molecular/biochemical pathways underlying ISR are not fully understood, but inflammation and reactive oxygen species (ROS) induced oxidative stress play a significant role in the pathogenesis of restenosis. As blood cells are highly sensitive to oxidative stress and blood is readily accessible compared to other tissues, the current study flow cytometrically investigated intracellular ROS and cytokine profile of blood cells as possible markers of restenosis. Flow cytometry is commonly used for detecting ROS and analyzing oxidative stress but so far, it has not been utilized for prediction of ISR. So, the aim of the study was to explore the potential of flow cytometric assessment of ROS levels in the blood cells as predictor of ISR.

Methods: The study was carried out in a total of 60 patients who had previously undergone coronary artery stent implantation. They were categorized as Group I - Coronary stent implanted patients without restenosis (n=30) and Group II - Coronary stent implanted patients with restenosis (n=30). Sociodemographics, biochemical and angiographic characteristics were assessed. Intracellular ROS and cytokine estimation in blood cells was done by using flow cytometric analysis.

Results: Flow cytometric measurements demonstrated a 1.3-fold increase in ROS levels in red blood cells (RBCs) and 2-fold increase in ROS levels in leucocytes in group II as compared to group I. Mean serum concentrations of pro-inflammatory cytokines: tumor necrosis factor-α (33.54 ± 6.48 vs. 20.10 ± 5.61, p <0.001***), interferon-gamma (21.76 ± 4.46 vs. 20.10 ± 5.61, p <0.001***), interleukin 6 (152.56 ± 30.67 vs. 113.95 ± 23.38, p <0.001***) were found to be higher in restenotic patients as compared to the non-restenotic patients. Correlation analysis showed that intracellular ROS levels of RBCs exhibited a significant positive correlation with late lumen loss in restenotic (r=0.71, p <0.01) as well as non-restenotic patients (r=0.59, p <0.01). Similarly, intracellular ROS levels of WBCs exhibited a significant positive correlation with late lumen loss in restenotic (r=0.72, p <0.01) as well as non-restenotic patients (r=0.61, p <0.01).

Conclusion: This study highlights the role of increased levels of intracellular ROS in blood cells in the subsequent development of ISR, which can be detected flow cytometrically. The study suggests that intracellular ROS estimation in blood cells may serve as a potential marker for restenosis and their flow cytometric analysis may facilitate the prediction of ISR.

血细胞中活性氧的流式计量分析:预测再狭窄的潜在工具--一项队列研究的启示。
导言:支架内再狭窄(ISR)是指先前用支架治疗过的冠状动脉阻塞部位再次发生堵塞。ISR的分子/生化途径尚不完全清楚,但炎症和活性氧(ROS)诱导的氧化应激在再狭窄的发病机制中起着重要作用。由于血细胞对氧化应激高度敏感,而且与其他组织相比,血液易于获取,因此本研究用流式细胞术研究了血细胞的细胞内 ROS 和细胞因子谱,以此作为再狭窄的可能标志物。流式细胞术常用于检测 ROS 和分析氧化应激,但迄今为止还未被用于预测 ISR。因此,本研究旨在探索流式细胞术评估血细胞中 ROS 水平作为 ISR 预测指标的潜力:研究对象为 60 名曾接受过冠状动脉支架植入术的患者。他们被分为 I 组--未发生再狭窄的冠状动脉支架植入患者(30 人)和 II 组--发生再狭窄的冠状动脉支架植入患者(30 人)。对社会人口学、生化和血管造影特征进行了评估。通过流式细胞分析法对血细胞内的 ROS 和细胞因子进行了评估:流式细胞仪测量显示,与第一组相比,第二组红细胞(RBC)中的 ROS 水平增加了 1.3 倍,白细胞中的 ROS 水平增加了 2 倍:本研究强调了血细胞中细胞内 ROS 水平的增加在随后的 ISR 发展中的作用,这可以通过流式细胞仪检测到。研究表明,血细胞中细胞内 ROS 的估计值可作为再狭窄的潜在标志物,对其进行流式细胞术分析有助于预测 ISR。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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