Szili Endre J, Hong Sung-Ha, Short Robert D
Mawson Institute, University of South Australia, Adelaide SA 5095, Australia and Wound Management Innovation Cooperative Research Centre, Australia.
Biointerphases. 2015 Jun 24;10(2):029511. doi: 10.1116/1.4918765.
The transport of plasma generated reactive oxygen species (ROS) across a simple phospholipid membrane mimic of a (real) cell was investigated. Experiments were performed in cell culture media (Dulbecco's modified Eagle's medium, DMEM), with and without 10% serum. A (broad spectrum) ROS reporter dye, 2,7-dichlorodihydrofluorescein (DCFH), was used to detect the generation of ROS by a helium (He) plasma jet in DMEM using free DCFH and with DCFH encapsulated inside phospholipid membrane vesicles dispersed in DMEM. The authors focus on the concentration and on the relative rates (arbitrary units) for oxidation of DCFH [or the appearance of the oxidized product 2,7-dichlorofluorescein (DCF)] both in solution and within vesicles. In the first 1 h following plasma exposure, the concentration of free DCF in DMEM was ~15× greater in the presence of serum (cf. to the serum-free DMEM control). The DCF in vesicles was ~2× greater in DMEM containing serum compared to the serum-free DMEM control. These data show that serum enhances plasma ROS generation in DMEM. As expected, the role of the phospholipid membrane was to reduce the rate of oxidation of the encapsulated DCFH (with and without serum). And the efficiency of ROS transport into vesicles was lower in DMEM containing serum (at 4% efficiency) when compared to serum-free DMEM (at 32% efficiency). After 1 h, the rate of DCFH oxidation was found to have significantly reduced. Based upon a synthesis of these data with results from the open literature, the authors speculate on how the components of biological fluid and cellular membranes might affect the kinetics of consumption of plasma generated ROS.
研究了血浆产生的活性氧（ROS）穿过模拟（真实）细胞的简单磷脂膜的运输情况。实验在细胞培养基（杜尔贝科改良伊格尔培养基，DMEM）中进行，添加和不添加10%血清。使用一种（广谱）ROS报告染料2,7-二氯二氢荧光素（DCFH），通过氦（He）等离子体射流在DMEM中检测ROS的产生，分别使用游离DCFH以及将DCFH封装在分散于DMEM中的磷脂膜囊泡内的情况。作者关注溶液中和囊泡内DCFH氧化的浓度和相对速率（任意单位）[或氧化产物2,7-二氯荧光素（DCF）的出现情况]。在血浆暴露后的最初1小时内，有血清存在时DMEM中游离DCF的浓度比无血清DMEM对照高约15倍。与无血清DMEM对照相比，含血清的DMEM中囊泡内的DCF高约2倍。这些数据表明血清可增强DMEM中血浆ROS的产生。正如预期的那样，磷脂膜的作用是降低封装的DCFH的氧化速率（无论有无血清）。与无血清DMEM（效率为32%）相比，含血清的DMEM中ROS转运到囊泡中的效率较低（为4%）。1小时后，发现DCFH的氧化速率显著降低。基于这些数据与公开文献结果的综合分析，作者推测生物流体和细胞膜的成分可能如何影响血浆产生的ROS的消耗动力学。
