Cytotoxicity of N-dodecanoyl-L-homoserine lactone and 5-N-dodecyl resorcinol to human granulocytes and monocytes: a comparative study
Cytotoxicity of N-dodecanoyl-L-homoserine lactone and 5-N-dodecyl resorcinol to human granulocytes and monocytes: a comparative study
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N-十二酰-L-高丝氨酸内酯和5-N-十二烷基间苯二酚对人粒细胞和单核细胞的细胞毒性：比较研究
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
V. Chereshnev
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作者：
T. Sviridova;D. Deryabin;Olga Cyganok;V. Chereshnev
源自微生物的小分子 (SMOM) 在细菌与细菌之间的通讯以及细菌与其宿主之间的相互作用中发挥着重要作用。本研究的目的是比较分析两种结构接近的 SMOM：N-十二酰基-L-高丝氨酸内酯 (C12-HSL) 和 5-N-十二烷基间苯二酚 (C12-AR) 对人类粒细胞和单核细胞的细胞毒性活性。通过台盼蓝染料排除法测定细胞活力。为了区分细胞死亡机制，进行了坏死和凋亡测试。通过 caspase-3 活性和组蛋白相关 DNA 片段的水平来鉴定经历凋亡的细胞。为了评估细胞裂解，使用乳酸脱氢酶释放测试。此外，还研究了 SMOM 对红细胞膜稳定性的作用。与单核细胞相比，所研究的微摩尔浓度的 SMOM 对粒细胞具有更大的剂量依赖性细胞毒性，但它们使用不同的机制来影响细胞死亡途径。 C12-HSL 特异性诱导细胞凋亡，类似于先前报道的源自铜绿假单胞菌的 3-oxoN-十二酰基-L-高丝氨酸内酯的活性。相比之下，C12AR 诱导快速细胞溶解作用（坏死），如细胞质乳酸脱氢酶的释放所示，并且可能是由细胞膜不稳定所定义的。我们的数据表明，C12-HSL 和 C12-AR 都可以消除关键的防御细胞，否则这些细胞将参与破坏病原菌。这些结果强化了 SMOM 双功能概念，即此类细菌分子不仅调节细菌与细菌之间的相互作用，而且还打破免疫防御，成为宿主逃避的一种新的重要机制。
Small molecules originating from microbes (SMOMs) play a significant role in bacterial-bacterial communication and in interactions between bacteria and their hosts. The aim of this study is a comparative analysis of the cytotoxic activity to human granulocytes and monocytes of two structurally close SMOMs: N-dodecanoyl-L-homoserine lactone (C12-HSL) and 5-N-dodecyl resorcinol (C12-AR). Cell viability was determined by Trypan blue dye exclusion. To distinguish cell death mechanisms, both necrosis and apoptosis tests were carried out. Cells undergoing apoptosis were identified by caspase-3 activity and the level of histone-associated DNA fragments. To evaluate cell lysis, the lactate dehydrogenase release test was used. In addition, the SMOM’s action on erythrocyte membrane stability was investigated. The investigated SMOMs in micromolar concentrations showed more dose-dependent cytotoxicity against granulocytes than monocytes, but they used different mechanisms to impinge on the cell death pathway. C12-HSL specifically induced apoptosis similar to the activity previously reported for 3-oxoN-dodecanoyl-L-homoserine lactone that originated from Pseudomonas aeruginosa. In contrast, C12AR induced fast cytolytic effects (necrosis) as shown by the release of cytoplasmic lactate dehydrogenase, and presumably were defined by cellular membrane destabilisation. Our data demonstrate that both C12-HSL and C12-AR can eliminate key defence cells, which would otherwise participate in the destruction of pathogenic bacteria. These results reinforce the SMOMs bifunctionality concept that such bacterial molecules not only regulate bacterial-bacterial interactions but also break immune defences as a new and important mechanism of host evasion.