Laboratory of Respiratory Cell Biology, Dipartimento di Patologia Chirurgica, Medica, Molecolare e di Area Critica, University of Pisa and Azienda Ospedaliero-Universitaria Pisana, Ospedale di Cisanello, Via Paradisa, 2, 56124, Pisa, Italy.
Inflamm Res. 2014 Jul;63(7):539-47. doi: 10.1007/s00011-014-0723-7. Epub 2014 Mar 6.
Microparticles are membrane vesicles shed by cells upon activation and apoptosis. Agonists capable of inducing microparticle generation include cytokines, bacterial products, P-selectin, histamine. Cigarette smoke extract has also been recognized as an agonist involved in microparticle generation with an apoptosis-dependent mechanism. We investigated the possibility that cigarette smoke extract induces the rapid generation of proinflammatory microparticles by human mononuclear cells with a calcium-dependent mechanism.
Human mononuclear cells were exposed to cigarette smoke extract. [Ca(2+)]i mobilization was assessed with the fluorescent probe Fluo-4 NW. Microparticles were quantified with a prothrombinase assay and by flow cytometry. Normal human bronchial epithelial cells and A549 alveolar cells were incubated with cigarette smoke extract-induced microparticles and the generation of ICAM-1, IL-8, and MCP-1 was assessed by ELISA.
Exposure to cigarette smoke extract induced a rapid increase in [Ca(2+)]i mobilization. Microparticle generation was also increased. EGTA, verapamil and the calmodulin inhibitor, W-7, inhibited microparticle generation. Incubation of lung epithelial cells with cigarette smoke extract-induced microparticles increased the expression of proinflammatory mediators.
Exposure of mononuclear cells to cigarette smoke extract causes a rapid shedding of microparticles with a proinflammatory potential that might add to the mechanisms of disease from tobacco use.
微粒体是细胞在激活和凋亡时释放的膜泡。能够诱导微粒体生成的激动剂包括细胞因子、细菌产物、P 选择素、组氨酸。香烟烟雾提取物也被认为是一种通过凋亡依赖机制参与微粒体生成的激动剂。我们研究了香烟烟雾提取物通过钙离子依赖机制诱导人单核细胞快速产生促炎微粒体的可能性。
将人单核细胞暴露于香烟烟雾提取物中。使用荧光探针 Fluo-4 NW 评估 [Ca(2+)]i 动员。通过凝血酶原酶测定法和流式细胞术定量微粒体。用 ELISA 评估正常人类支气管上皮细胞和 A549 肺泡细胞与香烟烟雾提取物诱导的微粒体孵育后 ICAM-1、IL-8 和 MCP-1 的生成。
暴露于香烟烟雾提取物诱导 [Ca(2+)]i 动员迅速增加。微粒体生成也增加。EGTA、维拉帕米和钙调蛋白抑制剂 W-7 抑制微粒体生成。用香烟烟雾提取物诱导的微粒体孵育肺上皮细胞增加了促炎介质的表达。
单核细胞暴露于香烟烟雾提取物会导致具有促炎潜力的微粒体快速释放，这可能增加烟草使用引起疾病的机制。
