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首页> 外文期刊>Platelets >Platelets stimulate airway smooth muscle cell proliferation through mechanisms involving 5-lipoxygenase and reactive oxygen species.
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Platelets stimulate airway smooth muscle cell proliferation through mechanisms involving 5-lipoxygenase and reactive oxygen species.

机译：血小板通过涉及5-脂氧合酶和活性氧的机制刺激气道平滑肌细胞增殖。

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Continuous recruitment and inappropriate activity of platelets in the airways may contribute to airway remodeling, a characteristic feature of inflammatory airway diseases that includes increased proliferation of the smooth muscle. The aim of the present investigation was to examine the effect of platelets on proliferation of airway smooth muscle cells (ASMC) in culture and to determine the possible role of 5-lipoxygenase (5-LOX) and reactive oxygen species (ROS) in this context. ASMC obtained from guinea pigs were cultured and co-incubated with washed platelets for 24 hours. Thereafter, the proliferation was measured with the MTS-assay; the results were also verified by using thymidine incorporation, DNA measurements and manual counting. The interaction between platelets and ASMC was visualized with fluorescence microscopy. We found that platelets bind to the ASMC and the presence of platelets caused a significant dose-dependent increase in ASMC proliferation. Co-incubation of ASMC with platelets alsoincreased ROS-production, detected by the fluorescent probe DCFDA. Furthermore, the platelet-induced proliferation was reduced in the presence of the NADPH-oxidase inhibitors DPI and apocynin. A possible role of 5-LOX in platelet-induced proliferation and ROS-generation was evaluated by using the 5-LOX inhibitor AA-861 and the PLA(2)-inhibitor ATK. The results showed that inhibition of these enzymes significantly reduced the platelet-induced proliferation. Moreover, Western blot analysis revealed that the ASMC but not the platelets express 5-LOX. In addition, our experiments revealed that the presence of AA-861 and ATK significantly inhibited the ROS-production generated upon co-incubation of platelets and ASMC. In conclusion, we show that platelets have a marked capacity to induce ASMC proliferation. Furthermore, our study indicates that the interaction between platelets and ASMC leads to activation of 5-LOX in the ASMC followed by an increased ROS-production, events resulting in enhanced ASMC proliferation.The new findings are of importance in understanding possible mechanisms contributing to airway remodeling.
机译：气道中血小板的持续募集和不适当的活动可能会导致气道重塑，这是气道炎性疾病的特征，包括平滑肌的增生。本研究的目的是检查血小板对培养物中气道平滑肌细胞（ASMC）增殖的影响，并确定5-脂氧合酶（5-LOX）和活性氧（ROS）在这种情况下的可能作用。培养从豚鼠获得的ASMC，并与洗涤过的血小板共孵育24小时。之后，用MTS测定法测定增殖。通过使用胸苷掺入，DNA测量和手动计数也验证了结果。用荧光显微镜观察血小板和ASMC之间的相互作用。我们发现血小板与ASMC结合，并且血小板的存在引起ASMC增殖的明显剂量依赖性增加。通过荧光探针DCFDA检测，ASMC与血小板的共孵育还增加了ROS的产生。此外，在NADPH-氧化酶抑制剂DPI和载脂蛋白的存在下，血小板诱导的增殖减少。通过使用5-LOX抑制剂AA-861和PLA（2）抑制剂ATK评估了5-LOX在血小板诱导的增殖和ROS生成中的可能作用。结果表明，抑制这些酶显着降低了血小板诱导的增殖。而且，蛋白质印迹分析显示ASMC而不表达血小板表达5-LOX。此外，我们的实验表明，AA-861和ATK的存在显着抑制了血小板和ASMC共同孵育后产生的ROS产生。总之，我们表明血小板具有明显的诱导ASMC增殖的能力。此外，我们的研究表明血小板与ASMC之间的相互作用导致ASMC中5-LOX的活化，随后ROS产生增加，导致ASMC增殖增强的事件。新发现对理解可能导致气道的机制很重要重塑。

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