Schäper Christoph, Gläser Sven, Groneberg David A, Kunkel Gert, Ewert Ralf, Noga Oliver
Division of Cardiology and Pneumology, University of Greifswald, Friedrich-Loeffler-Strasse 23 a, 17475 Greifswald, Germany.
Regul Pept. 2009 Oct 9;157(1-3):3-7. doi: 10.1016/j.regpep.2009.07.004. Epub 2009 Jul 19.
Neurotrophins are involved in inflammatory pathways influencing several cells in healthy states and in diseases such as bronchial asthma. Recent studies have shown that nerve growth factor (NGF) is expressed in various non-neuronal cells. Furthermore, little is known about the different origins and regulation of NGF. In the present study, the expression of NGF and its regulation by dexamethasone was investigated in cultured human smooth muscle cells derived from umbilical veins (HSMC) and human iliacal arteries (HISMC).
Vascular smooth muscle cells were prepared. The presence of NGF was demonstrated by APAAP staining, western blotting, ELISA, and reverse transcription polymerase chain reaction. Vascular smooth muscle cells were incubated with dexamethasone, and cells and supernatants were collected for the measurement of NGF.
Vascular smooth muscle cells demonstrate mRNA for NGF. Proteins were detectable by western blot, ELISA, and APAAP staining. NGF Protein and mRNA were suppressed after incubation with dexamethasone (0.1 microM) for 48 h in the vascular smooth muscle cells. NGF protein was also detected in cell supernatant and was suppressed by dexamethasone as well.
These data indicate that vascular smooth muscle cells are a source of circulating NGF and thus may be involved in inflammatory responses mediated by neurotrophins. The suppression of NGF synthesis by dexamethasone might be a hint of further anti-inflammatory mechanisms of glucocorticoids.
神经营养因子参与炎症通路,在健康状态以及诸如支气管哮喘等疾病中影响多种细胞。最近的研究表明,神经生长因子(NGF)在各种非神经元细胞中表达。此外,关于NGF的不同来源和调控知之甚少。在本研究中,我们调查了从人脐静脉(HSMC)和人髂动脉(HISMC)分离培养的人平滑肌细胞中NGF的表达及其受地塞米松的调控情况。
制备血管平滑肌细胞。通过碱性磷酸酶抗碱性磷酸酶(APAAP)染色、蛋白质印迹法、酶联免疫吸附测定(ELISA)以及逆转录聚合酶链反应来证实NGF的存在。将血管平滑肌细胞与地塞米松一起孵育,收集细胞和上清液以测定NGF。
血管平滑肌细胞显示有NGF的信使核糖核酸(mRNA)。通过蛋白质印迹法、ELISA和APAAP染色可检测到蛋白质。在血管平滑肌细胞中,与地塞米松(0.1微摩尔)一起孵育48小时后,NGF蛋白质和mRNA受到抑制。在细胞上清液中也检测到了NGF蛋白质,并且它也受到地塞米松的抑制。
这些数据表明血管平滑肌细胞是循环中NGF的一个来源,因此可能参与神经营养因子介导的炎症反应。地塞米松对NGF合成的抑制可能暗示了糖皮质激素进一步的抗炎机制。