Sjolander A, Schippert A, Hammarstrom S
Department of Cell Biology, Faculty of Health Sciences University Hospital, Linkoping, Sweden.
Prostaglandins. 1993 Jan;45(1):85-96. doi: 10.1016/0090-6980(93)90092-l.
The present study was carried out to further characterize the role of non-inflammatory cells in the inflammatory process. More specifically, we have investigated whether human epithelial cells can generate inflammatory lipid mediators via activation of the 5-lipoxygenase pathway. The cells were stimulated with the calcium ionophore A23187 (5 microM) for different periods of time, after which the production of eicosanoids was determined by gradient reverse-phase high performance liquid chromatography (RP-HPLC) and rapid spectral detection, permitting continuous ultraviolet spectroscopy. In both non-prelabeled cells and cells prelabeled with [1-14C]arachidonic acid, cell stimulation for 30 min or more resulted in the production of two important 5-lipoxygenase products: 5-hydroxyeicosatetraenoic acid (5-HETE) and leukotriene B4 (LTB4). Stimulation for 15 min or less, however, led solely to the formation of 5-HETE. The identities of 5-HETE and LTB4 were confirmed by HPLC retention times and UV spectra, as well as by gas chromatography-mass spectrometry for 5-HETE and radioimmunoassay for LTB4. It can therefore be concluded that human epithelial cells in general can produce important inflammatory mediators, which suggests that epithelial cells may play a more active role in the inflammatory process than is normally assumed.
本研究旨在进一步阐明非炎症细胞在炎症过程中的作用。具体而言,我们研究了人类上皮细胞是否能通过激活5-脂氧合酶途径产生炎症性脂质介质。用钙离子载体A23187(5微摩尔)刺激细胞不同时间,之后通过梯度反相高效液相色谱(RP-HPLC)和快速光谱检测来测定类花生酸的产生,该检测可进行连续紫外光谱分析。在未预先标记的细胞和用[1-14C]花生四烯酸预先标记的细胞中,刺激细胞30分钟或更长时间均导致两种重要的5-脂氧合酶产物的产生:5-羟基二十碳四烯酸(5-HETE)和白三烯B4(LTB4)。然而,刺激15分钟或更短时间仅导致5-HETE的形成。5-HETE和LTB4的身份通过HPLC保留时间和紫外光谱以及5-HETE的气相色谱-质谱分析和LTB4的放射免疫测定得以确认。因此可以得出结论,一般来说人类上皮细胞能够产生重要的炎症介质,这表明上皮细胞在炎症过程中可能发挥比通常认为的更积极的作用。