Sobottka Ventura A C, Engelmann K, Dahinden C, Böhnke M
Augen- und Poliklinik, Inselspital, Universität Bern.
Ophthalmologe. 1998 Nov;95(11):755-9. doi: 10.1007/s003470050348.
Corneal cells are known to participate in the regulation of local inflammatory processes by secretion of cytokines. As the corneal endothelium may be exposed to endotoxin in organ culture and endotoxin is known to trigger inflammatory reactions, we investigated the effect of endotoxin from E. coli on organ cultured donor corneae with respect to autocrine and paracrine functions and the endothelial viability and density. 6 pairs of donor corneae were transferred to organ culture. Medium samples were taken prospectively from day 0 to day 20. On day 10 the medium was changed and one of each pair was incubated with 50 micrograms/ml of endotoxin while the other was immersed in standard organ culture medium. The samples were screened for IL-1, -2, -4, -5, -6, -8, -10, TNF alpha and GM-CSF by ELISA. In addition endothelial cell counts were performed at day 0, after 10 and after 20 days of organ culture, using the fixed frame technique. All endotoxin-incubated organ culture media showed significantly increased IL-6 and -8 levels compared to the fellow cornea and to pre-exposure levels (P < or = 0.004). In the endotoxin-treated corneae a significantly higher endothelial cell loss occurred (P = 0.007) and signs of degeneration were observed. None of the other cytokines (IL-1 alpha, IL-2, IL-4, IL-5, IL-10, GM-CSF and TNF alpha) were detected during either the first (1 to 10-day) or the second (11 to 20-day) phase of incubation. A correlation seems to exist between an increase in IL-6 and -8 induced by endotoxin, and endothelial cell loss in organ culture.
已知角膜细胞可通过分泌细胞因子参与局部炎症过程的调节。由于角膜内皮细胞在器官培养中可能会接触到内毒素，且已知内毒素会引发炎症反应，因此我们研究了大肠杆菌内毒素对器官培养的供体角膜自分泌和旁分泌功能、内皮细胞活力及密度的影响。将6对供体角膜转移至器官培养体系。从第0天到第20天前瞻性采集培养基样本。在第10天更换培养基，每对中的一个与50微克/毫升的内毒素一起孵育，而另一个浸泡在标准器官培养基中。通过酶联免疫吸附测定法（ELISA）检测样本中的白细胞介素-1、-2、-4、-5、-6、-8、-10、肿瘤坏死因子α和粒细胞-巨噬细胞集落刺激因子（GM-CSF）。此外，在器官培养的第0天、第10天和第20天后，采用固定框架技术进行内皮细胞计数。与对侧角膜和暴露前水平相比，所有经内毒素孵育的器官培养基中白细胞介素-6和-8水平均显著升高（P≤0.004）。在内毒素处理的角膜中，内皮细胞损失明显更高（P = 0.007），并观察到退变迹象。在孵育的第一阶段（第1至10天）或第二阶段（第11至20天）均未检测到其他细胞因子（白细胞介素-1α、白细胞介素-2、白细胞介素-4、白细胞介素-5、白细胞介素-10、GM-CSF和肿瘤坏死因子α）。内毒素诱导的白细胞介素-6和-8增加与器官培养中的内皮细胞损失之间似乎存在相关性。
