改良皮肤窗技术用于人类急性炎症的扩展特征分析
Modified skin window technique for the extended characterisation of acute inflammation in humans.
作者信息
Marks D J B, Radulovic M, McCartney S, Bloom S, Segal A W
机构信息
Department of Medicine, University College London, London, WC1E 6JJ, UK.
出版信息
Inflamm Res. 2007 Apr;56(4):168-74. doi: 10.1007/s00011-006-6119-6.
OBJECTIVE
To modify the skin window technique for extended analysis of acute inflammatory responses in humans, and demonstrate its applicability for investigating disease.
SUBJECTS
15 healthy subjects and 5 Crohn's patients.
TREATMENT
Skin windows, created by dermal abrasion, were overlaid for various durations with filter papers saturated in saline, 100 ng/ml muramyl dipeptide (MDP) or 10 microg/ml interleukin-8 (IL-8).
METHODS
Exuded leukocytes were analyzed by microscopy, immunoblot, DNA-bound transcription factor arrays and RT-PCR. Inflammatory mediators were quantified by ELISA.
RESULTS
Infiltrating leukocytes were predominantly neutrophils. Numerous secreted mediators were detectable. MDP and IL-8 enhanced responses. Many signalling proteins were phosphorylated with differential patterns in Crohn's patients, notably PKC alpha/beta hyperphosphorylation (11.3 +/- 3.1 vs 1.2 +/- 0.9 units, P < 0.02). Activities of 44 transcription factors were detectable, and sufficient RNA isolated for expression analysis of over 400 genes.
CONCLUSIONS
The modifications enable broad characterisation of inflammatory responses and administration of exogenous immunomodulators.
目的
改进皮肤窗技术以对人类急性炎症反应进行扩展分析，并证明其在疾病研究中的适用性。
受试者
15名健康受试者和5名克罗恩病患者。
处理
通过皮肤擦伤形成皮肤窗，用饱和于生理盐水、100 ng/ml胞壁酰二肽（MDP）或10 μg/ml白细胞介素-8（IL-8）的滤纸覆盖不同时长。
方法
通过显微镜检查、免疫印迹、DNA结合转录因子阵列和逆转录聚合酶链反应（RT-PCR）分析渗出的白细胞。通过酶联免疫吸附测定（ELISA）对炎症介质进行定量。
结果
浸润的白细胞主要为中性粒细胞。可检测到多种分泌的介质。MDP和IL-8增强了反应。在克罗恩病患者中，许多信号蛋白以不同模式发生磷酸化，尤其是蛋白激酶Cα/β过度磷酸化（11.3±3.1对1.2±0.9单位，P<0.02）。可检测到44种转录因子的活性，且分离出了足够的RNA用于400多个基因的表达分析。
结论
这些改进能够广泛表征炎症反应并给予外源性免疫调节剂。
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