Sabourin C L, Petrali J P, Casillas R P
Division of Environmental Health Sciences, School of Public Health, The Ohio State University, Columbus 43210, USA.
J Biochem Mol Toxicol. 2000;14(6):291-302. doi: 10.1002/1099-0461(2000)14:6<291::AID-JBT1>3.0.CO;2-B.
Cutaneous exposure to sulfur mustard (bis(2-chloroethyl) sulfide, HD), a chemical warfare agent, produces a delayed inflammatory skin response and severe tissue injury. Despite defined roles of inflammatory cytokines produced or released in response to skin-damaging chemicals, in vivo cytokine responses associated with HD-induced skin pathogenesis are not well understood. Additionally, there is little information on the in vivo temporal sequence of gene expression of cytokines postexposure to HD. The goal of these studies was to identify in vivo molecular biomarkers of HD skin injury within 24 hours after HD challenge. Gene expression of interleukin 1beta (IL-1beta), granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin 6 (IL-6), and interleukin 1alpha (IL-1alpha) in the mouse ear vesicant model was examined by quantitative reverse transcription-polymerase chain reaction (RT-PCR). An increase in IL-1beta mRNA levels was first observed at 3 hours. IL-1beta, GM-CSF, and IL-6 mRNA levels were dramatically increased at 6-24 hours postexposure. IL-1alpha mRNA levels were not increased following HD exposure. Immunohistochemical studies demonstrated that IL-1beta and IL-6 protein was produced at multiple sites within the ear, including epithelial cells, inflammatory cells, hair follicles, sebaceous glands, the dermal microvasculature, smooth muscle, and the dermal connective tissue. An increase in the intensity of staining for IL-1beta, and IL-6 was observed in localized areas at 6 hours and was evident in multiple areas at 24 hours. Positive staining for GM-CSF immunoreactive protein was localized to the inflammatory cells within the dermis. The number of immunostaining cells was increased as early as 1 hour following HD exposure. These studies document an early increase in the in vivo expression of inflammatory cytokines following cutaneous HD exposure. An understanding of the in vivo cytokine patterns following HD skin exposure may lead to defining the pathogenic mechanisms of HD injury and the development of pharmacological countermeasures.
皮肤接触化学战剂硫芥（双（2-氯乙基）硫醚，HD）会引发延迟性炎症皮肤反应和严重的组织损伤。尽管已知炎症细胞因子在应对皮肤损伤化学物质时产生或释放所起的作用，但与HD诱导的皮肤发病机制相关的体内细胞因子反应仍未得到充分了解。此外，关于暴露于HD后细胞因子基因表达的体内时间顺序的信息也很少。这些研究的目的是在HD攻击后24小时内确定HD皮肤损伤的体内分子生物标志物。通过定量逆转录-聚合酶链反应（RT-PCR）检测小鼠耳部水疱模型中白细胞介素1β（IL-1β）、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素6（IL-6）和白细胞介素1α（IL-1α）的基因表达。在3小时时首次观察到IL-1β mRNA水平升高；暴露后6至24小时，IL-1β、GM-CSF和IL-6 mRNA水平显著升高；HD暴露后IL-1α mRNA水平未升高。免疫组织化学研究表明，IL-1β和IL-6蛋白在耳部的多个部位产生，包括上皮细胞、炎症细胞、毛囊、皮脂腺、真皮微血管、平滑肌和真皮结缔组织。在6小时时，局部区域观察到IL-1β和IL-6染色强度增加，在24小时时在多个区域明显。GM-CSF免疫反应蛋白的阳性染色定位于真皮内的炎症细胞；HD暴露后1小时，免疫染色细胞数量就开始增加。这些研究证明了皮肤暴露于HD后体内炎症细胞因子表达的早期增加。了解HD皮肤暴露后的体内细胞因子模式可能有助于确定HD损伤的致病机制和开发药理学对策。
