Patlolla Ram R, Mallampati Ramya, Fulzele Suniket V, Babu R Jayachandra, Singh Mandip
College of Pharmacy and Pharmaceutical Sciences, Florida A&M University, Tallahassee, FL 32307, USA.
Toxicol Lett. 2009 Mar 28;185(3):168-74. doi: 10.1016/j.toxlet.2008.12.012. Epub 2008 Dec 30.
In the present study we made an attempt to understand the skin irritation cascade of selected aliphatic hydrocarbons using microdialysis technique. Microdialysis probes were inserted into dermis in the dorsal skin of hairless rats. After 2h of probes insertion, occlusive dermal exposure (2h) was carried out with 230 microl of nonane, dodecane and tetradecane, using Hill top chambers((R)). Inflammatory biomarkers such as substance P (SP), alpha-melanocyte stimulating hormone (alpha-MSH) Interleukin 6 (IL-6) and prostaglandin E2 (PGE(2)) were analyzed in the dialysis samples by enzyme immunoassay (EIA). SP, alpha-MSH and IL6 were released in significant amounts following the dermal exposure of nonane and dodecane, whereas tetradecane did not induce any of these markers in significant amounts compared to control. Nonane increased the PGE(2) levels in significant amounts within 2h of chemical exposure compared to dodecane and tetradecane. IL-6 response was found to be slow and 2-3-fold increase in IL-6 levels was observed after 5h following nonane and dodecane application. The magnitude of skin irritation exerted by all three chemicals was in the order of nonane>or=dodecane>or=tetradecane. The results demonstrate that microdialysis can be used to measure the inflammatory biomarkers in the skin irritation studies and irritation response of chemicals was quantifiable by this method. In conclusion, microdialysis was found to be an excellent tool to measure several inflammatory biomarkers as a function of time after dermal exposures with irritant chemicals.
在本研究中,我们尝试使用微透析技术来了解所选脂肪族烃类的皮肤刺激级联反应。将微透析探针插入无毛大鼠背部皮肤的真皮层。插入探针2小时后,使用Hill top chambers((R)),用230微升壬烷、十二烷和十四烷进行2小时的封闭性皮肤暴露。通过酶免疫测定法(EIA)分析透析样本中的炎症生物标志物,如P物质(SP)、α-黑素细胞刺激素(α-MSH)、白细胞介素6(IL-6)和前列腺素E2(PGE(2))。壬烷和十二烷皮肤暴露后,SP、α-MSH和IL6大量释放,而与对照组相比,十四烷未大量诱导这些标志物中的任何一种。与十二烷和十四烷相比,壬烷在化学暴露后2小时内大量增加PGE(2)水平。发现IL-6反应缓慢,在施用壬烷和十二烷后5小时观察到IL-6水平增加2至3倍。所有三种化学物质引起的皮肤刺激程度顺序为壬烷≥十二烷≥十四烷。结果表明,微透析可用于皮肤刺激研究中测量炎症生物标志物,并且该方法可量化化学物质的刺激反应。总之,发现微透析是一种用于测量与刺激性化学物质皮肤暴露后随时间变化的多种炎症生物标志物的优秀工具。