毛发是评估下丘脑-垂体-肾上腺轴(HPA轴)慢性活性变化很好的样品,毛发皮质醇的提取方法非常重要。为保证毛发皮质醇提取过程的可靠性,实验使用了液氮冷冻研磨;设计了空白对照组(磷酸缓冲液(PBS))(n=3)、原管组(n=10)和新管组(n=10),以检验缓冲液、离心管对提取的潜在干扰;对同一动物样品(n=16)两次独立提取的结果及毛发皮质醇浓度与受到的极端攻击量(血浆皮质醇浓度的间接指标)(n=16)进行了Pearson相关分析。结果表明:空白对照组测量结果为0pg/mg;新管浓度值与原管无显著性差异(F(1,19)=0.041,P=0.843,one-wayANOVA);两次独立提取结果高度正相关(r=0.893,P=0.000);皮质醇浓度与受到的极端攻击量中度正相关(r=0.591,P=-0.008)。因此,离心管、缓冲液对实验没有显著性干扰,溶液中皮质醇来源于毛发样品;提取操作流程可靠,所得数据可信。
Hair provides a suitable sample inassessing long-term changes in hypothalamic-pituitary-adrenocortical axis (HPA) system activity. The procedure used for hair analysis is therefore very important. To ensure that our procedure is reliable, liquid nitrogen was used to deepfreeze the samples before powdering them. Experimental trials used three treatment types: blank group (phosphate buffered saline (PBS)) (n=3), old tube group (n=10) and new tube group (n=10). The potential effects of PBS and tubes was then investigated. Additionally, both the relationship between the results of two samples of the same individuals (n= 16) yielded by the same procedures but at different times, and the relationship between hair cortisol concentrations and the number of received intense aggressions (n= 16) were also investigated. The results showed that the cortisol concentration of the blank group was 0pg/mg; new tubes did not differ from old tubes (F (1, 19) =0.041, P=0.843, one-way ANOVA); and there was a high association between the results of two samples of the same individuals, yielded by same procedures operated at different times (r=0.893, P=0.000). Moreover, hair cortisol concentrations were moderately correlated with the number of received intense aggressions (r=0.591, P=-0.008). Therefore, PBS and tubes did not affect the results significantly, therefore, our procedure was deemed reliable.