烟曲霉对人THP-1细胞中IL-6分泌和信号分子IκBα活化的影响

Effect of Aspergillus fumigatus on the production of interleukin-6 and activation of IκBα in human THP-1 monocytes

目的:明确烟曲霉对THP-1细胞中IL-6水平和细胞内信号分子NF-κB抑制蛋白(IκBα)激活的影响。方法:体外培养THP-1细胞,分为10~6CFU/mL和10~7CFU/mL烟曲霉组,β-Glucan(100g/L)阳性对照组和空白对照组,RT-PCR和ELISA检测各组IL-6 mRNA和蛋白表达水平,免疫印迹法检测IκBα和磷酸化IκBα水平。结果:10~6 CFU/mL IL-6 mRNA水平低于10~7CFU/mL烟曲霉组,选择10~7CFU/mL烟曲霉组作为实验浓度。10~7CFU/mL烟曲霉组IL-6 mRNA和蛋白浓度分别为(209.38±25.35)ng/L和(879.86±32.35)ng/L高于空白对照组的(1.19±0.36)和(10.67±0.17)ng/L,差异均有统计学意义(均P<0.05)。烟曲霉作用于THP-1细胞后磷酸化IκBα蛋白水平显著升高。NF-κB信号通路抑制剂BAY 11-7082可显著降低烟曲霉组IL-6 mRNA的表达。结论:烟曲霉可能通过激活THP-1细胞中的IκBα刺激IL-6分泌。

Objective： To determine the effect of Aspergillus fumigatus on the production of interleukin-6 and activation of IκBα in an acute monocytic leukemia cell line THP-1. Methods： THP-l cells were cultured in vitro and divided into 10^6 CFU/mL and 10^7 CFU/mL A. fumigatus group,100 μg/mL β-Glucan control group and blank control group. The level of IL-6 mRNA and protein was detected by RT-PCR and ELISA respectively. The level of IκBα and phosphorylated IκBα was detected by Western blot. Results： The experimental concentration was selected to be 10^7 CFU/mL,because the level of IL-6 mRNA in the 10^6 CFU/mL A.fumigatus group was lower than that in the 10^7 CFU/mL group. The levels of IL-6 mRNA and protein in the 10^7 CFU/mL A. fumigatus group were 209.38±25.35 and 879.86±32.35 ng/L,which were higher than that in the blank control group（ 1.19±0.36 and 10.67±0.17 ng/L）,with significant differences（ Ps〈0.05）. The level of phosphorylated IκBα protein was increased significantly when induced by 10^7 CFU/mL A. fumigatus and was decreased when induced by NF-κB signaling pathway inhibitor of bay 11-7082. Conclusion： A. fumigatus stimulates the secretion of IL-6 in human THP-1 cells through activating IκBα.