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目的探讨烟曲霉菌孢子刺激对角膜上皮细胞中髓样细胞表达的激发受体-1(TREM-1)的表达及对炎症因子表达影响。方法在体外用烟曲霉菌孢子刺激角膜上皮细胞3 h、6 h和9 h,RT-PCR和Western blot检测细胞中TREM-1的表达。用TREM-1抑制剂LP17预处理角膜上皮细胞90 min后用烟曲霉菌孢子刺激,RT-PCR和Western blot检测细胞中TREM-1的表达,ELISA法检测细胞分泌的白介素6(IL-6)、白介素8(IL-8)水平,Western blot检测细胞中Toll样受体4(TLR4)、髓样分化因子(My D88)蛋白水平。结果烟曲霉菌孢子刺激后角膜上皮细胞中TREM-1 mRNA和蛋白表达水平均明显升高,并且随着刺激时间的增加而增多。LP17预处理能够有效降低烟曲霉菌孢子诱导的角膜上皮细胞中TREM-1 mRNA和蛋白的表达水平,并且对烟曲霉菌孢子诱导的角膜上皮细胞分泌的IL-6、IL-8和细胞中TLR-4、My D88蛋白表达具有拮抗作用。结论烟曲霉菌孢子刺激后角膜上皮细胞中TREM-1表达升高,抑制TREM-1能够显著降低烟曲霉菌孢子感染后角膜上皮细胞炎症因子的释放,这可能与My D88、TLR-4有关。
Objective To investigate the effect of Aspergillus fumigatus spores on the expression of TREM-1 and the expression of inflammatory cytokines in corneal epithelial cells. Methods The corneal epithelial cells were stimulated with Aspergillus fumigatus spores for 3 h, 6 h and 9 h, and the expression of TREM-1 was detected by RT-PCR and Western blot. Corneal epithelial cells were pretreated with TRAIL-1 inhibitor LP17 for 90 min and then stimulated with Aspergillus fumigatus spores. The expression of TREM-1 was detected by RT-PCR and Western blot. The levels of interleukin-6 (IL-6) , Interleukin 8 (IL-8) and Western blot were used to detect the protein levels of TLR4 and My D88. Results TREM-1 mRNA and protein expression in corneal epithelial cells were significantly increased after Aspergillus fumigatus spores stimulation, and increased with the stimulation time. LP17 pretreatment can effectively reduce the expression level of TREM-1 mRNA and protein in corneal epithelial cells induced by Aspergillus fumigatus spores and also inhibit the secretion of IL-6, IL-8 and TLRs in corneal epithelial cells induced by Aspergillus fumigatus spores -4, My D88 protein expression has antagonistic effect. Conclusion The expression of TREM-1 in corneal epithelial cells induced by Aspergillus fumigatus spores is up-regulated. Inhibition of TREM-1 can significantly reduce the release of inflammatory cytokines in corneal epithelial cells after Aspergillus fumigatus spores infection, which may be related to My D88 and TLR-4.