烟曲霉菌孢子刺激对角膜上皮细胞中TREM-1的表达及炎症因子表达的影响

摘要

Objective To investigate the expression of TREM-1 in corneal epithelial cells stimulated by Aspergillus fumigatus spores and its effect on the expression of inflammatory cytokines.Methods In vitro stimulation of 3h, 6h and 9h in corneal epithelial cells with aspergillus fumigatus spores, detection of TREM-1 expression in cells by RT-PCR and Western blot.TREM-1 inhibitor LP17 was used to treat corneal epithelial cells after 90min stimulated by aspergillus fumigatus spores, detection of TREM-1 expression in cells by RT-PCR and Western blot, ELISA method was used to detect the levels of IL-6 and IL-8 secreted by the cells, detection of TLR-4 and MyD88 protein levels in cells by Western blot.Results The expression of TREM-1 mRNA and protein in corneal epithelial cells were significantly increased after aspergillus fumigatus spores stimulation, and the expression increased with the increase of stimulation time.LP17 pretreatment can effectively reduce the expression of TREM-1 mRNA and protein in corneal epithelial cells induced by Aspergillus fumigatus spores, and the expression of IL-6, IL-8 and TLR-4 and MyD88 protein in the epithelial cells induced by aspergillus fumigatus spores were antagonistic.Conclusion Increased expression of TREM-1 in corneal epithelial cells stimulated by Aspergillus fumigatus spores, inhibition of TREM-1 could significantly reduce the release of inflammatory cytokines in corneal epithelial cells infected by aspergillus fumigatus spores, this may be related to MyD88, TLR-4.%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(MyD88)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、MyD88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MyD88、TLR-4有关.