摘要
目的探讨扁柏酚(HK)对小鼠烟曲霉菌性角膜炎的保护作用及其机制。方法建立烟曲霉菌性角膜炎小鼠模型,于感染后第1天开始分别给予两组小鼠角膜10 mg/L HK或1 g/L DMSO点眼处理,于感染后第1天和第3天裂隙灯下观察小鼠角膜炎症情况;采用菌落形成单位计数法和过碘酸希夫染色法检测小鼠角膜真菌残存量;采用免疫荧光染色法检测小鼠角膜巨噬细胞数量的变化;以8、16 mg/L HK或1 g/L DMSO预处理RAW264.7巨噬细胞2 h,以烟曲霉菌菌丝刺激细胞8 h后,采用RT-qPCR法检测各组细胞IL-1β、单核细胞趋化蛋白-1(MCP-1)、趋化因子配体-1(CXCL-1)和Dectin-1 mRNA的相对表达水平。结果在感染后第3天,与对照组相比,HK组小鼠角膜炎症反应明显减轻,真菌残存量显著降低(F=366.23,P<0.05),巨噬细胞浸润减少。与1 g/L DMSO预处理组相比,8、16 mg/L HK预处理组可以显著降低烟曲霉菌诱导的RAW264.7巨噬细胞中炎症相关因子IL-1β、MCP-1、CXCL-1和模式识别受体Dectin-1 mRNA的表达(F=90.80~370.57,P<0.05)。结论HK可通过降低角膜真菌负荷,抑制角膜组织巨噬细胞募集,降低巨噬细胞炎症因子、趋化因子和Dectin-1 mRNA的表达,在烟曲霉菌性角膜炎中发挥保护作用。
Objective To investigate the protective effect of hinokitiol(HK)against Aspergillus fumigatus keratitis and related mechanism.Methods A mouse model of Aspergillus fumigatus keratitis was established,and the cornea was treated with the eye droppings of 10 mg/L HK or 1 g/L DMSO since day 1 after infection.The inflammatory changes of the cornea were observed under a slit lamp on days 1 and 3 after infection;colony-forming unit counting and periodic acid-Schiff staining were used to measure fungal residue in the cornea;immunofluorescent staining was used to measure the change in the number of macrophages in the cornea.After RAW264.7 macrophages were pretreated with 8 and 16 mg/L HK or 1 g/L DMSO for 2 hours and were stimulated with Aspergillus fumigatus hyphae for 8 hours,RT-qPCR was used to measure the relative mRNA expression levels of interleukin-1β(IL-1β),monocyte chemotactic protein-1(MCP-1),chemokine(C-X-C motif)ligand 1(CXCL-1),and Dectin-1.[WTHX]Results On day 3 after infection,compared with the control group,the HK group had significant alleviation of the inflammatory response of the cornea and significant reductions in fungal residue(F=366.23,P<0.05)and macrophage infiltration.Compared with the 1 g/L DMSO pretreatment group,the 8 and 16 mg/L HK pretreatment group had significant reductions in the mRNA expression of the inflammatory factors IL-1β,MCP-1,and CXCL-1 and the pattern recognition receptor Dectin-1 in RAW264.7 macrophages induced by Aspergillus fumigatus(F=90.80-370.57,P<0.05).Conclusion HK exerts a protective effect against Aspergillus fumigatus keratitis by reducing fungal load in the cornea,inhibiting macrophage recruitment in corneal tissue,and reducing the mRNA expression of inflammatory factors,chemokines,and Dectin-1 in macrophages.
作者
马淑芹
王谦
徐强
贺梦婷
林静
MA Shuqin;WANG Qian;XU Qiang;HE Mengting;LIN Jing(Department of Ophthalmology,The Affiliated Hospital of Qingdao University,Qingdao 266003,China)
出处
《精准医学杂志》
2022年第6期490-494,498,共6页
Journal of Precision Medicine
基金
山东省泰山学者青年专家人才项目(tsqn2018-12151)。
