牡荆素对小鼠烟曲霉菌性角膜炎的保护作用及其机制  

Effect of vitexin on Aspergillus fumigatus keratitis in mice and its mechanism

作　　者：徐海宁 易雯丹 贾奕奕 李翠[1] 尹敏[1] 顾凌雯 赵桂秋[1] XU Haining;YI Wendan;JIA Yiyi;LI Cui;YIN Min;GU Lingwen;ZHAO Guiqiu(Department of Ophthalmology,The Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院眼科,山东青岛266003 [2]威海卫人民医院眼科

出　　处：《精准医学杂志》2025年第1期13-18,共6页Journal of Precision Medicine

基　　金：国家自然科学基金面上项目(82171029)。

摘　　要：目的探究牡荆素对小鼠烟曲霉菌性角膜炎的保护作用及其机制。方法采用CCK-8实验检测牡荆素对HCECs及RAW264.7细胞细胞存活率的影响;将C57BL/6小鼠随机分为3组:对照组、烟曲霉菌性角膜炎组及牡荆素处理组(小鼠球结膜下注射100 mg/L的牡荆素10μL,每天1次,连续3 d),采用临床评分法评估小鼠真菌性角膜炎的严重程度;采用苏木精-伊红(HE)染色检测小鼠角膜炎症细胞浸润情况;采用平板计数实验检测小鼠角膜真菌载量;采用实时荧光定量PCR方法检测小鼠及RAW264.7细胞IL-1β、IL-6、MIP-2和LOX-1 mRNA水平的变化;采用蛋白免疫印迹实验检测RAW264.7细胞中LOX-1蛋白表达情况。结果牡荆素低于100 mg/L时对HCECs细胞活性无明显影响,牡荆素低于150 mg/L时对RAW264.7细胞活性无明显影响。与烟曲霉菌性角膜炎组小鼠相比,牡荆素处理组小鼠角膜炎病程进展较慢,小鼠角膜炎症细胞浸润水平较低,小鼠角膜真菌载量明显减少(t′=19.27,P<0.05);同时牡荆素处理组小鼠炎症因子IL-1β、IL-6、MIP-2和LOX-1 mRNA的表达水平与烟曲霉菌性角膜炎组小鼠相比均明显降低(t=4.95~6.15,P<0.05)。此外,牡荆素预处理后的RAW264.7细胞IL-1β、IL-6、MIP-2和LOX-1的mRNA及LOX-1蛋白水平与菌丝刺激组相比明显降低(t=-64.68~40.02,P<0.05)。结论牡荆素对烟曲霉菌刺激的RAW264.7细胞及真菌性角膜炎小鼠角膜具有保护作用,其机制可能是通过下调RAW264.7细胞以及小鼠角膜IL-1β、IL-6、MIP-2及LOX-1的mRNA和模式识别受体LOX-1蛋白表达而实现的。Objective To investigate the protective effect of vitexin against Aspergillus fumigatus(A.fumigatus)ke-ratitis in mice and its mechanism.Methods The effect of vitexin on the survival rate of human corneal epithelial cells(HCECs)and RAW264.7 cells was evaluated using Cell Counting Kit-8.Mice were divided into three groups:control group,A.fumigatus keratitis group,and vitexin-treated group(10μL of 100 mg/L vitexin was injected subconjunctivally once a day for 3 consecutive days).The severity of fungal keratitis(FK)was evaluated using clinical scores.Hematoxylin-eosin staining was used to detect the inflammatory cell infiltration in the mice.Plate counting was used to determine corneal fungal load in the mice.Changes in the mRNA expression levels of IL-1β,IL-6,MIP-2,and LOX-1 in the mice and RAW264.7 cells were measured using quantitative real-time PCR.The expression level of LOX-1 protein in RAW264.7 cells was determined using Western blotting.Results Vitexin did not have a significant effect on the viability of HCECs cells at a concentration lower than 100 mg/L and did not have a significant effect on the viability of RAW264.7 cells at a concentration lower than 150 mg/L.Compared with A.fumigatus keratitis group mice,vitexin-treated mice showed a slower course of keratitis progression,decreased inflammatory cell infiltration in the cornea,and a significantly reduced corneal fungal load(t′=19.27,P<0.05).Meanwhile,the mRNA expression levels of IL-1β,IL-6,MIP-2,and LOX-1 in the vitexin-treated group were significantly reduced compared with those in A.fumigatus keratitis group(t=4.95-6.15,P<0.05).The mRNA expression levels of IL-1β,IL-6,MIP-2,and LOX-1 and the protein expression level of LOX-1 were significantly reduced in vitexin-treated RAW264.7 cells compared with those in hyphae-stimulated cells(t=-64.68-40.02,P<0.05).Conclusion Vitexin has a protective effect on A.fumigatus-stimulated RAW264.7 cells and FK in mice cornea,which may be achieved by downregulating the mRNA expression of IL-1β,IL-6,MIP-2,and LO

关 键 词：真菌性角膜炎 烟曲霉菌 牡荆素 抗炎作用 LOX-1 

分 类 号：R772.21[医药卫生—眼科]