丁香酚通过PI3K/AKT信号通路减轻镰刀菌诱导的角膜炎症  

Eugenol attenuates the inflammation of Fusarium-induced keratitis through PI3K/AKT signaling pathway

作　　者：丁辉 胡施思 杨镇朵 陈晓丹 Ding Hui;Hu Shisi;Yang Zhenduo;Chen Xiaodan(Department of Myopia and Optometry,Hainan Eye Hospital Hainan Key Laboratory of Ophthalmology,Haikou 570311,Hainan Province,China)

机构地区：[1]海南省眼科医院近视屈光专科、海南省眼科学重点实验室,海南省海口市570311

出　　处：《国际眼科杂志》2024年第8期1194-1199,共6页International Eye Science

基　　金：海南省重点研发计划项目(No.ZDYF2021SHFZ232)。

摘　　要：目的:探讨丁香酚对镰刀菌(F.solani)诱导的小鼠真菌性角膜炎(FK)的保护作用,并初步探讨可能的潜在机制。方法:采用改良的表层镜法制备FK小鼠模型。二甲基亚砜(DMSO)组大鼠右眼结膜囊涂抹等量DMSO(0.05%)。丁香酚组将丁香酚(160μg/mL)涂抹至小鼠右眼结膜囊。类胰岛素生长因子-1(IGF-1)组除施用丁香酚外,还于右眼结膜囊涂抹PI3K/AKT通路激活剂IGF-1(10 nmol/mL)。分别在接种镰刀菌悬浮液的第1、3、5 d,于裂隙灯显微镜下观察角膜形态。采用苏木精伊红(HE)染色评估角膜组织病理损伤。测定角膜组织载菌量。酶联免疫吸附试验和蛋白免疫印迹用于分析炎症介质白介素-6(IL-6)和白介素-1β(IL-1β)的水平和PI3K/AKT通路蛋白的表达。结果:丁香酚治疗可改善FK小鼠的角膜炎形态症状和炎症反应,减轻角膜病理组织损伤和真菌负荷。在镰刀菌感染3 d,与DMSO组相比,丁香酚组角膜组织IL-6水平明显升高,而IL-1β水平明显降低(均P<0.05);且丁香酚组角膜组织IL-6水平明显高于IGF-1组、IL-1β水平明显低于IGF-1组(均P<0.05)。在感染5 d,丁香酚组角膜组织中IL-6和IL-1β水平均明显低于DMSO组和IGF-1组(P<0.05)。与DMSO组相比,丁香酚组角膜组织中p-PI3K和p-Akt表达均明显降低(P<0.05);且丁香酚组角膜组织p-PI3K和p-Akt表达明显低于IGF-1组(均P<0.05)。结论:丁香酚可能通过抑制PI3K/AKT通路来减轻镰刀菌引起的角膜炎症,对小鼠镰刀菌角膜炎具有保护作用。AIM:To investigate the protective effect of eugenol against Fusarium solani(F.solani)-induced fungal keratitis(FK)in mice and to preliminarily explore possible underlying mechanisms.METHODS:A modified epifluorescence microscopy method was used to prepare the FK mouse model.An equal amount of DMSO(0.05%)was applied to the conjunctiva of the right eye of rats in the dimethyl sulfoxide(DMSO)group.The eugenol group was prepared by applying eugenol(160μg/mL)to the conjunctival sac of the right eye of mice.The insulin-like growth factor-1(IGF-1)group was coated with the PI3K/AKT pathway activator IGF-1(10 nmol/mL)in the conjunctival sac of the right eye in addition to the administration of eugenol.The corneal morphology was observed under a slit-lamp microscope on days 1,3,and 5 of inoculation with F.solani suspension,respectively.Hematoxylin eosin(HE)staining was used to assess corneal histopathological damage.The bacterial load of corneal tissue was determined.Enzyme-linked immunosorbent assay and Western blot were used to analyze the levels of inflammatory mediators interleukin-6(IL-6)and interleukin-1β(IL-1β)and the expression of PI3K/AKT pathway proteins.RESULTS:Eugenol treatment improved the morphological symptoms of keratitis and inflammatory response in FK mice,and reduced corneal pathologic tissue damage and fungal load.At 3 d after F.solani infection,corneal tissue IL-6 levels were significantly higher and IL-1βlevels were significantly lower in the eugenol group compared with the DMSO group(both P<0.05);corneal tissue IL-6 levels were significantly higher and IL-1βlevels were significantly lower in the eugenol group than in the IGF-1 group(both P<0.05).At 5 d after infection,both IL-6 and IL-1βlevels in corneal tissue of the eugenol group were significantly lower than those of the DMSO and IGF-1 groups(P<0.05);compared with the DMSO group,the expression of p-PI3K and p-Akt in the corneal tissues of the eugenol group was significantly reduced(P<0.05);the expression of p-PI3K and p-Akt in corneal tissues

关 键 词：丁香酚 真菌性角膜炎 PI3K/AKT通路 炎症反应 真菌载量 

分 类 号：R772.21[医药卫生—眼科]