【摘要】 目的研究真菌性角膜炎小鼠在不同光照条件下中性粒细胞的反应。方法取93只6-8周雄性SPF级C57BL/6小鼠,饲养在以下光照条件下,LD组小鼠从7:00-19:00接受光照(光强度110-210IW/cm~2),LL组接受24h光照(光强度110-210IW/cm~2),DD组24h黑暗,所有小鼠在LD环境下饲养7d,应用随机数字法,将小鼠平均分为三组,LD组、LL组、DD组,在相应光照条件下饲养12-15d。制作真菌性角膜炎模型,其中12只小鼠只用于临床观察,分别于造模前0h,造模后12h、18h、24h、36h、48h、72h、120h、168h,使用裂隙灯显微镜进行眼前段照相及临床评分。其余81只小鼠分别于以上时间点处死后进行取材,行免疫荧光染色角膜整铺片,使用单光子激光共聚焦显微镜,在40x油镜下对角膜整铺片进行扫描,使用图像处理软件,计算出不同时间点及不同实验条件处理下的小鼠角膜中中性粒细胞的浸润量。结果 LD组小鼠总体角膜炎症评分高于DD组,在造模后24h,LD组小鼠角膜炎症临床评分明显高于DD组,差异有统计学意义(P=0.008)。LL组小鼠总体角膜炎症评分高于DD组,在造模后18h、24h、36h、168h,LL组小鼠角膜炎症临床评分均明显高于DD组,差异有统计学意义(P=0.013,0.004,0.008,0.022<0.05)。LD组小鼠角膜中中性粒细胞浸润量总体高于DD组,在造模后12h、48h,LD组小鼠角膜中中性粒细胞的浸润量明显高于DD组,差异有统计学意义(P=0.033,0.009 <0.05)。LL组小鼠角膜中中性粒细胞浸润量总体高于DD组,在造模后12h、24h、48h,LL组小鼠角膜中中性粒细胞的浸润量明显高于DD组,差异有统计学意义(P=0.002,0.022,0.001 <0.05)。造模后三个组中性粒细胞与临床评分呈明显正相关(r=0.192、0.454,P=0.019、0.000)。结论在小鼠真菌感染阶段光有助于提高先天免疫细胞的招募,引起更多的中性粒细胞浸润,导致小鼠真菌性角膜炎更严重的炎症反应。更多还原

【Abstract】 Objective To study the response of neutrophils of fungal keratitis in mice under different light conditions.Methods 93 male SPF C57 BL/6 mice aged 6-8 w were selected and bred under the following light conditions,LD group mice received light from 7:00 to 19:00(light intensity 110-210 IW/cm~2),LL group received 24 h light(light intensity 110-210 IW/cm~2),DD group received 24 h dark,all mice were bred under LD environment for 7 days,the mice were divided into three groups, LD group, LL group and DD group,the mice were fed for 12-15 days under the corresponding light conditions. To make fungal keratitis model, among which 12 mice were only used for clinical observation, respectively 0 h before modeling, 12 h,18 h,24 h,36 h,48 h,72 h,120 h,168 h after modeling,and slit lamp microscope was used for anterior segment photography and clinical scoring.The rest of the 81 mice was performed after the above time point executed based respectively,immunofluorescence staining was performed on the whole cornea,using a single photon laser confocal microscope,the whole corneal film was scanned under 40 x oil lens,the infiltration of neutrophils in cornea of mice treated at different time points and under different experimental conditions was calculated by using image processing software.Results The total keratitis score of LD group was higher than that of DD group,at 24 h after modeling, the clinical score of corneal inflammation of LD group was significantly higher than that of DD group(P=0.008).The total keratitis score of LL group was higher than that of DD group,at 18 h, 24 h, 36 h and 168 h after modeling,the clinical scores of corneal inflammation of LL group were significantly higher than those of DD group(P=0.013, 0.004, 0.008, 0.022 < 0.05),at 12 h and 48 h after modeling, the infiltration of neutrophils in the cornea of LD group was significantly higher than that of DD group(P=0.033, 0.009 < 0.05).The infiltration of neutrophils in the cornea of LL group was higher than that of DD group,at 12 h, 24 h and 48 h after modeling, the infiltration of neutrophils in the cornea of LL group was significantly higher than that of DD group(P=0.002, 0.022, 0.001 < 0.05).The infiltration of neutrophils in the three groups was positively correlated with clinical scores(r = 0.192, 0.454, P = 0.019, 0.000).Conclusion In the stage of fungal infection in mice, light helps to increase the recruitment of neutrophils,more neutrophils infiltrated, leading to more severe inflammation of fungal keratitis in mice.更多还原