C57BL/6N小鼠早期视网膜变性及小胶质细胞活化状态研究  

Early retinal degeneration and activation of microglia in C57BL/6N mice

作　　者：孟欢 邓婷婷[3] 刘自强 侯小玉 马传政 苑维[3] 金明[3] Meng Huan;Deng Tingting;Liu Ziqiang;Hou Xiaoyu;Ma Chuanzheng;Yuan Wei;Jin Ming(Department of Ophthalmology,Beijing University of Chinese Medicine Third Affiliated Hospital,Beijing 100029,China;Beijing University of Chinese Medicine,Beijing 100029,China;Department of Ophthalmology,China-Japan Friendship Hospital,Beijing 100029,China;Eye Hospital,China Academy of Chinese Medical Sciences,Beijing 100040,China)

机构地区：[1]北京中医药大学第三附属医院眼科,北京市100029 [2]北京中医药大学,北京市100029 [3]中日友好医院眼科,北京市100029 [4]中国中医科学院眼科医院,北京市100040

出　　处：《国际眼科杂志》2024年第10期1536-1541,共6页International Eye Science

基　　金：中日友好医院院级科研基金项目(No.2019-2-MS-3)。

摘　　要：目的:观察C57BL/6N(Crb1 rd8/rd8)小鼠早期视网膜变性以及小胶质细胞的活化情况。方法:取雄性SPF级C57BL/6N小鼠15只、C57BL/6J小鼠15只,正常饲养。分别在入组时,入组4、8、12 wk,使用Micron-Ⅲ小动物视网膜影像系统进行双眼彩色眼底照相检查计算病变数量、病变面积。观察结束后处死小鼠,摘取右侧眼球制备视网膜组织切片,进行HE染色后光镜下观察视网膜组织形态;采用免疫组织化学染色分析两组小鼠视网膜CX3CR1的表达水平及位置。摘取左侧眼球分离视网膜,使用Western-Blot检测CD86、CD206的表达情况,使用电化学发光法测定视网膜中IL-1β、IL-6、TNF-α、IL-4、IL-10炎性因子含量水平。结果:眼底彩照结果显示在入组4、8、12 wk,C57BL/6N组眼底病变数量均较入组时显著增加,与C57BL/6J组同时间点变化量比较均有差异(均P<0.05);眼底病变面积变化量两组间入组12 wk有差异(P<0.05),各组内变化量比较均无差异(均P>0.05);视网膜组织HE染色示:C57BL/6N组视网膜结构异常,细胞排列疏松、紊乱,光感受器层向视网膜内侧明显的玻璃膜疣样凸起,C57BL/6J组视网膜结构清晰,细胞排列有序,无明显异常;免疫组化结果示:C57BL/6N组视网膜中CX3CR1高表达于神经节细胞层、内外丛状层、感光细胞层及病灶处位置,平均光密度为0.285±0.056,C57BL/6J组为0.189±0.084(P<0.05);Western-Blot结果显示:与C57BL/6J组相比,C57BL/6N组视网膜CD86、CD206蛋白有不同程度升高,两组CD86蛋白表达有差异(P<0.05);细胞因子检测结果显示:C57BL/6N组IL-1β、TNF-α水平显著高于C57BL/6J组,而IL-10含量则明显较低(均P<0.05)。结论:C57BL/6N(Crb1 rd8/rd8)小鼠视网膜变性进展缓慢,随年龄呈进行性加重,病灶部位视网膜结构紊乱,伴有以M1型极化为主的小胶质细胞浸润。AIM:To observe the early retinal degeneration and activation of microglia in C57BL/6N(Crb1 rd8/rd8)mice.METHODS:Totally 15 male SPF C57BL/6N mice and 15 male SPF C57BL/6J mice were raised normally,and fundus photography examinations were performed by Micron-Ⅲat the time of 0,4,8,12 wk of enrollment to calculate the number and area of retinopathy.At the end of experiment,all mice were sacrificed and the right eyeballs were removed to prepare retinal tissue slices.After HE staining,the retinal tissue morphology was observed under optical microscope while the location and level of CX3CR1 expression were detected in immunohistochemical staining.The left eyeballs were removed to isolate retina,then Western-Blot was used to analyze the expression of CD86 and CD206 proteins in retina,and the concentration of IL-1β,IL-6,TNF-α,IL-4 and IL-10 in retina was detected by electrochemiluminescence.RESULTS:The result of fundus photography examinations showed that the number of retinopathy in the C57BL/6N significantly increased at 4,8,and 12 wk,and there were differences in variations compared with the C57BL/6J at the same time point(all P<0.05).In the changes in area of retinopathy,there was a difference between two groups at 12 wk(P<0.05),but no difference in variations within groups(both P>0.05).HE staining of retinal tissue showed that the retinal structure of C57BL/6N mice was abnormal,with loose and disordered cell arrangement,and the photoreceptor layer was obviously protruding to the inner side of retina with a drusen-like protrusion.The retinal structure of C57BL/6J mice was clearer,with orderly cell arrangement and no obvious abnormality.Immunohistochemical results showed that CX3CR1 was highly expressed in ganglion cell layer,inner and outer plexiform layer,photoreceptor cell layer and lesion in the retina of C57BL/6N mice,with a mean density of 0.285±0.056 in C57BL/6N and 0.189±0.084 in C57BL/6J mice(P<0.05).The results of Western-Blot showed that the expression of CD86 and CD206 in retina of C57BL/6N increased

关 键 词：C57BL/6N 视网膜变性 年龄相关性黄斑变性 小胶质细胞 极化 

分 类 号：R774.13[医药卫生—眼科]