依达拉奉对视网膜脱离模型大鼠早期感光细胞自噬的调控作用  

Regulatory effect of edaravone on the photoreceptor autophagy at the early stage of experimental retinal detachment in rats

作　　者：康紫薇 任秀瑜 张妍春 裴澄[1] 张娜娜[3] 夏益敏 杨菁茹 Kang Ziwei;Ren Xiuyu;Zhang Yanchun;Pei Cheng;Zhang Nana;Xia Yimin;Yang Jingru(First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710049,China;Shaanxi Eye Hospital,Xi'an People's Hospital(Xi'an Fourth Hospital),Xi'an Ocular Fundus Disease Research Institute,Affiliated Guangren Hospital,School of Medicine,Xi'an Jiaotong University,Xi'an,710004,China;Cancer Research Institute of Basic Medical Science School,Xi'an Jiaotong University,Xi'an 710061,China;Pathology Department of Xi'an People's Hospital(Xi'an Fourth Hospital),Xi'an 710004,China)

机构地区：[1]西安交通大学第一附属医院,710049 [2]西安市人民医院(西安市第四医院),陕西省眼科医院,西安市眼底病研究所,西安交通大学医学院附属广仁医院眼科,710004 [3]西安交通大学癌症研究所,710061 [4]西安市人民医院(西安市第四医院)病理科,710004

出　　处：《中华实验眼科杂志》2021年第9期776-784,共9页Chinese Journal Of Experimental Ophthalmology

基　　金：陕西省自然科学基础研究计划面上项目(2015JM8408);陕西省中医管理局中医药科研课题面上项目(JCMS063);陕西省重点研发计划项目(2021SF-162);西安市科技计划重大研究项目[201805104YX12SF38(3)];西安市第四医院科研孵化基金项目(FZ-26)。

摘　　要：目的研究氧自由基清除剂依达拉奉对视网膜脱离(RD)模型大鼠早期视网膜自噬的调控及感光细胞的保护作用。方法取51只成年雄性Sprague-Dawley大鼠进行RD造模,另取24只大鼠作为PBS注射组。造模组采用右眼视网膜下注射0.5%透明质酸钠的方法制作RD动物模型。将造模成功大鼠按照随机数字表法随机分为RD模型组和依达拉奉治疗组。依达拉奉治疗组造模后连续给予3 mg/kg依达拉奉腹腔内注射,每天2次,PBS注射组和RD模型组腹腔注射等容量0.9%氯化钠溶液。分别于造模后1、3、7 d处死动物并取材。收集眼内液并检测总超氧化物歧化酶(T-SOD)活性及丙二醛(MDA)含量。Western blot法检测视网膜组织中超氧化物歧化酶2(SOD2)、核因子E2相关因子2(Nrf2)、自噬相关基因4(Atg4)、微管相关蛋白质1轻链3(LC3)蛋白表达。全眼球石蜡切片TUNEL染色分析感光细胞凋亡情况。结果所有RD模型眼视网膜脱离范围均>60%。不同时间点各组眼内液中MDA含量和T-SOD活性总体比较,差异均有统计学意义(MDA:F分组=385.513,P<0.01;F时间=13.021,P<0.01.T-SOD:F分组=48.865,P<0.01;F时间=7.700,P=0.003);与PBS注射组相比,造模后1、3、7 d RD模型组和依达拉奉治疗组眼内液MDA含量显著升高,T-SOD活性显著降低,差异均有统计学意义(均P<0.05);与RD模型组比较,造模后1、3、7 d依达拉奉治疗组MDA含量显著降低,T-SOD活性显著升高,差异均有统计学意义(均P<0.05)。与PBS注射组相比,RD模型组和依达拉奉治疗组造模后1、3、7 d视网膜中SOD2和Nrf2蛋白相对表达量显著升高,造模后1、3 d视网膜中Atg4、LC3B-Ⅱ/LC3B-Ⅰ相对表达量显著升高,差异均有统计学意义(均P<0.05);与RD模型组比较,依达拉奉治疗组造模后1、3、7 d视网膜中SOD2相对表达量显著升高,造模后1、3 d视网膜中Nrf2蛋白相对表达量显著升高,造模后3 d视网膜中Atg4、LC3B-Ⅱ/LC3B-Ⅰ蛋白相对表达量显著升高,�Objective To investigate the effect of edaravone,a free radical scavenger,on the regulation of retinal autophagy and the protection of photoreceptor cells at the early stage of experimental retinal detachment(RD)in rats.Methods Fifty-one adult male Sprague-Dawley rats were used for RD model establishment,and another 24 rats were served as PBS injection group.The RD model was established via subretinal injection of 0.5%sodium hyaluronate into the right eye of the rats and the rats with successful modeling were randomly divided into RD model group and edaravone treatment group.The rats in the edaravone treatment group were given edaravone of 3 mg/kg intraperitoneally,twice a day after modeling,and the rats in the PBS injection group and RD model group were given equal volume of normal saline.Rats were sacrificed on the 1st day,3rd day and 7th day following modeling.The T-superoxide dismutase(T-SOD)activity and malondialdehyde(MDA)content in the intraocular fluid was detected.The expression levels of superoxide dismutase 2(SOD2),nuclear factor E2-related factor 2(Nrf2),autophagy related gene 4(Atg4),microtubule-associated protein 1 light chain 3B(LC3B)and other proteins in retinal tissue were identified by Western blot analysis.TUNEL staining was performed on paraffin sections of the whole eyeball to analyze the apoptosis of photoreceptor cells.The study protocol was approved by an Ethics Committee of Xi'an Fourth Hospital(No.2016016).The use and care of animals complied with the Regulations on the Administration of Experimental Animals.Results The RD area was more than 60%in rat eyes of RD model.There were significant differences in MDA content and T-SOD activity among different groups at various time points(MDA:F group=385.513,P<0.01;F time=13.021,P<0.01.T-SOD:F group=48.865,P<0.01;F time=7.700,P=0.003).Compared with the PBS injection group,the MDA concentration was significantly increased and the T-SOD activity was significantly decreased in the RD group and edaravone treatment group on the 1st,3rd and 7th day af

关 键 词：视网膜脱离 依达拉奉 自噬 感光细胞凋亡 

分 类 号：R774.1[医药卫生—眼科]