黄芪甲苷干预碘酸钠诱导的光感受器退行性病变的效应及相关机制  

Effects and mechanisms of astragaloside A treatment on sodium iodate-induced photoreceptor degeneration

作　　者：李梅 常捷 吴晗晗 徐静[1,2] 杜霄烨[1,2] 崔金刚 张腾[1,2] 陈瑜[1,2,3] Li Mei;Chang Jie;Wu Hanhan;Xu Jing;Du Xiaoye;Cui Jingang;Zhang Teng;Chen Yu(Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China;Clinical Research Institute of Integrative Medicine,Shanghai Academy of Traditional Chinese Medicine,Shanghai 200437,China;Laboratory of Clinical and Molecular Pharmacology,Yueyang Hospital of Integrated Traditional Chinese and Western Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai 200437,China)

机构地区：[1]上海中医药大学附属岳阳中西医结合医院,上海200437 [2]上海市中医药研究院中西医结合临床研究所,上海200437 [3]上海中医药大学附属岳阳中西医结合医院临床分子药理实验室,上海200437

出　　处：《中华眼底病杂志》2024年第6期454-462,共9页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金(面上项目)(82274264)。

摘　　要：目的观察并初步探讨黄芪甲苷(AS-A)干预碘酸钠(NaIO_(3))诱导的光感受器退行性病变的效应及相关机制。方法6~8周龄健康雄性C57BL/6J小鼠60只,随机分为正常对照组(NC组)、NaIO_(3)组、AS-A组,每组20只。建模前30 min,AS-A组按100 mg/kg的剂量腹腔注射100μl AS-A;30 min后,NaIO_(3)组、AS-A组小鼠按30 mg/kg的剂量腹腔注射100μl NaIO_(3)。其后,AS-A组小鼠每天早晚间隔12 h给药2次,直到实验结束。建模后1 d,紧密连接蛋白(ZO-1)染色观察视网膜色素上皮(RPE)细胞结构;实时荧光定量聚合酶链反应(qPCR)检测各组小鼠视网膜趋化因子配体2(Ccl2)、白细胞介素1β(Il-1β)、混合谱系激酶结构域样蛋白(Mlkl)、受体相互作用蛋白激酶3(Ripk3)、肿瘤坏死因子(Tnf)等基因的mRNA相对表达量。建模后3 d,免疫组织化学染色观察各组小鼠视网膜中离子钙接头蛋白1(Iba1)、神经胶质纤维酸性蛋白(GFAP)阳性表达;原位末端标记(TUNEL)法检测各组小鼠视网膜光感受器细胞的死亡情况。建模后4 d,采用眼底彩色照相、光相干断层扫描(OCT)检查观察各组小鼠眼底形态;苏木精-伊红染色(HE)观察各组小鼠视网膜形态结构。两组间比较采用独立样本t检验;三组间比较采用单因素方差分析。结果眼底彩色照相、OCT检查发现,NaIO_(3)组小鼠眼底可见大量散在黄白色玻璃膜疣样结构,RPE层出现大量强反射区;AS-A组RPE层中强反射区数量减少。ZO-1染色结果显示,NaIO_(3)组RPE细胞膜上ZO-1大量丢失;AS-A组ZO-1均匀分布于RPE细胞膜上。HE染色结果显示,NaIO_(3)组RPE层可见圆形黑色沉积物,光感受器内外节结构严重受损,外核层(ONL)细胞层核数显著减少;AS-A组RPE层色素整齐,光感受器内外节结构完整,ONL细胞核层数显著增加。TUNEL检测结果显示,NaIO_(3)组ONL可见大量TUNEL阳性细胞核,AS-A组视网膜ONL中TUNEL阳性细胞核数量显著减少,差异有统计学意义(t=2.66,P<0.05)。Objective To investigate the effect of astragaloside A(AS-A)on the photoreceptor degeneration induced by sodium iodate(NaIO_(3))and its related mechanism.Methods Sixty healthy male C57BL/6J mice,aged 6-8 weeks,were randomly divided into normal control(NC)group,NaIO_(3)group,and ASA group,with twenty mice in each group.30 min before modeling,AS-A group mice were intraperitoneally injected with 100μl AS-A at a dose of 100 mg/kg body weight.30 min later,mice in NaIO_(3)group and AS-A group were intraperitoneally injected with 100μl NaIO_(3)at a dose of 30 mg/kg body weight.Subsequently,AS-A group mice were administered AS-A twice daily at 12 h intervals until the end of the experiment.On day 1 postmodeling,zonula occludens-1(ZO-1)immunohistochemistry was performed to observe the structure of retinal pigment epithelium(RPE)cells;real-time quantitative polymerase chain reaction(qPCR)was conducted to detect the mRNA expression of various retinal chemokine ligand-2(Ccl2),interleukin-1 beta(Il-1β),mixed lineage kinase domain-like protein(Mlkl),receptor-interacting protein kinase 3(Ripk3),and tumor necrosis factor(Tnf).On day 3 post-modeling,immunohistochemistry was performed to observe the expression of ionized calcium binding adaptor molecule 1(Iba1)and glial fibrillary acid protein(GFAP)in the retina;TdT-mediated dUTP nick-end labeling(TUNEL)assay was used to detect photoreceptor cell death in each group.On day 4 post-modeling,fundus morphology of mice in each group was observed by fundus color photography and optical coherence tomography(OCT).Hematoxylin-eosin staining(HE)was used to observe the morphological structure of the retina in each group.Inter-group comparisons between two groups were conducted using independent samples t-test,while comparisons among three groups were performed using one-way ANOVA.Results Fundus color photography and OCT examination showed that a large number of scattered yellowwhite subretinal nodular structures in the fundus of NaIO_(3)group mice,and a large number of strong reflection a

关 键 词：黄芪甲苷 视网膜 光感受器退行性病变 细胞死亡 小胶质细胞激活 Müller细胞反应性胶质增生 动物实验 

分 类 号：R774.1[医药卫生—眼科]