[1]刘晓瑞,杨淑芳,曾峥,等.利妥昔单抗对实验性自身免疫性葡萄膜炎小鼠的抗炎作用及其机制[J].眼科新进展,2023,43(6):434-438.[doi:10.13389/j.cnki.rao.2023.0087]
　LIU Xiaorui,YANG Shufang,ZENG Zheng,et al.Anti-inflammatory effect and mechanism of Rituximab on mice with experimental autoimmune uveitis[J].Recent Advances in Ophthalmology,2023,43(6):434-438.[doi:10.13389/j.cnki.rao.2023.0087]
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利妥昔单抗对实验性自身免疫性葡萄膜炎小鼠的抗炎作用及其机制/HTML
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《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
43卷
期数:
2023年6期
页码:
434-438
栏目:
实验研究
出版日期:
2023-06-05

文章信息/Info

Title:
Anti-inflammatory effect and mechanism of Rituximab on mice with experimental autoimmune uveitis
作者:
刘晓瑞; 杨淑芳; 曾峥; 丁然然; 赵宇; 由彩云
300052　天津市，天津医科大学总医院眼科（刘晓瑞，杨淑芳，丁然然，赵宇，由彩云）；300052　天津市，天津医科大学总医院神经外科（曾峥）
Author(s):
LIU Xiaorui1; YANG Shufang1; ZENG Zheng2; DING Ranran1; ZHAO Yu1; YOU Caiyun1
1.Department of Ophthalmology,Tianjin Medical University General Hospital,Tianjin 300052,China
2.Department of Neurosurgeon,Tianjin Medical University General Hospital,Tianjin 300052,China
关键词:
实验性自身免疫性葡萄膜炎; 利妥昔单抗; 白细胞介素-6; 白细胞介素-10
Keywords:
experimental autoimmune uveitis;  Rituximab;  interleukin-6;  interleukin-10
分类号:
R773
DOI:
10.13389/j.cnki.rao.2023.0087
文献标志码:
A
摘要:
目的　探讨利妥昔单抗（RTX）对实验性自身免疫性葡萄膜炎（EAU）小鼠的抗炎作用及其机制。
方法　取SPF级6～8周龄健康雌性C57BL/6J小鼠27只，使用光感受器间维生素A类结合蛋白1-20(IRBP1-20)建立EAU小鼠模型，将EAU小鼠随机分为PBS对照组、50 mg·kg-1 RTX实验组和100 mg·kg-1 RTX实验组，每组各9只小鼠，PBS对照组小鼠腹腔注射PBS,50 mg·kg-1 RTX实验组和100 mg·kg-1 RTX实验组小鼠每千克体重分别腹腔注射50 mg RTX和100 mg RTX。造模后第21天,对各组小鼠进行眼底图像的采集与视网膜组织HE切片的制作，通过小动物视网膜成像仪采集各组小鼠眼底图像并参照Caspi临床评分标准进行炎症评分,将HE染色切片置于显微镜下观察并进行拍照，参照Caspi组织病理学评分标准进行评分。采用实时荧光定量PCR检测各组小鼠视网膜组织白细胞介素（IL）-1β、IL-6、IL-4和IL-10的mRNA相对表达量。采用SPSS 21.0软件进行统计学分析。
结果　眼底图像显示：PBS对照组EAU小鼠眼底出现多发性血管炎，血管扩张，视网膜脉络膜出现多发病灶及少量线样病变；50 mg·kg-1 RTX实验组EAU小鼠眼底发生轻度血管炎，视网膜出现个别局部病灶，无线样病变；100 mg·kg-1RTX实验组EAU小鼠视网膜局部出现个别病灶，无线样病变。PBS对照组、50 mg·kg-1 RTX实验组和100 mg·kg-1RTX实验组EAU小鼠眼底图像Caspi临床评分分别为（1.944±0.808）分、（1.667±0.791）分、（1.667±0.829）分，三组间比较差异无统计学意义（P>0.05）。PBS对照组、50 mg·kg-1 RTX实验组和100 mg·kg-1RTX实验组EAU小鼠视网膜组织病理学评分分别为（1.563±1.116）分、（1.063±0.623）分、（1.250±0.655）分，三组间比较差异无统计学意义（P>0.05）。实时荧光定量PCR检测结果显示，50 mg·kg-1 RTX实验组和100 mg·kg-1 RTX实验组小鼠视网膜组织IL-6、IL-10 mRNA相对表达量以及IL-10/IL-6比值均高于PBS对照组（均为P<0.05）；100 mg·kg-1 RTX实验组小鼠视网膜组织IL-6、IL-10 mRNA相对表达量以及IL-10/IL-6比值均高于50 mg·kg-1 RTX实验组（均为P<0.05）。
结论　腹腔注射RTX可一定程度缓解EAU小鼠视网膜组织的炎性病变，其机制可能与RTX提高小鼠视网膜组织IL-10/IL-6比值有关。
Abstract:
Objective　To investigate the anti-inflammatory effect of Rituximab (RTX) on experimental autoimmune uveitis (EAU) mice and its mechanism.
Methods　A total of 27 healthy female C57BL/6J mice aged 6 to 8 weeks in specific pathogen-free grade were selected and the EAU mouse model was established using interphotoreceptor retinoid-binding protein 1-20 (IRBP1-20). EAU mice were randomly divided into the phosphate-buffered saline (PBS) control group, 50 mg·kg-1 RTX experimental group and 100 mg·kg-1 RTX experimental group, with 9 mice in each group. Mice in the PBS control group were intraperitoneally injected with PBS, while mice in the 50 mg·kg-1 RTX experimental group and 100 mg·kg-1 RTX experimental group were intraperitoneally injected with 50 mg RTX and 100 mg RTX per kg body weight, respectively. At 21 d after the modeling, fundus images of mice in each group were collected and hematoxylin-eosin (HE) staining slices of retinal tissue were prepared. Fundus images of mice in each group were collected by a small animal retinal imager and inflammation scores were obtained according to Caspi clinical scoring criteria. HE staining slices were observed and photographed under a microscope, and the score was made according to the Caspi histopathological scoring criteria. The relative messenger ribonucleic acid (mRNA) expressions of interleukin (IL)-1β, IL-6, IL-4 and IL-10 in the retinal tissues of mice in each group were detected by real-time quantitative PCR. SPSS 21.0 software was used for statistical analysis.
Results　Fundus images showed that the EAU mice in the PBS control group presented multiple vasculitis and vasodilatation in the fundus, as well as multiple lesions and a few linear lesions in the retinal choroid. The EAU mice in the 50 mg·kg-1 RTX experimental group showed mild vasculitis in the fundus, a few local lesions and no linear lesions in the retina. The EAU mice in the 100 mg·kg-1 RTX experimental group showed a few local retinal lesions and no linear lesions in the retina. The Caspi clinical scores in fundus images of EAU mice in the PBS control group, 50 mg·kg-1 RTX experimental group and 100 mg·kg-1 RTX experimental group were (1.944±0.808) points, (1.667±0.791) points and (1.667±0.829) points, with no significant difference (P>0.05). The retinal histopathological scores of EAU mice in the PBS control group, 50 mg·kg-1 RTX experimental group and 100 mg·kg-1 RTX experimental group were (1.563±1.116) points, (1.063±0.623) points and (1.250±0.655) points, with no significant difference (P>0.05). The results of real-time quantitative PCR showed that the relative mRNA expressions of IL-6 and IL-10 and the IL-10/IL-6 ratio in the retinal tissue of mice in the 50 mg·kg-1 RTX experimental group and 100 mg·kg-1 RTX experimental group were higher than those of the PBS control group (all P<0.05); the relative mRNA expressions of IL-6 and IL-10 and the IL-10/IL-6 ratio of mice in the 100 mg·kg-1 RTX experimental group were higher than those of the 50 mg·kg-1 RTX experimental group (all P<0.05).
Conclusion　Intraperitoneal injection of RTX can alleviate the inflammatory lesions in the retinal tissue of EAU mice to a certain extent, and the mechanism may be related to the increase of IL-10/IL-6 ratio in the retinal tissue.

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备注/Memo

备注/Memo:
国家自然科学基金项目（编号：81800817）
更新日期/Last Update: 2023-06-05