范友羊,刘俊,周海燕,曾家顺,李龙
FAN Youyang,LIU Jun,ZHOU Haiyan,ZENG Jiashun,LI Long

• 1:贵州医科大学附属医院肾病风湿科
• 2:贵州医科大学
• 3:贵州医科大学附属医院临床医学研究中心

摘要(Abstract)：

目的:观察糖原合成酶激酶-3β(GSK-3β)抑制剂对胶原诱导的关节炎(CIA)小鼠的治疗作用及其与树突状细胞(DC)亚群变化的相关性。方法:取6～8周龄近交系雄性DBA/l小鼠19只,分为正常组(n=5)、模型组(n=7)及治疗组(n=7),模型组与治疗组小鼠制作CIA模型,治疗组小鼠于造模后第22天开始每天予GSK-3β抑制剂(TDZD-8) 5 mg/kg,模型组与正常组均予等量生理盐水,连续注射2周;观察和比较各组小鼠足趾肿胀程度及关节炎指数(AI)评分;第46天脱颈处死各组小鼠后切取右后肢踝关节,采用苏木素-伊红(HE)染色观察各组小鼠关节的组织学特征;采用免疫组织化学检测各组小鼠关节滑膜组织GSK-3β表达,采用流式细胞术检测各组小鼠脾脏树突状细胞(DC)中淋巴组织驻留型DC(cDC)及其CD8~+cDC、CD4~+CD8~-cDC、CD4~-CD8~-cDC各亚群变化。结果:与模型组比较,治疗组小鼠踝关节处红肿、出血点等现象明显好转;与模型组相比,治疗组小鼠AI评分在初次免疫后第33天开始下降,差异有统计学意义(P <0. 05或P <0. 01); HE染色结果显示为关节腔炎症状态明显改善;治疗组小鼠踝关节GSK-3β蛋白表达低于模型组,高于正常组,差异均有统计学意义(P <0. 05);治疗组小鼠脾脏总cDC、CD4~+cDC比例高于模型组,低于正常组,差异均有统计学意义(P <0. 05);治疗组小鼠脾脏CD4~-CD8c~-cDC比例低于模型组,高于正常组,差异均有统计学意义(P <0. 05)。结论:GSK-3β抑制剂可通过调节CIA小鼠脾脏cDC数量及各亚群的比例,缓解其关节病变及其病理表现。
Objective: To observe the therapeutic effect of glycogen synthase kinase-3β( GSK-3β inhibitor on collagen-induced arthritis( CIA) and its correlation with the changes of dendritic cell subsets. Methods: The male DBA/l mice of 6-8 week old were divided into the normal group( n =5),the model group( n = 7) and GSK-3β treatment group( n = 7). CIA models with reference to classical CIA model method for the model group and the treatment group were prepared. Then modeled mice were given 5 mg/kg TDZD-8 of reagents every day on the 22 ndday. The model group and the normal group were given the same amount of normal saline for 2 weeks. The scores of signs,toe swelling and arthritis index( AI) were observed and compared among the groups. On the 46 thday,the right hindlimb ankle joint was removed and the histological features of the joints were observed by hematoxylin-eosin( HE) staining,and the expression of synovial tissue GSK-3β was detected by immunohistochemistry,and the changes of lymphoid tissue retention DC( cDC) and its CD8~+cDC,CD4~+CD8~-cDC,CD4~-CD8~-cDC subsets in splenic dendritic cells were detected by flow cytometry.Results: Compared with the model group,GSK-3β treatment group mice diet,hair gloss,hair removal and joint redness,bleeding points and other phenomena were improved significantly. The AI scores of mice in the treatment group decreased more significantly on the 33 rdday after initial immunization than those in the model group,and the difference was statistically significant( P < 0. 05 or P < 0. 01). HE staining showed that the inflammatory state of articular cavity improved significantly. The expression of GSK-3β protein in the ankle joint in the treatment group was lower than that in the model group and higher than that in the normal group,and the difference was statistically significant( P < 0. 05). The proportion of total cDC,CD4~+cDC of spleen in the treatment group was higher than that in the model group and lower than that in the normal group,and the difference was statistically significant( P <0. 05). The proportion of spleen CD4~-CD8c~-cDC in the treatment group was lower than that in the model group and higher than that in the normal group,and the difference was statistically significant( P < 0. 05). Conclusion: GSK-3β inhibitors can relieve joint lesions and pathological manifestations by regulating the number of cDC spleen and the proportion of each subgroup in CIA mice.

关键词(KeyWords)： 关节炎,类风湿;小鼠;糖原合成酶激酶-3β;树突状细胞;脾脏;诱导型关节炎小鼠
arthritis,rheumatoid;mice;glycogen synthase kinase-3β(GSK-3β);dendritic cells;spleen;collagen-induced arthritis(CIA)

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81460254);; 贵州省科技计划项目[黔科合基础(2018)1137];; 贵州医科大学2018年度学术新苗培养及创新探索专项项目[黔科合平台人才(2018)5779-38]

作者(Author): 范友羊,刘俊,周海燕,曾家顺,李龙
FAN Youyang,LIU Jun,ZHOU Haiyan,ZENG Jiashun,LI Long

DOI: 10.19367/j.cnki.2096-8388.2020.09.002

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