褪黑素对胶原诱导性关节炎大鼠踝关节炎症的作用  

Effect of melatonin on ankle inflammation in collagen-induced arthritis rats

作　　者：田娟 王妍华[1] 胡楠 何岚[1] TIAN Juan;WANG Yan-hua;HU Nan;HE Lan(Department of Rheumatology and Immunology,the First Affiliated Hospital of Xi'anJiaotong University,Xi'an,Shaanri 710061,China)

机构地区：[1]西安交通大学第一附属医院风湿免疫科,陕西西安710061

出　　处：《中华实用诊断与治疗杂志》2022年第8期800-804,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：陕西省自然科学基础研究计划项目(2021JQ-393)。

摘　　要：目的探讨褪黑素对胶原诱导性关节炎大鼠踝关节组织炎症的作用及其对T细胞平衡的影响。方法Wistar大鼠24只,随机分为对照组、模型组、褪黑素组、泼尼松龙组各6只。模型组、褪黑素组、泼尼松龙组大鼠分别于实验第1天、第7天尾部皮下注射200μL牛Ⅱ型胶原蛋白乳剂制备胶原诱导性关节炎大鼠模型,对照组大鼠注射相同剂量生理盐水。褪黑素组给予褪黑素溶液灌胃,1次/d,连续30 d;泼尼松龙组给予泼尼松龙溶液灌胃,1次/d,连续30 d;对照组及模型组大鼠注射相同剂量生理盐水。实验第16天,评估模型组、褪黑素组、泼尼松龙组胶原诱导性关节炎发生情况;实验第30天,4组大鼠取踝关节组织,采用HE染色观察关节组织病理改变情况并对模型组、褪黑素组、泼尼松龙组大鼠踝关节炎症、软骨细胞死亡、软骨表面侵蚀和骨侵蚀行组织病理学评分;应用流式细胞仪检测4组大鼠脾脏组织CD4^(+)CD25^(+)Foxp3^(+)Treg细胞、白细胞介素(interleukin,IL)-4^(+)CD4^(+)T细胞、干扰素(interferon-γ,IFN-γ)^(+)CD4^(+)T细胞表达情况;采用实时荧光定量PCR法检测4组大鼠脾脏组织Foxp3 mRNA相对表达量;采用ELISA法检测4组大鼠踝关节滑液肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、IFN-γ、IL-4、IL-10、IL-17A、IL-6、转化生长因子-β_(1)(transforming growth factor-β_(1),TGF-β_(1))水平。结果实验第16天,模型组胶原诱导性关节炎发生率(83.33%)高于褪黑素组(16.67%)(χ^(2)=5.333,P=0.021),泼尼松龙组(50.00%)与模型组、褪黑素组比较差异均无统计学意义(P>0.05)。实验第30天,模型组踝关节骨质侵蚀、软骨破坏,褪黑素组、泼尼松龙组踝关节骨质均轻度侵蚀,软骨均未见明显异常。模型组踝关节炎症[(2.94±0.16)分]、软骨细胞死亡[(2.78±0.12)分]、软骨表面侵蚀[(2.03±0.10)分]、骨侵蚀[(1.56±0.07)分]组织病理学评分均高于褪黑素组[(1.45±0.06)、(0.54�Objective To investigate the role of melatonin in the inflammation of ankle joint tissue in collagen-induced arthritis rats and its influence on T cell balance.Methods Twenty-four Wistar rats were randomly divided into control group,model group,melatonin group and prednisone group,with 6rats in each group.Rats in model group,melatonin group and prednisolone group were subcutaneously injected with 200μL bovine typeⅡcollagen emulsion on the 1st day and the 7th day of the experiment to prepare the collagen-induced arthritis rat models,and the rats in control group were injected with the same dose of normal saline.Melatonin group was given melatonin solution by gavage,once a day,for 30consecutive days;prednisolone group was given prednisolone solution by gavage,once a day,for 30consecutive days;control group and model group were injected with the same dose of normal saline.On the 16th day of experiment,the occurrence of collagen-induced arthritis in model group,melatonin group and prednisolone group was evaluated.On the30th day of experiment,the ankle joints of four groups were obtained,and HE staining was used to observe the pathological changes of the joint tissues.Histopathological scoring was performed on the ankle joint inflammation,chondrocyte death,cartilage surface erosion and bone erosion in model group,melatonin group and prednisolone group.The expressions of CD4^(+)CD25^(+)Foxp3^(+)Treg,interleukin(IL)-4^(+)CD4^(+),and interferon-γ(IFN-γ)^(+)CD4^(+)T cells in the spleen tissues were detected by flow cytometry.The relative expression of Foxp3 mRNA in the spleen tissues was detected by real-time fluorescence quantitative PCR.ELISA was used to detect the levels of tumor necrosis factor-α(TNF-α),IFN-γ,IL-4,IL-10,IL-17A,IL-6and transforming growth factor(TGF)-β_(1)in the synovial fluid.Results On the 16th day of experiment,the incidence of collagen-induced arthritis was higher in model group(83.33%)than that in melatonin group(16.67%)(χ^(2)=5.333,P=0.021),and showed no significant difference in predni

关 键 词：类风湿关节炎 胶原诱导性关节炎 褪黑素 泼尼松龙 T细胞 炎症 大鼠 

分 类 号：R684.3[医药卫生—骨科学]