摘要
背景:类风湿关节炎是以小关节疼痛肿胀及晨僵为主要临床表现的自身免疫性疾病,目前药物治疗主要通过非类固醇抗炎药联合使用改善病情抗风湿药物,但其药物对T细胞亚群平衡的影响及其机制目前尚不清楚。目的:探讨甲氨蝶呤联合双氯芬酸钠治疗胶原诱导性关节炎(CIA)大鼠对Th17/Treg细胞平衡的影响及其作用机制。方法:建立雄性Wistar大鼠胶原诱导性关节炎模型,随机分为空白组、模型组、甲氨蝶呤组、双氯芬酸钠组、甲氨蝶呤联合双氯芬酸钠组(联合用药组)、阳性对照组。通过每周评定关节炎指数、测量足趾肿胀程度判断造模成功。药物治疗4周后取材,酶联免疫吸附实验检测外周血中白细胞介素10、转化生长因子β等细胞因子表达情况;流式细胞术检测脾脏Th17及Treg细胞的比例变化情况;RT-PCR法分别检测脾脏中Th17、Treg细胞的转录因子ROR-γt及Foxp3的mRNA表达。结果与结论:(1)药物治疗后,甲氨蝶呤组、双氯芬酸钠组、联合用药组与模型组比较,各治疗组均可改善大鼠一般状况及足趾肿胀程度、降低关节炎指数评分、减少炎症细胞浸润、抑制转录因子ROR-γt调控Th17细胞分化、调控转录因子Foxp3诱导CD4^+CD25+Foxp3^+Treg高分化,从而恢复Treg/Th17比例平衡,改善胶原诱导性关节炎大鼠关节炎症状,各组间平均灰度值差异有显著性意义(P <0.05);(2)联合组关节炎指数下降趋势更明显,且关节炎症状轻于其他造模组;双氯芬酸钠组能上调模型组大鼠外周血白细胞介素10、白细胞介素6表达,甲氨蝶呤组下调转化生长因子β水平占优势,联合用药组能显著上调模型组血清白细胞介素6、白细胞介素10水平,下调转化生长因子β表达;联合用药组能有效上调模型组Treg/Th17比例,显著抑制转录因子Foxp3及ROR-γt表达;(3)结果提示甲氨蝶呤联合双氯芬酸钠治疗具有协同效应,为治疗类风湿关节�
BACKGROUND:Rheumatoid arthritis is an autoimmune disorder that typically results in swollen and painful facet joints and morning stiffness.Its drug therapy is mainly the combination of non-steroidal anti-inflammatory drugs and anti-rheumatic drugs,however,the drug effects on the T-lymphocyte subset balance and the underlying mechanism remain unclear.OBJECTIVE:To explore the effect of the combination of methotrexate and diclofenac sodium on the balance of Th17/Treg cells in a rat model of collagen-induced arthritis.METHODS:After the male Wistar rat model of collagen-induced arthritis model was established,they were randomly divided into blank control,model,methotrexate,diclofenac sodium,methotrexate+diclofenac sodium,and positive control groups.They were assessed by measuring the arthritis index and toe swelling degree.After 4-week treatment,the expression levels of interleukin 10 and transforming growth factorβin the peripheral blood was detected by ELISA,and the change in Th17/Treg cells in the spleen was tested by flow cytometry;the mRNA expression levels of Foxp3 and ROR-γt in the spleen were examined by RT-PCR.RESULTS AND CONCLUSION:Compared with the model group,methotrexate,diclofenac sodium,and their combination could improve the general condition,reduce toes swelling degree,lower arthritis index(especially the methotrexate+diclofenac sodium group),reduce inflammation cell infiltration,and inhibit ROR-γt regulating Th17 cell differentiation and regulating the Foxp3-induced high differentiation of CD4^+CD25^+Foxp3^+Tre,thereby restoring Treg/Th17 cell balance and improving the arthritis symptoms.The average gray value was significantly different among groups(P<0.05).Except the down-regulation of interleukin 10 and interleukin 6 levels like diclofenac sodium,the combined therapy could down-regulate the expression level of transforming growth factorβ,increase the Treg/Th17 cell proportion,and inhibit the expression of Foxp3 and ROR-γt.Additionally,methotrexate significantly reduced the expression level of
作者
牛梓晗
冉新建
麦伍拉尼·马木提
潘传鹏
何东航
任聪改
徐琦
Niu Zi-han;Ran Xin-jian;Mawlan·Mamut;Pan Chuan-peng;He Dong-hang;Ren Cong-gai;Xu Qi(School of Clinical Medicine,Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Laboratory Center of Mechanism,School of Basic Medicine,Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;School of Basic Medicine,School of Basic Medicine,Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Department of Immunolgy,School of Basic Medicine,Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第32期5163-5168,共6页
Chinese Journal of Tissue Engineering Research
基金
国家级大学生创新性实验计划重点项目(CX 201610760022)。
