基于Wnt3a/β-catenin通路探究下调miR-16对类风湿性关节炎模型大鼠的干预效果  

The intervention effect of down regulated miR-16 on rheumatoid arthritis model rats based on Wnt3a/β-catenin pathway

作　　者：崔树北[1] 颜继英[1] 贾晓川[1] 仝路[1] 李毅 张开 刘振武[1] Cui Shubei;Yan Jiying;Jia Xiaochuan;Tong Lu;Li Yi;Zhang Kai;Liu Zhenwu(First Department of Orthopedics,Handan Central Hospital,Handan 056000)

机构地区：[1]邯郸市中心医院骨一科,邯郸056000

出　　处：《解剖学杂志》2023年第1期34-38,共5页Chinese Journal of Anatomy

基　　金：河北省医学科学研究重点课题(20181674)。

摘　　要：目的 :探究下调miR-16对类风湿性关节炎模型大鼠的干预效果及其作用机制。方法 :将SPF级SD大鼠随机分为对照组、模型组、阴性对照组(NC)和实验组。通过全弗氏佐剂(CFA)与Ⅱ型胶原乙酸溶液注射关节构建模型组、NC组与实验组建立大鼠类风湿关节炎模型,造模后NC组和实验组分别经尾静脉注射5 nmol/L NC-antagomiR和miR-16-antagomiR,对照组和模型组给予等量生理盐水,以足趾肿胀度为模型标准。qPCR检测各组大鼠滑膜组织miR-16表达水平,对大鼠膝关节进行H-E染色分析并进行病理评分,ELISA检测大鼠血清白介素1β(IL-1β)、白介素-6(IL-6)、白介素17(IL-17)、肿瘤坏死因子α(TNF-α)水平,免疫印迹检测大鼠滑膜组织Wnt家庭成员3a(Wnt3a)和β-连环蛋白(β-catenin)蛋白表达水平。结果 :qPCR结果显示,与对照组相比,模型组和NC组miR-16显著上调,实验组显著下调。与对照组相比,模型组、NC组、实验组大鼠足趾肿胀度及病理评分均显著升高;与模型组和NC组相比,实验组足趾肿胀度及病理评分显著降低。ELISA结果显示,与对照组相比,模型组、NC组、实验组血清IL-1β、IL-6、IL-17、TNF-α显著升高;与模型组和NC组相比,实验组显著降低,但仍高于对照组。免疫印迹结果显示,与对照组相比,模型组和NC组Wnt3a及β-catenin显著升高,而实验组显著降低。结论 :下调miR-16可抑制类风湿性关节炎模型大鼠炎症因子释放,降低组织炎性损伤,发挥骨保护作用,其机制可能与Wnt3a/β-catenin信号通路的调控相关。Objective:To investigate the intervention effect and mechanism of down regulated miR-16 on rheumatoid arthritis model rats.Methods:SD rats were randomly divided into a control group,a model group,an NC group,and an experimental group.Rats with rheumatoid arthritis were established through injecting CFA and collagenⅡvia the caudal vein.In the NC group and experimental group,5 nmol/L NC antagomiR and miR-16-antagomiR were injected respectively,and the control group and model group were given the same amount saline.The toe swelling degree was taken as the model standard.qPCR was used to detect the expression level of miR-16 in synovial tissue of rats in each group.H-E staining was used to analyze the knee joint of rats and pathological score.ELISA was used to detect interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-17(IL-17)and tumor necrosis factorα(TNF-α)in peripheral blood.Expression of Wnt family member 3A(Wnt3a)andβ-catenin protein were detected by Western blotting.Results:qPCR showed that compared with the control group,miR-16 up-regulated significantly in the model group and NC group,but down-regulated in the experimental group.Compared with the control group,the toe swelling degree and pathological score of rats in the model group,NC group and experimental group increased significantly while decreased significantly in the experimental group compared to the model and NC groups.ELISA showed that the peripheral blood IL-1β,IL-6,IL-17,TNF-αincreased in the NC group,model group and experimental group compared to the control group while the experimental group decreased significantly compared with the model group and NC group.Western blotting showed that compared with the control group,Wnt3a andβ-catenin increased significantly in the model group and NC group but decreased significantly in the experimental group.Conclusion:Down regulation of miR-16 can inhibit the release of inflammatory factors,reduce pathological injury,and play a protective role on bones of rheumatoid arthritis model rats.The mecha

关 键 词：miR-16 类风湿性关节炎 炎症 WNT3A Β-连环蛋白 大鼠 

分 类 号：R593.22[医药卫生—内科学] R-332[医药卫生—临床医学]