壮督驱寒合剂对强直性脊柱炎模型小鼠miR-29a及Wnt信号通路的影响

Effects of Zhuangdu Quhan mixture on miR-29a and Wnt pathway in the ankylosing spondylitis model mice

目的:研究壮督驱寒合剂对强直性脊柱炎(AS)模型小鼠miR-29a及Wnt通路的影响,以探讨壮督驱寒合剂防治AS的机制。方法:将50只6~8周Balb/c雌性小鼠随机分为空白组、模型组、西药组、中药低剂量组、中药高剂量组,每组10只。除空白组外,其余各组均采取蛋白聚糖加完全弗式试剂腹腔注射复制AS小鼠模型。造模结束后各组分别进行相应药物灌胃处理,1次/d,连续6周,末次给药后取膝关节滑膜组织。采用实时荧光定量法测定miR-29a、DKK-1及β-catenin的mRNA表达,采用免疫印迹法检测β-catenin、DKK-1蛋白含量。结果:与空白组相比,模型组小鼠滑膜组织中miR-29a表达升高,β-catenin mRNA及蛋白表达升高,DKK-1 mRNA及蛋白表达下降,差异具有统计学意义(P<0.05)。与模型组相比,西药组及中药低、高剂量组miR-29a表达下降,β-catenin mRNA及蛋白表达下降,DKK-1 mRNA及蛋白表达升高,差异具有统计学意义(P<0.05)。结论:壮督驱寒合剂可能通过调控miR-29a来影响Wnt/β-catenin经典信号通路的表达,从而起到延缓AS新骨形成的作用。

Objective:To study the effect of Zhuangdu Quhan mixture(ZQM)on miR-29a and Wnt pathway in ankylosing spondylitis(AS)model mice,and to explore prevention and treatment mechanism of ZQM for AS.Methods:50 Balb/c mice at 6~8 weeks were randomly divided into normal group,western medicine group,model group,and low,high dose ZQM groups,with 10 mice in each group.Except the normal group,proteoglycan and complete Freund’s reagent were used to establish AS model in each group.After modeling,the corresponding drugs were administered to each group,and the drug was administered once a day,for 6 weeks.The synovial tissue of the knee joint was taken after the last administration.The mRNA expressions of miR-29a,DKK-1 andβ-catenin were determined by real-time fluorescence quantitative method,and the protein contents ofβ-catenin and DKK-1 were detected by western blot.Results:Compared with the normal group,the expression of miR-29a、β-catenin mRNA andβ-catenin protein in the synovial tissue of the model group increased,while the expression of DKK-1 mRNA and DKK-1 protein decreased(P<0.05).Compared with the model group,the expressions of miR-29a,β-catenin mRNA andβ-catenin protein decreased,and the expression of DKK-1 mRNA and DKK-1 protein increased in the western medicine group,and low,high dose ZQM groups(P<0.05).Conclusion:Zhuangdu Quhan mixture may regulate the expression of Wnt/β-catenin signaling pathway through miR-29a,thereby delaying the formation of new bone in AS.