虎杖苷通过Wnt-Smad通路对强直性脊柱炎小鼠成骨分化的作用机制探究  

Mechanism of polydatin on osteogenesis and differentiation in ankylosing spondylitis mice through Wnt-Smad pathway

作　　者：段婉娥 李俊杰 DUAN Wan'e;LI Junjie(Department of Acupuncture and Rehabilitation,Wuhan Integrated Traditional Chinese and Western Medicine Orthopedic Hospital,and Affiliated Hospital of Wuhan Institute of Physical Education,Wuhan 430070,China;Department of Minimally Invasive Spine Surgery,Wuhan Integrated Traditional Chinese and Western Medicine Orthopedic Hospital,and Affiliated Hospital of Wuhan Institute of Physical Education,Wuhan 430070,China)

机构地区：[1]武汉中西医结合骨科医院暨武汉体育学院附属医院针灸康复科,武汉430070 [2]武汉中西医结合骨科医院暨武汉体育学院附属医院脊柱微创科,武汉430070

出　　处：《中国免疫学杂志》2024年第12期2490-2495,共6页Chinese Journal of Immunology

基　　金：武汉市医学科研项目(WZ20Q05)。

摘　　要：目的:研究虎杖苷(PD)对强直性脊柱炎(AS)小鼠成骨分化的作用以及对Wnt-Smad通路的调控作用。方法:将36只SPF级C57BL/6雄性小鼠随机分为6组:对照组、模型组、阳性组(柳氮磺吡啶组)、PD低、中、高剂量组,每组6只。HE染色检测小鼠骶髂关节滑膜组织病理形态;ELISA检测小鼠血清中TNF-α、MMP-3、OPG含量。分离培养成骨细胞,分组处理,CCK-8检测成骨细胞的增殖活性;试剂盒测定ALP活性;茜素红染色检测矿化能力;Western blot检测Wnt1、β-Catenin、DKK-1、BMP-2、p-Smad1、Runx2蛋白的表达。结果:与对照组相比,模型组小鼠滑膜组织中的毛细血管增多,组织表面肿胀、充血,滑膜细胞不规则排列;血清中TNF-α、MMP-3和OPG含量增加;AS小鼠成骨细胞的存活率、ALP活性、矿化能力以及Wnt1、β-Catenin、BMP-2、p-Smad1、Runx2蛋白的表达升高,DKK-1蛋白表达降低(P<0.05);与模型组相比,阳性组和PD低、中、高剂量组小鼠滑膜组织中的毛细血管增殖减少,组织表面肿胀减轻,滑膜细胞半透明且排列整齐;血清中TNF-α、MMP-3和OPG含量降低;AS小鼠成骨细胞的存活率、ALP活性、矿化能力以及Wnt1、β-Catenin、BMP-2、p-Smad1、Runx2蛋白的表达降低,DKK-1蛋白表达升高(P<0.05)。结论:PD可能通过抑制Wnt-Smad通路的激活,抑制AS小鼠的成骨分化。Objective:To investigate the impact of polydatin(PD)on osteogenesis and differentiation in ankylosing spondylitis(AS)mice and its regulation on Wnt-Smad pathway.Methods:Thirty-six SPF C57BL/6 male mice were randomly divided into 6 groups:control group,model group,positive group(sulfasalazine group),PD low,medium and high doses groups,with 6 mice in each group.HE staining was used to detect the pathological morphology of the synovium of the sacroiliac joint in mice;the contents of TNF-α,MMP-3 and OPG in serum of mice were detected by ELISA.Osteoblasts were isolated and cultured,the proliferation activity of osteoblasts was detected by CCK-8;ALP activity was measured with the kit;Alizarin red staining was used to detect the mineralization ability;Western blot was used to detect the expressions of Wnt1,β-Catenin,DKK-1,BMP-2,p-Smad1 and Runx2 proteins.Results:Compared with control group,the number of capillaries in the synovial tissue of the model group was increased,the surface of the tissue was swollen and congested,and the synovial cells were irregularly arranged;the contents of TNF-α,MMP-3 and OPG in serum were increased;the survival rate,ALP activity,mineralization ability and the expression of Wnt1,β-Catenin,BMP-2,p-Smad1,Runx2 proteins of osteoblasts in AS mice were increased,while the expression of DKK-1 protein was decreased(P<0.05);compared with model group,the proliferation of capillaries in the synovial tissue of the positive group and PD low,medium and high dose groups were decreased,the swelling of the tissue surface was decreased,and the synovial cells were translucent and arranged neatly;the contents of TNF-α,MMP-3 and OPG in serum were decreased;the survival rate,ALP activity,mineralization ability and the expressions of Wnt1,β-Catenin,BMP-2,p-Smad1,Runx2 proteins of osteoblasts in AS mice were decreased,while the expression of DKK-1 protein was increased(P<0.05).Conclusion:PD may inhibit the osteogenic differentiation of AS mice by inhibiting the activation of Wnt-Smad pathway.

关 键 词：虎杖苷 Wnt-Smad通路 强直性脊柱炎 成骨分化 成骨细胞 

分 类 号：R285[医药卫生—中药学]