AGEs对大鼠骨髓间充质干细胞成骨分化中β-catenin、ALP的影响及PTH的干预作用  

Effects of AGEs onβ-catenin and ALP in osteogenic differentiation of rat bone marrow mesenchymal stem cells and the intervention of PTH

作　　者：高飞[1] 董旋 徐倩倩 闫文楠 高虹 霍建忠 杨静[1] Gao Fei;Dong Xuan;Xu Qianqian;Yan Wennan;Gao Hong;Huo Jianzhong;Yang Jing(Department of Endocrinology,The First Hospital of Shanxi Medical University,Taiyuan 030001,China;Department of Experimental Diagnostics,Hebei Medical University,Shijiazhuang 050011,China;Department of Orthopaedics,Shanxi Bethune Hospital,Taiyuan 030032,China)

机构地区：[1]山西医科大学第一医院内分泌科,太原030001 [2]河北医科大学实验诊断学教研室,石家庄050011 [3]山西白求恩医院骨科,太原030032

出　　处：《国际内分泌代谢杂志》2021年第3期173-177,共5页International Journal of Endocrinology and Metabolism

基　　金：山西省应用基础研究项目(201901D111351、20191D111409);山西省重点研发计划项目(201803D31134);山西省卫计委科技攻关项目(201601028)。

摘　　要：目的初步探讨在培养的骨髓间充质干细胞(BMSCs)层面,晚期糖基化终末产物(AGEs)对BMSCs成骨分化中β-catenin、碱性磷酸酶(ALP)的影响,以及甲状旁腺激素(PTH)的干预及其机制。方法取第三代BMSCs,检测AGEs对其增殖的影响。将BMSCs分为对照组、AGEs组、Wnt3a组、AGEs+Wnt3a组;对照组、AGEs组、AGEs+晚期糖基化终末产物受体(RAGE)抗体组;对照组、AGEs组、AGEs+PTH组;对照组、PTH组、PTH+Dickkopf相关蛋白1(DKK1)组,应用Western-blot法检测β-catenin、ALP蛋白的表达。结果BMSCs的增殖可以被AGEs抑制。0.2 g/L AGEs干预BMSCs 7 d,AGEs抑制β-catenin、ALP蛋白表达有统计学差异;给予Wnt通路促进剂Wnt3a后,AGEs作用被抑制,β-catenin、ALP蛋白表达较AGEs组增加;加入10 mg/L RAGE中和抗体共培养,与AGEs组相比,β-catenin、ALP蛋白表达增高。加入PTH后可以逆转AGEs的负性成骨作用,β-catenin、ALP蛋白表达增高;加入Wnt通路阻滞剂DKK1共培养,β-catenin、ALP蛋白表达较PTH组增加。结论AGEs-RAGE通过Wnt/β-catenin信号通路抑制大鼠BMSCs ALP蛋白表达;PTH逆转AGEs对β-catenin、ALP的负性作用。Objective To explore the effect of advanced glycation end products(AGEs)onβ-catenin and alkaline phosphatase(ALP)in osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs),and the intervention of parathyroid hormone(PTH)and its mechanism in cultured BMSCs.Methods The third generation of BMSCs were taken to detect the effect of AGEs on their proliferation.BMSCs were divided into control group,AGEs group,Wnt3a group,AGEs+Wnt3a group;control group,AGEs group,AGEs+RAGE antibody group;control group,AGEs group,AGEs+PTH group;control group,PTH group,PTH+DKK1 group.The expression ofβ-catenin and ALP protein were detected by Western-blot.Results The proliferation of BMSCs was inhibited by AGEs.After 0.2 mg/ml AGEs treatment of BMSCs for 7 days,there was significant difference in the inhibition ofβ-catenin and ALP protein expression by AGEs;after administration of Wnt pathway promoter Wnt3a,the effect of AGEs was inhibited and the expression ofβ-catenin and ALP protein were increased compared with AGEs group;when co-cultured with 10μg/ml RAGE neutralization antibody,the expression ofβ-catenin and ALP protein were increased compared with AGEs group.The negative osteogenic effect of AGEs was reversed and the expression ofβ-catenin and ALP protein were increased by adding PTH,and the expression ofβ-catenin and ALP protein in cells co-cultured with DKK1,a blocker of Wnt pathway,were higher than those in PTH group.Conclusion AGEs-RAGE inhibits the expression of ALP in rat BMSCs through Wnt/β-catenin signal pathway,and PTH reverses the negative effects of AGEs onβ-catenin and ALP.

关 键 词：晚期糖基化终末产物 骨髓间充质干细胞 Β-CATENIN 碱性磷酸酶 甲状旁腺激素 

分 类 号：R587.2[医药卫生—内分泌]