甲状旁腺激素1-34通过Wnt/β-catenin信号通路阻断晚期糖基化终末产物对骨髓间充质干细胞负性成骨作用  

PTH1-34 blocks advanced glycation end products′ negative osteogenesis to bone marrow mesenchymal stem cells through Wnt/β-catenin signaling pathway

作　　者：高飞[1] 武珈宇 高虹 董旋 杨静[1] 霍建忠 Gao Fei;Wu Jiayu;Gao Hong;Dong Xuan;Yang Jing;Huo Jianzhong(Department of Endocrinology,the First Hospital of Shanxi Medical University,Taiyuan 030001,China;Intensive Care Unit,the Second Hospital of Shanxi Medical University,Taiyuan 030002,China;Department of Laboratory Diagnostics,Hebei Medical University,Shijiazhuang 050011,China;Department of Orthopaedics,Bethune Hospital of Shanxi,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]山西医科大学第二医院重症医学科,太原030002 [3]河北医科大学实验诊断学教研室,石家庄050011 [4]山西白求恩医院骨科,太原030032

出　　处：《中华内分泌代谢杂志》2020年第6期506-511,共6页Chinese Journal of Endocrinology and Metabolism

基　　金：山西省应用基础研究项目(201901D111351、201901D111409);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：目的观察甲状旁腺激素(PTH)1-34是否可以阻断晚期糖基化终末产物(AGEs)对大鼠骨髓间充质干细胞(BMSCs)的负性成骨作用及其可能信号通路。方法采用全骨髓法分离培养4周龄SD大鼠的BMSCs,分别给予成骨诱导液、牛血清血蛋白(BSA)、AGEs、AGEs联合PTH1-34预处理,7 d后行实时荧光定量PCR(RT-PCR)检测碱性磷酸酶(ALP)、Ⅰ型胶原酶(COL-Ⅰ)mRNA表达水平,Western印迹法检测β-catenin、osterix(OSX)、核心结合蛋白因子2(RUNX2)、终末期氨基糖基化受体蛋白表达水平。21 d后采用茜素红染色检测矿化结节并定量;在10-8 mmol/L的PTH1-34中加入Wnt通路特异性阻断剂Dickkopf-1(DKK1)1μg/ml后,Western印迹法再次检测β-catenin、OSX、RUNX2蛋白表达水平。结果AGEs抑制ALP、COL-ⅠmRNA及β-catenin、OSX、RUNX2蛋白表达(P<0.05),而PTH1-34预处理后逆转AGEs的作用,ALP、COL-ⅠmRNA及β-catenin、OSX、RUNX2蛋白表达较AGEs组增加(P<0.05),茜素红染色矿化结节呈红棕色且增多,定量检测OD值,PTH1-34预处理组高于AGEs组(P<0.05)。DKK1(1μg/ml)作用后,β-catenin、OSX蛋白表达均较BSA组减少(P<0.05)。结论PTH1-34可通过Wnt/β-catenin信号通路阻断AGEs对BMSCs的负性成骨作用。Objective To observe whether parathyroid hormone(PTH)1-34 can block the negative osteogenesis of advanced glycation end products(AGEs)on bone marrow mesenchymal stem cells(BMSCs)in rats and its possible signaling pathways.Methods BMSCs from 4-week-old SD rats were isolated and cultured with whole bone marrow method.The osteogenic induction fluid,bovine serum albumin(BSA),AGEs,AGEs combined with PTH1-34 were pretreated respectively.After 7 days,realtime fluorescence quantitative PCR(RT-PCR)was used to measure alkaline phosphatase(ALP),collagen-Ⅰ(COL-Ⅰ)mRNA expression level,and the expressions ofβ-catenin,osterix(OSX),runt-related transcription factor 2(RUNX2),and receptor for advanced glycation end products protein were examined by Western blotting.Alizarin red staining mineralized nodules and quantitative detection were performed on 21 days.After the addition of Wnt pathway specific blocker Dickkopf-1(DKK1)(1μg/ml)to 10-8 mmol/L of PTH1-34,the protein expression levels ofβ-catenin,OSX,and RUNX2 were detected again by Western blotting.Results AGEs significantly inhibited the expression of ALP,COL-ⅠmRNA,β-catenin,OSX,and RUNX2 proteins(P<0.05).PTH1-34 inhibited AGEs after pretreatment,the expression of ALP,COL-ⅠmRNA,andβ-catenin,OSX,RUNX2 protein were higher than those of AGEs group(P<0.05).The mineralized nodules stained with alizarin red showed reddish brown and increased OD value was detected quantitatively in PTH1-34 group,which was higher than that of AGEs group(P<0.05).DKK1(1μg/ml)reduced the expression ofβ-catenin,OSX proteins as compared with the BSA group(P<0.05).Conclusion PTH1-34 blocks AGEs′negative osteogenesis to BMSCs through Wnt/β-catenin signaling pathway.

关 键 词：甲状旁腺激素1-34 晚期糖基化终末产物 骨髓间充质干细胞 WNT/Β-CATENIN 成骨分化 

分 类 号：R587.2[医药卫生—内分泌]