GLP-1/GLP-1R通路介导的内质网应激在大鼠骨质疏松性骨折愈合中的作用机制  

Mechanism of Endoplasmic Reticulum Stress Mediated by GLP-1/GLP-1R Pathway in the Healing of Osteoporotic Fractures in Rats

作　　者：陈艳 陈卫红 席薇 杨东海 银晓永 CHEN Yan;CHEN Wei-hong;XI Wei;YANG Dong-hai;YIN Xiao-yong(Department of Integrated Traditional Chinese and Western Medicine Orthopedics,the First Hospital of Handan City;Orthopedics Department,North China Medical Fengfeng General Hospital at Handan,Handan 056000,China)

机构地区：[1]邯郸市第一医院中西医结合骨科 [2]华北医疗峰峰总医院邯郸院区骨科,河北邯郸056000

出　　处：《南昌大学学报(医学版)》2025年第1期6-11,共6页Journal of Nanchang University:Medical Sciences

基　　金：河北省卫生健康委员会科研项目(20220012)。

摘　　要：目的探讨胰高糖素样肽-1(glucagon-like peptide-1,GLP-1)/GLP-1受体(GLP-1 receptor,GLP-1R)通路介导的内质网应激在大鼠骨质疏松性骨折(OF)愈合中的可能作用机制。方法将30只大鼠随机分为3组:假手术组、OF组、GLP-1R激活组,每组10只,OF组、GLP-1R激活组建立大鼠OF模型,GLP-1R激活组大鼠建模成功后给予GLP-1R激活剂灌胃处理。采用免疫组织化学染色检测骨组织中GLP-1R表达水平,ELISA试剂盒检测血清GLP-1及骨代谢相关指标[碱性磷酸酶(ALP)、骨钙素(BGP)、Ⅰ型胶原氨基端前肽(PINP)、N-端交联肽(NTx)及酸性磷酸酶(TRAP)]的含量,双能X线骨密度仪检测大鼠股骨骨密度。采用Western blotting及RT-qPCR检测内质网应激相关蛋白(GRP78、Caspase-12、CHOP、PERK蛋白及mRNA)水平。采用Micro-CT三维重建股骨断端愈合情况。采用HE染色检测骨及软骨组织病理变化。结果与假手术组相比,OF组大鼠血清GLP-1、ALP、BGP、PINP水平显著减少,NTx、TRAP水平显著增加(P<0.05);骨折端骨组织GLP-1R表达水平显著减少,GRP78、Caspase-12、CHOP、PERK蛋白及mRNA水平显著增加(P<0.05);股骨骨密度显著减少(P<0.05)。与OF组比较,GLP-1R激活组大鼠血清ALP、BGP、PINP水平显著增加,NTx、TRAP水平显著减少(P<0.05);骨折端骨组织GRP78、Caspase-12、CHOP、PERK蛋白及mRNA水平显著减少(P<0.05);股骨骨密度显著增加(P<0.05)。OF组与GLP-1R激活组大鼠骨折断端缝隙均逐渐缩小、愈合,GLP-1R激活组比OF组大鼠股骨断端随时间增加愈合速度增加,愈合效果更好。结论激活GLP-1/GLP-1R通路可抑制OF大鼠断端骨组织中内质网应激水平,促进骨质疏松性骨折愈合。Objective To explore the potential mechanism of endoplasmic reticulum stress mediated by the glucagon-like peptide-1(GLP-1)/GLP-1 receptor(GLP-1R)in the healing of osteoporotic fractures(OF)in rats.Methods 30 rats were randomly divided into 3 groups:sham operation group,OF group,and GLP-1R-activated group,with 10 rats in each group.OF models were established in both OF group and GLP-1R-activated group,and rats in the GLP-1R-activated group were administered with GLP-1R activator by gavage after successful modeling.The expression level of GLP-1R in bone tissue was detected by immunohistochemical staining;ELISA kits were employed to measure the concentrations of serum GLP-1 and bone metabolism-related indicators,including total alkaline phosphatase(ALP),bone gla protein(BGP),procollagen type I N-terminal propeptide(PINP),N-terminal telopeptide(NTx),and tartrate-resistant acid phosphatase(TRAP);dual-energy X-ray bone densitometer was utilized to assess the bone mineral density of the rat femur.Western blotting and RT-qPCR were utilized to measure the levels of endoplasmic reticulum stress-related proteins(GRP78,Caspase-12,CHOP,PERK proteins and mRNA);Micro-CT was employed to reconstruct the healing process of femur fractures in three dimensions;HE staining was conducted to examine pathological changes in bone and cartilage tissue.Results Compared to the sham operation group,the OF group rats exhibited significantly reduced levels of serum GLP-1,ALP,BGP,and PINP,while the levels of NTx and TRAP were significantly increased(P<0.05);there was a notable decrease in GLP-1R expression in the bone tissue at the fracture site,accompanied by a significant increase in GRP78,Caspase-12,CHOP,and PERK protein and mRNA levels(P<0.05);a significant reduction in femoral bone mineral density was observed(P<0.05).Compared to the OF group,the GLP-1R-activated group showed significantly increased levels of serum ALP,BGP,and PINP,while the levels of NTx and TRAP were significantly reduced(P<0.05);there was a notable decrease in the expr

关 键 词：骨质疏松性骨折 GLP-1/GLP-1R通路 内质网应激 骨折愈合 动物 实验 大鼠 

分 类 号：R683.4[医药卫生—骨科学]