长链非编码RNA胰岛素样生长因子2-AS对大鼠骨质疏松性骨折的影响及其机制  

Effects of long non-coding RNA insulin-like growth factor 2-AS on osteoporotic fracture in rats and its mechanism

作　　者：付永良[1] 郝耀 赵巍[1] 李辉 赵锦浩 Fu Yongliang;Hao Yao;Zhao Wei;Li Hui;Zhao Jinhao(Department of Orthopedics,First Hospital of Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学第一医院骨科,太原030001

出　　处：《中华实验外科杂志》2024年第5期1029-1032,共4页Chinese Journal of Experimental Surgery

基　　金：山西省应用基础研究计划(201901D211481)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)胰岛素样生长因子2(IGF2)-AS对骨质疏松性大鼠骨折愈合的影响及其分子机制。方法:30只SD大鼠按照随机数字表法分为对照组、骨折组和lncRNA IGF2-AS组,骨折组和lncRNA IGF2-AS组大鼠采用去卵巢手术建立骨质疏松大鼠模型,并在去卵巢后3个月后建立胫骨骨折模型,对照组大鼠仅去卵巢,而胫骨未骨折。lncRNA IGF2-AS组大鼠在去卵巢后2个月在胫骨注射过表达lncRNA IGF2-AS的腺相关病毒,对照组和骨折组注射空载体腺相关病毒。在建模4周后,观察3组大鼠骨折愈合情况和骨密度变化;采用蛋白质免疫印迹(Western blot)分析骨痂组织骨形成蛋白-2、成骨细胞特异蛋白(Osterix)和成骨细胞分化因子(ODF)表达水平;生物力学分析3组大鼠胫骨生物力学变化;采用骨组织形态计量学分析矿化宽度、矿化体积、类骨质宽度和体积变化。组间计量数据采用单因素方差分析。结果:对照组大鼠胫骨完整,无骨折;骨折组大鼠骨折线明显,骨折端少量稀疏骨痂;lncRNA IGF2-AS组大鼠骨折端外骨痂致密,骨折线模糊。lncRNA IGF2-AS组大鼠胫骨骨密度[(0.087±0.003)g/cm 2]明显高于骨折组大鼠[(0.072±0.008)g/cm 2],差异有统计学意义(t=5.486,P<0.05)。lncRNA IGF2-AS组大鼠胫骨骨密度[(0.087±0.003)g/cm 2]明显高于骨折组大鼠[(0.072±0.008)g/cm 2],差异有统计学意义(t=5.486,P<0.05)。骨形成蛋白-2、Osterix和ODF表达水平(1.50±0.14、1.47±0.20、1.61±0.11)明显高于骨折组大鼠(0.64±0.11、0.53±0.09、0.73±0.10),差异有统计学意义(t=15.310、13.610、19.480,P<0.05)。lncRNA IGF2-AS组大鼠最大载荷、剪切强度和弹性模量[(32.53±2.65)N、(60.37±2.97)Kn/mm 2、(182.89±15.06)GPa]明显高于骨折组大鼠[(27.32±3.35)N、(48.06±4.97)Kn/mm 2、(148.97±14.75)GPa],差异有统计学意义(t=3.886、6.727、5.088,P<0.05)。lncRNA IGF2-AS组大鼠骨小梁相对体积、骨小梁平均骨壁厚度和矿化延Objective To investigate the effects of long non-coding RNA(lncRNA)insulin-like growth factor 2(IGF2)-AS on fracture healing in osteoporotic rats and its molecular mechanism.Methods Totally,30 SD rats were divided into control group,fracture group and lncRNA IGF2-AS group according to the random number table method.Rat models of osteoporosis were established by ovariectomy in the fracture group and lncRNA IGF2-AS group,and tibial fracture models were established at 3rd month after ovariectomy in the control group.Rats in the lncRNA IGF2-AS group were injected with adeno-associated virus overexpressing lncRNA IGF2-AS in the tibia at 2nd month after ovariectomy,and those in the control group and fracture group were injected with empty adeno-associated virus.After 4 weeks of modeling,the fracture healing and bone mineral density changes of the three groups were observed.The expression levels of bone morphogenetic protein-2,osteoblast-specific protein(Osterix)and osteoblast differentiation factor(ODF)in the callus tissue were analyzed by Western blotting.The biomechanical changes of the tibia of the three groups were analyzed by biomechanics.The changes of mineralized width,mineralized volume,osteoid width and volume were analyzed by bone tissue morphometry.The measurement data between groups were analyzed by one-way analysis of variance.Results The tibia of rats in the control group was intact without fracture.The fracture line of rats in the fracture group was obvious,and there was a small amount of sparse callus at the fracture end.The callus outside the fracture end of rats in the lncRNA IGF2-AS group was dense,and the fracture line was blurred.The tibial bone mineral density of rats in lncRNA IGF2-AS group[(0.087±0.003)g/cm2]was significantly higher than that in the fracture group[(0.072±0.008)g/cm2,t=5.486,P<0.05].The expression levels of bone morphogenetic protein-2,osteoblast-specific protein(Osterix)and osteoblast differentiation factor(ODF)in rats in lncRNA IGF2-AS group(1.50±0.14,1.47±0.20,1.61±0.11)w

关 键 词：长链非编码RNA 胰岛素样生长因子 骨质疏松性骨折 骨折愈合 生物力学 

分 类 号：R683[医药卫生—骨科学]