[关键词]
[摘要]
目的 探讨循环血miR-345-3p在长骨感染性骨不连中的表达变化,评估其作为潜在的新型生物标志物在长骨感染性骨不连的诊断价值。方法 建立股骨感染性骨不连及开放性截骨大鼠模型,术后42 d后,拍摄X线评估内固定位置及骨折愈合情况;离断股骨后行micro-CT,检测骨痂矿物质密度(BMD)、骨痂体积/总体积(BV/TV)、骨小梁厚度(Tb.Th)、骨小梁数量(Tb.N)、骨小梁间隙(Tb.Sp)。使用Sysmex XN-350全血细胞分析仪进行白细胞计数(WBC);采用免疫散射比浊法测定血中C反应蛋白(CRP)表达水平;采用毛细管动态光度计成像测定法检测红细胞沉降速率(ESR)水平。通过实时荧光定量PCR(qRT-PCR)检测和比较感染性骨不连组及骨折组全血中miR-345-3p的表达水平,并通过受试者工作特征(ROC)曲线评估WBC、CRP、ESR及miR-345-3p在感染性骨不连中的诊断效能。结果 行X线及micro-CT评估骨折愈合情况,骨折组骨折线模糊,断端连续,有连续骨痂通过骨折线;长骨感染性骨不连组骨折端硬化,断端有间隙,骨痂间无小梁通过,骨膜增厚。感染性骨不连组术后42 d,BMD、Tb.Th、Tb.N显著高于骨折组,Tb.Sp显著低于骨折组(P<0.05、0.01)。感染性骨不连组WBC、CRP、ESR较骨折组显著升高(P<0.05),miR-345-3p表达较骨折组显著降低(P<0.01)。miR-345-3p诊断的AUC为0.906,优于WBC、CRP、ESR。结论 全血miR-345-3p在长骨感染性骨不连中表达水平显著降低,有望成为感染性骨不连诊断的潜在新型生物标志物。
[Key word]
[Abstract]
Objective To investigate the expression changes of miR-345-3p in circulating blood and evaluate its diagnostic value as a potential new biomarker in infected nonunion of the long bones. Methods The rat models of femoral infected nonunion and open fracture were established. After 42 days of operation, X-ray was taken to evaluate the position of internal fixation and fracture healing; micro CT was performed to detect the bone mineral density (BMD), bone volume/total volume (BV/TV), trabecular thickness (Tb. Th), trabecular number (Tb. N) and trabecular separation (Tb. Sp) of callus. White blood cell count (WBC) was measured by Sysmex XN-350 whole blood cell analyzer, C-reactive protein (CRP) expression was measured by immunonephelometry, erythrocyte sedimentation rate (ESR) was measured by capillary dynamic photometer. Quantitative real time reverse transcription polymerase chain reaction (qRT-PCR) was used to detect and compare the expression level of miR-345-3p in the whole blood of the infected nonunion group and the fracture group. The diagnostic efficacy of WBC, CRP, ESR, and miR-345-3p in infected nonunion were evaluated by the receiver operating characteristic (ROC) curve. Results X-ray and micro CT were used to evaluate the fracture healing. In the fracture group, the fracture line was fuzzy, the fracture end was continuous, and there was a continuous callus passing through the fracture line. In the long bone infectious nonunion group, the fracture end was sclerotic, the fracture end had a gap, there was no trabecula passing through between the callus, and the periosteum was thickened. 42 days after operation, BMD, Tb.Th and Tb.N were significantly higher and Tb.Sp was significantly lower in infected nonunion group than in fracture group (P<0.05, 0.01). WBC, CRP and ESR were significantly higher and miR-345-3p expression was significantly lower in infected nonunion group than in fracture group (P<0.05, 0.01). The AUC of miR-345-3p was 0.906, which was better than WBC, CRP and ESR. Conclusion The expression level of miR-345-3p in infected nonunion of the long bones was reduced, which may be a potential new biomarker for the diagnosis of infected nonunion.
[中图分类号]
R965
[基金项目]
国家自然科学基金资助项目(81603092);江苏省卫计委"科教强卫工程"青年医学人才项目(QNRC2016606)