罗鹏 郑远 葛海峰

[摘要] 目的 采用液相色譜-质谱联用技术的代谢组学方法，探讨非创伤性股骨头坏死（NONFH）患者血浆来源微泡的生物标志物，为NONFH的诊断提供依据。 方法 选取2020年5—11月温州医科大学附属第二医院收治的20例NONFH患者及配对的20名健康体检者的血浆样本，分离微泡，并采用超高效液相色谱串联飞行时间质谱法分析微泡中代谢物，结合主成分分析法（PCA）和正交偏最小二乘分辨分析法（OPLS-DA）筛选差异性代谢产物。结果 样本中共鉴定出12种差异性代谢产物，其中肌苷、次黄嘌呤、尿酸及花生四烯酸上调（P<0.05）;半胱氨酸、同型半胱氨酸、丙酮酸、苹果酸、α-酮戊二酸、硫酸脱氢表雄酮（DHEA-S）和硫酸孕烯醇酮下调（P<0.05）。 结论 采用超高效液相色谱串联飞行时间质谱法分析NONFH血浆来源微泡的代谢产物是可行的，为NONFH的筛查提供新的参考依据。

[关键词] 股骨头坏死;微泡;代谢组学;创伤

[中图分类号] R681.8          [文献标识码] A          [文章编号] 1673-9701（2021）35-0032-05

Metabonomics study of plasma-derived microvesicles in patients with non-traumatic necrosis of the femoral head

LUO Peng1   ZHENG Yuan2   GE Haifeng3

1.Department of Orthopedics，the Second Affiliated Hospital and  Children′s Hospital of Wenzhou Medical University， Wenzhou   325000，China; 2.The Second School of Medicine of Wenzhou Medical University， Wenzhou   325000， China; 3.Department of Laboratory Medicine，the Second Affiliated Hospital and Children′s Hospital of Wenzhou Medical University，Wenzhou   325000，China

[Abstract] Objective To explore the biomarkers of plasma-derived microvesicles from patients with non-traumatic necrosis of the femoral head （NONFH） by using the metabonomics method of liquid chromatography-mass spectrometry， and to provide a basis for the diagnosis of NONFH. Methods A total of 20 patients with NONFH and 20 matched healthy subjects who were admitted to the Second Affiliated Hospital of Wenzhou Medical University From May to November 2020 were selected. The microbubbles were separated. The metabolites in microvesicles were analyzed using ultra-high performance liquid chromatography tandem time of flight mass spectrometry. The differential metabolites were screened combined with principal component analysis （PCA） and orthogonal partial least squares resolution analysis （OPLS-DA）. Results A total of 12 different metabolites were identified in the sample， of which inosine，hypoxanthine，uric acid and arachidonic acid were up-regulated （P<0.05）;cysteine，homocysteine，pyruvate，malic acid，α-ketoglutarate， dehydroepiandrosterone sulfate （DHEA-S） and pregnenolone sulfate were down-regulated（P<0.05）. Conclusion It is feasible to analyze the metabolites of NONFH plasma-derived microbubbles by ultra-high performance liquid chromatography tandem time-of-flight mass spectrometry，which provides a new reference basis for NONFH screening.

[Key words] Femoral head necrosis; Microvesicles; Metabolomics; Trauma

非創伤性股骨头坏死（Non traumatic osteonecrosis of the femoral head，NONFH）常与长期、大剂量或不规范使用糖皮质激素、大量酗酒等有关，约占股骨头坏死总数的70%～80%[1]。现有的多种影像学方法虽使得NONFH的早期诊断率得以提升，但各类影像学方法仍存在缺陷，使得该病的早期诊断依旧困难重重。微泡（Microvesicles，MVs）是一组由多种细胞在生理和病理状态下释放的，直径在100～1000 nm之间的异质性膜囊泡[2]。近年来，随着研究的深入，人们逐渐认识到微泡在疾病诊断、预后及个性化治疗方面具有广阔的医学应用前景[3]。本研究采用代谢组学的方法对健康人群及非创伤性股骨头坏死患者血浆来源的微泡进行分析，找寻差异性代谢产物，筛选出潜在的生物标记物，为疾病的诊断提供新的依据，现报道如下。……