Fang Shanhong, You Mengqiang, Wei Jie, Chen Peng
Department of Sports Medicine, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
Department of Orthopedic Surgery, the First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
Turk J Biol. 2023 May 18;47(3):186-198. doi: 10.55730/1300-0152.2654. eCollection 2023.
Osteonecrosis of the femoral head (ONFH) is an ischemic disease characterized by the impairment of angiogenesis. We have previously elucidated the role of tsRNAs and BMSC exosomes in ONFH, but whether tsRNA-modified BMSC exosomes promote angiogenesis in ONFH remains unclear.
The expression of angiogenesis-related tsRNA in plasma exosomes from ONFH patients was examined by q-PCR. The function of tsRNA in HUVECs was identified by CCK-8 and angiogenesis assay. Exosomes purified from tsRNA-15797 overexpressed BMSCs were cocultured with HUVECs to examine their role in angiogenesis. The molecule mechanism of tsRNA-15797-modified exosomes was explored by RNA sequencing, dual-luciferase assay, and immunofluorescence.
A tRNA-derived small RNA tsRNA-15797 was down-regulated in plasma exosomes of ONFH patients. We found the effects of BMSCs-derived exosomes on accelerating HUVECs angiogenesis and migration, which were further enhanced after overexpressing tsRNA-15797. Besides, overexpression of tsRNA-15797 would lead to down-regulation of LFNG correlated with angiogenesis. tsRNA-15797 could directly interact with LFNG. We demonstrated that LNFG overexpression weakened the pro angiogenic and migratory effects of tsRNA-15797-modified BMSCs-derived exosomes.
We successfully constructed tsRNA-15797-modified BMSC-derived exosomes and demonstrated that it induced the angiogenesis of HUVECs by targeting the down-regulation of LFNG. Thus, tsRNA-15797-loaded BMSCs-derived exosomes may be a potential target therapy drug for ONFH.
股骨头坏死（ONFH）是一种以血管生成受损为特征的缺血性疾病。我们之前已经阐明了tsRNAs和骨髓间充质干细胞（BMSC）外泌体在ONFH中的作用，但tsRNA修饰的BMSC外泌体是否能促进ONFH中的血管生成仍不清楚。
通过q-PCR检测ONFH患者血浆外泌体中血管生成相关tsRNA的表达。通过CCK-8和血管生成试验鉴定tsRNA在人脐静脉内皮细胞（HUVEC）中的功能。将从过表达tsRNA-15797的BMSCs中纯化的外泌体与HUVECs共培养，以检查它们在血管生成中的作用。通过RNA测序、双荧光素酶测定和免疫荧光探索tsRNA-15797修饰外泌体的分子机制。
一种源自tRNA的小RNA tsRNA-15797在ONFH患者的血浆外泌体中表达下调。我们发现BMSCs来源的外泌体对加速HUVECs血管生成和迁移有作用，在过表达tsRNA-15797后这种作用进一步增强。此外，tsRNA-15797的过表达会导致与血管生成相关的LFNG下调。tsRNA-15797可直接与LFNG相互作用。我们证明LFNG的过表达减弱了tsRNA-15797修饰的BMSCs来源外泌体的促血管生成和迁移作用。
我们成功构建了tsRNA-15797修饰的BMSC来源外泌体，并证明它通过靶向LFNG的下调诱导HUVECs的血管生成。因此，负载tsRNA-15797的BMSCs来源外泌体可能是ONFH的一种潜在靶向治疗药物。
