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长链非编码TRPM2-AS通过靶向miR-545-3p/Bmp2调控骨折愈合。

Long non-coding TRPM2-AS regulates fracture healing by targeting miR-545-3p/Bmp2.

作者信息

Kang Renjie, Huang Lina, Zeng Teng, Ma Jinliang, Jin Danjie

机构信息

Department of Orthopedics, Peking University First Hospital Taiyuan Hospital, Taiyuan, 030000, China.

Department of Rehabilitation Medicine, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, 533000, China.

出版信息

J Orthop Surg Res. 2024 Aug 8;19(1):466. doi: 10.1186/s13018-024-04969-5.

DOI:10.1186/s13018-024-04969-5
PMID:39118176
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11308420/
Abstract

OBJECTIVE

Delayed fracture healing increases the suffering of patients. An in-depth investigation of the pathogenesis of delayed fracture healing may offer new direction for the prevention and treatment.

METHODS

The study included 63 normal healing tibial fractures and 58 delayed healing tibial fractures patients. Long non-coding RNA (lncRNA)TRPM2-AS, microRNA-545-3p (miR-545-3p), bone morphogenetic protein 2 (Bmp2) mRNA and osteogenic differentiation markers, including runt-related transcription factor 2 (Runx2), osteocalcin (Ocn), and alkaline phosphatase (Alp) mRNA expression were determined by Real-time quantitative reverse transcription-polymerase chain reaction in serum and MC3T3-E1 cells. The prediction potential of TRPM2-AS in delayed healing fracture patients was verified by receiver operating characteristic curves. The binding relationship of TRPM2-AS/miR-545-3p/Bmp2 was evaluated by dual luciferase reporter gene assay. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry.

RESULTS

TRPM2-AS was remarkably down-regulated in patients with delayed fracture healing and could better predict the fracture healing status. TRPM2-AS downregulation inhibited osteogenic markers mRNA expression, restrained proliferation, and promoted apoptosis of MC3T3-E1 cells (p < 0.05). In delayed fracture healing, miR-545-3p was dramatically up-regulated and was negatively regulated by TRPM2-AS. Reducing miR-545-3p eliminate the negative effect of TRPM2-AS down-regulation on osteoblast proliferation and differentiation (p < 0.05). miR-545-3p targets Bmp2, which plays a positive role in osteoblast differentiation (p < 0.05).

CONCLUSION

This study found that TRPM2-AS has the potential to be a diagnostic marker for delayed fracture healing and revealed that the TRPM2-AS/miR-545-3p/Bmp2 axis affects fracture healing by regulating osteoblast.

摘要

目的

骨折愈合延迟增加了患者的痛苦。深入研究骨折愈合延迟的发病机制可能为预防和治疗提供新方向。

方法

本研究纳入63例正常愈合的胫骨骨折患者和58例骨折愈合延迟的胫骨骨折患者。采用实时定量逆转录-聚合酶链反应检测血清和MC3T3-E1细胞中长链非编码RNA（lncRNA）TRPM2-AS、微小RNA-545-3p（miR-545-3p）、骨形态发生蛋白2（Bmp2）mRNA以及成骨分化标志物，包括 runt相关转录因子2（Runx2）、骨钙素（Ocn）和碱性磷酸酶（Alp）mRNA的表达。通过受试者工作特征曲线验证TRPM2-AS在骨折愈合延迟患者中的预测潜力。采用双荧光素酶报告基因检测法评估TRPM2-AS/miR-545-3p/Bmp2的结合关系。通过CCK-8法和流式细胞术检测细胞增殖和凋亡情况。

结果

骨折愈合延迟患者中TRPM2-AS显著下调，且能更好地预测骨折愈合状态。TRPM2-AS下调抑制了成骨标志物mRNA表达，抑制了MC3T3-E1细胞的增殖并促进其凋亡（p＜0.05）。在骨折愈合延迟过程中，miR-545-3p显著上调，且受TRPM2-AS负调控。降低miR-545-3p可消除TRPM2-AS下调对成骨细胞增殖和分化的负面影响（p＜0.05）。miR-545-3p靶向Bmp2，Bmp2在成骨细胞分化中起积极作用（p＜0.05）。

结论

本研究发现TRPM2-AS有可能成为骨折愈合延迟的诊断标志物，并揭示了TRPM2-AS/miR-545-3p/Bmp2轴通过调节成骨细胞影响骨折愈合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e768/11308420/81622e8266ad/13018_2024_4969_Fig7_HTML.jpg
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