Lai Yu-Cheng, Yao Zhi-Kang, Chang Tien-Chieh, Feng Chien-Wei, Kuo Tsu-Jen, Luo Yi-Wei, Jean Yen-Hsuan, Lin Hugo Y-H, Wen Zhi-Hong
Department of Marine Biotechnology and Resources, National Sun Yat-Sen University, Kaohsiung, Taiwan.
Department of Orthopedics, Asia University Hospital, Taichung, Taiwan.
Environ Toxicol. 2025 Apr;40(4):650-663. doi: 10.1002/tox.24447. Epub 2024 Nov 28.
The glutamatergic signaling pathway, which is mediated by N-methyl-D-aspartate (NMDA) receptors, is crucial for osteoblast differentiation and bone function. Dextromethorphan (DXM), a widely used antitussive, is a noncompetitive antagonist of the NMDA receptor. However, the effects of DXM on osteoblast and bone regeneration remain unclear. The present study investigated the effects of DXM on osteogenesis in vitro and in vivo. A MC3T3-E1 preosteoblast cell line was treated with varying concentrations of DXM. Real-time-quantitative polymerase chain reaction (RT-qPCR) and Western-blot analysis were performed to evaluate the expression of osteogenesis-related runt-related transcription factor 2 (RUNX2), osterix (OSX), osteocalcin (OCN), collagen type 1α (Col-1α), and alkaline phosphatase (ALP) after DXM treatment. Zebrafish embryos were incubated with DXM, which had potential to affect the ossification of the vertebrae and skull, and analyzed using calcein staining. Furthermore, we used a rat calvarial defect model to assess the effects of DXM on bone regeneration by using microcomputed tomography. The results indicate that DXM inhibited extracellular mineralization, ALP activity, and the expression of osteogenic markers, namely RUNX2, OSX, OCN, Col-1α, and ALP, in MC3T3-E1 cells. DXM suppressed skeleton ossification in zebrafish and affected bone regeneration in rats with calvarial defects. However, the mineral density of the regenerated bones did not differ significantly between the DXM and control groups. The present study demonstrated that DXM negatively affects the osteogenic function of osteoblasts, leading to impaired skeletal development and bone regeneration. Thus, clinicians should consider the negative effects of DXM on bone regeneration.
由N-甲基-D-天冬氨酸（NMDA）受体介导的谷氨酸能信号通路对成骨细胞分化和骨功能至关重要。右美沙芬（DXM）是一种广泛使用的止咳药，是NMDA受体的非竞争性拮抗剂。然而，DXM对成骨细胞和骨再生的影响仍不清楚。本研究调查了DXM在体外和体内对成骨的影响。用不同浓度的DXM处理MC3T3-E1前成骨细胞系。进行实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹分析，以评估DXM处理后成骨相关的 runt相关转录因子2（RUNX2）、osterix（OSX）、骨钙素（OCN）、1α型胶原（Col-1α）和碱性磷酸酶（ALP）的表达。将斑马鱼胚胎与可能影响椎骨和颅骨骨化的DXM一起孵育，并使用钙黄绿素染色进行分析。此外，我们使用大鼠颅骨缺损模型，通过微型计算机断层扫描评估DXM对骨再生的影响。结果表明，DXM抑制了MC3T3-E1细胞的细胞外矿化、ALP活性以及成骨标志物RUNX2、OSX、OCN、Col-1α和ALP的表达。DXM抑制了斑马鱼的骨骼骨化，并影响了颅骨缺损大鼠的骨再生。然而，DXM组和对照组再生骨的矿物质密度没有显著差异。本研究表明，DXM对成骨细胞的成骨功能有负面影响，导致骨骼发育和骨再生受损。因此，临床医生应考虑DXM对骨再生的负面影响。
