BACKGROUND Areca nut has anti-inflammatory, antiparasitic, antihypertensive, and antidepressant properties. The pathological hallmarks of inflammatory joint diseases are an increased number of osteoclasts and impaired differentiation of osteoblasts, which may disrupt the bone remodeling balance and eventually lead to bone loss. PURPOSE The present study assessed the effects of arecoline, the main alkaloid found in areca nut, on osteoclast and osteoblast differentiation and function. METHOD M-CSF/RANKL-stimulated murine bone marrow-derived macrophages (BMMs) were incubated with several concentrations of arecoline, and TRAP staining and pit formation were assessed to monitor osteoclast formation. Quantitative real-time RT-PCR and western blot analyses were used to analyze the expression of osteoclast-associated genes and signaling pathways. The effects of arecoline on bone were investigated in an in vivo mouse model of lipopolysaccharide (LPS)-induced trabecular bone loss after oral administration of arecoline. Alizarin red S staining and assays to measure ALP activity and the transcription level of osteoblast-related genes were used to evaluate the effects of arecoline on osteoblast differentiation and bone mineralization. RESULTS In a dose-dependent manner, arecoline at concentrations of 50-100 μM reduced both the development of TRAP-positive multinucleated osteoclasts and the formation of resorption pits in M-CSF/RANKL-stimulated BMMs. In M-CSF/RANKL-stimulated BMMs, arecoline also suppressed the expression and translocation of c-Fos and NFATcl, and osteoclast differentiated-related genes via interference with the AKT, MAPK, and NF-kB activation pathways. Femur bone loss and microcomputed tomography parameters were recovered by oral administration of arecoline in the mouse LPS-induced bone loss model. Lastly, arecoline increased ALP activity, bone mineralization, and the expression of osteoblast differentiation-related genes, such as ALP and Runx2, in MC3T3-E1 cells. CONCLUSION Our data suggest that arecoline may attenuate or prevent bone loss by suppressing osteoclastogenesis and promoting osteoblastogenesis. These findings provide evidence supporting arecoline's use as a potential therapeutic agent in bone-loss disorders and diseases.

中文翻译：

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槟榔碱在体外抑制RANKL诱导的破骨细胞分化，并在体内减弱LPS诱导的骨损失。

背景技术槟榔具有抗炎，抗寄生虫，抗高血压和抗抑郁的特性。炎性关节疾病的病理学特征是破骨细胞数量增加和成骨细胞分化受损，这可能会破坏骨重塑平衡并最终导致骨质流失。目的本研究评估槟榔碱（槟榔中发现的主要生物碱）对破骨细胞和成骨细胞分化及功能的影响。方法将M-CSF / RANKL刺激的小鼠骨髓巨噬细胞（BMM）与几种浓度的槟榔碱温育，并评估TRAP染色和凹坑形成以监测破骨细胞的形成。实时定量RT-PCR和蛋白质印迹分析用于分析破骨细胞相关基因的表达和信号通路。口服槟榔碱后，在脂多糖（LPS）诱导的小梁骨丢失的体内小鼠模型中研究了槟榔碱对骨骼的影响。茜素红S染色和测定ALP活性以及成骨细胞相关基因的转录水平的方法用于评估槟榔碱对成骨细胞分化和骨矿化的影响。结果以剂量依赖性方式，浓度为50-100μM的槟榔碱可减少TRAP阳性多核破骨细胞的形成以及M-CSF / RANKL刺激的BMMs中吸收凹坑的形成。在M-CSF / RANKL刺激的BMM中，槟榔碱还通过干扰AKT，MAPK和NF-kB激活途径来抑制c-Fos和NFATcl和破骨细胞分化相关基因的表达和转运。通过在小鼠LPS诱导的骨丢失模型中口服槟榔碱可恢复股骨的骨丢失和微计算机断层扫描参数。最后，槟榔碱在MC3T3-E1细胞中增加了ALP活性，骨矿化以及成骨细胞分化相关基因（例如ALP和Runx2）的表达。结论我们的数据表明槟榔碱可以通过抑制破骨细胞生成并促进成骨细胞来减轻或预防骨丢失。这些发现提供了证据支持槟榔碱在骨质疏松症和疾病中作为潜在治疗剂的用途。例如MC3T3-E1单元中的ALP和Runx2。结论我们的数据表明槟榔碱可通过抑制破骨细胞生成并促进成骨细胞来减轻或预防骨丢失。这些发现提供了证据支持槟榔碱在骨质疏松症和疾病中作为潜在治疗剂的用途。例如MC3T3-E1单元中的ALP和Runx2。结论我们的数据表明槟榔碱可通过抑制破骨细胞生成并促进成骨细胞来减轻或预防骨丢失。这些发现提供了证据支持槟榔碱在骨质疏松症和疾病中作为潜在治疗剂的用途。