Objective

The helioxanthin derivative 4-(4-methoxyphenyl)thieno[2,3-b:5,4-c′]dipyridine-2-carboxamide (TH) is a low-molecular-weight compound that was identified through screening for osteogenic compounds that enhance the activity of mouse preosteoblastic MC3T3-E1 cells. In the present study, we found that TH suppressed osteoclast differentiation.

Methods

Using the hematopoietic stem cells of ddY mice, TH was added to the culture in the experimental group, and the number of osteoclasts was measured with rhodamine phalloidin staining and TRAP staining. In osteo assay, bone resorption area was compared by the von Kossa staining.

Results

Specifically, TH inhibited the cyclic guanosine monophosphate (cGMP)-degrading activity of phosphodiesterase (PDE), promoted nitric oxide (NO) production, and dose-dependently suppressed osteoclast differentiation in an osteoclast formation culture of mouse bone marrow cells. The NO-competitive guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) attenuated the suppressive activity of TH on osteoclast differentiation. Conclusion: Given the previously reported suppressive action of cGMP on osteoclastogenesis, our data suggest that TH negatively impacts osteoclast differentiation at least to some extent by stimulating NO production and inhibiting PDE activity, both of which lead to the upregulation of intracellular cGMP. This study supports the potential use of TH as a novel antiosteoporotic reagent that not only stimulates bone formation but also inhibits bone resorption.

中文翻译：

---

一种成骨的螺旋黄质衍生物抑制骨吸收破骨细胞的形成。

客观的

helioxanthin 衍生物 4-(4-methoxyphenyl)thieno[2,3-b:5,4-c']dipyridine-2-carboxamide (TH) 是一种低分子量化合物，通过筛选成骨化合物确定增强小鼠前成骨细胞 MC3T3-E1 细胞的活性。在本研究中，我们发现 TH 抑制破骨细胞分化。

方法

使用ddY小鼠的造血干细胞，在实验组培养物中加入TH，用罗丹明鬼笔环肽染色和TRAP染色测定破骨细胞的数量。在骨测定中，通过冯科萨染色比较骨吸收面积。

结果

具体来说，在小鼠骨髓细胞的破骨细胞形成培养物中，TH 抑制了磷酸二酯酶 (PDE) 的环磷酸鸟苷 (cGMP) 降解活性，促进一氧化氮 (NO) 的产生，并且剂量依赖性地抑制破骨细胞分化。NO 竞争性鸟苷酸环化酶抑制剂 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) 减弱了 TH 对破骨细胞分化的抑制活性。结论：鉴于先前报道的 cGMP 对破骨细胞生成的抑制作用，我们的数据表明 TH 通过刺激 NO 产生和抑制 PDE 活性至少在一定程度上对破骨细胞分化产生负面影响，这两者都会导致细胞内 cGMP 的上调。