Department of Pharmacology, Institute of Materia Medica, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100050, China.
Acta Pharmacol Sin. 2010 Apr;31(4):470-5. doi: 10.1038/aps.2009.202. Epub 2010 Mar 8.
To study the effects and mechanism of aromatic aminoketone (SY0916) on bone destruction in vitro.
MC3T3-E1 cells and bone marrow cells were co-cultured to obtain purified osteoclasts. The proliferation of osteoclast-like cells (OCLs) was determined by MTT assay. The number of osteoclasts was measured by tartrate-resistant acid phosphatase (TRAP) staining. The functioning of osteoclasts was determined by measuring the area of bone resorption pits on bone slices. MMP-9 secretion by osteoclasts was measured by an ELISA kit. Osteoclast apoptosis was detected by flow cytometry using an AnnexinV-FITC kit. Gene expression of RANK and MMP-9 in osteoclasts as well as RANKL and OPG in MC3T3-E1 cells was determined by real-time PCR.
SY0916 significantly inhibited the proliferation of OCLs, decreased both the total and average area of bone resorption pits, and dramatically inhibited the number of osteoclasts between concentrations of 0.01 and 10 micromol/L. Furthermore, SY0916 reversed IL-1 beta-mediated inhibition of osteoclast apoptosis and shortened osteoclast lifespan. In addition, SY0916 significantly inhibited the mRNA expression of RANK, RANKL, OPG, and MMP-9. However, the inhibition of OPG was weaker than that of RANKL. Accordingly, the ratio of RANKL to OPG mRNA expression in MC3T3-E1 cells was significantly decreased by SY0916. Meanwhile, the expression of MMP-9 protein in osteoclasts was inhibited by SY0916 between 0.01 and 10 micromol/L.
SY0916 prevents osteoclastic bone destruction by inhibiting the proliferation and function of osteoclasts. The underlying mechanism for this effect involves the regulation of the RANKL-OPG-RANK axis, which determines the direction of bone metabolism.
研究芳香族氨基酮（SY0916）对体外骨破坏的影响及其机制。
MC3T3-E1 细胞与骨髓细胞共培养，获得纯化的破骨细胞。MTT 法检测破骨样细胞（OCL）的增殖。抗酒石酸酸性磷酸酶（TRAP）染色法测定破骨细胞数量。骨切片上骨吸收陷窝面积测定法测定破骨细胞功能。酶联免疫吸附试验（ELISA）试剂盒测定破骨细胞 MMP-9 的分泌。AnnexinV-FITC 试剂盒通过流式细胞术检测破骨细胞凋亡。实时 PCR 检测破骨细胞中 RANK 和 MMP-9 的基因表达以及 MC3T3-E1 细胞中 RANKL 和 OPG 的基因表达。
SY0916 显著抑制 OCL 的增殖，在 0.01 至 10 微摩尔/升浓度范围内，同时减少总骨吸收陷窝面积和平均骨吸收陷窝面积，并显著抑制破骨细胞数量。此外，SY0916 逆转了 IL-1β对破骨细胞凋亡的抑制作用，缩短了破骨细胞的寿命。此外，SY0916 显著抑制 RANK、RANKL、OPG 和 MMP-9 的 mRNA 表达。然而，OPG 的抑制作用弱于 RANKL。因此，SY0916 显著降低了 MC3T3-E1 细胞中 RANKL 与 OPG mRNA 表达的比率。同时，SY0916 在 0.01 至 10 微摩尔/升浓度范围内抑制了破骨细胞中 MMP-9 蛋白的表达。
SY0916 通过抑制破骨细胞的增殖和功能来预防破骨细胞性骨破坏。这种作用的潜在机制涉及调节 RANKL-OPG-RANK 轴，该轴决定了骨代谢的方向。
