Résumé
Objective To observe the effect of Juncus effuses on osteoclasts differentiation from bone marrow macrophages (BMMs) induced by receptor activator for nuclear factor-κ B ligand (RANKL), and its mechanism thereof. Methods BMMs were isolated from whole bone marrow of 8-week-old C57/BL6 mice, and CCK-8 was used to detect the effect of Juncus on BMMs cell proliferation. Tartrate resistant acid phosphatase (TRAP) staining was used to show that 50 μg/L RANKL and 30 μg/L macrophage colony stimulating factor (M-CSF) stimulated the BMMs differentiation into osteoclasts, but the process was inhibited by Juncus (0, 6.25, 12.5 and 25 μmol/L). RT-PCR was used to detect the expressions of osteoclast-specific genes including calcitonin receptor (CTR), vacuolated H+triphosphate transporter -d2 (V-ATPase-d2) and -a3 (V-ATPase-a3), activated T nuclear factor 1 (NFATC1) and C-FOS. Results There was no inhibition in the proliferation of BMMs cells treated with Juncus less than 12.5 μmol/L detected by CCK-8. The 50 μg/L RANKL can induce BMMs differentiated into positive multinuclear giant cells detected by TRAP staining, but Juncus significantly inhibited osteoclast formation with a concentration dependence. The results of RT-PCR experiment showed that Juncus inhibited the expression of specific genes in osteoclast differentiation in concentration-dependent manner. Conclusion Juncus can inhibit osteoclast formation in concentration-dependent manner, resulting from the inhibitory effect on osteoclast specific gene expression.
Résumé
Objective: To rapidly recognize and identify phenanthrenes in Juncus effusus and J. setchuensis. Methods: HPLC-ESI-MS method was used to investigate the ESI-MS characteristics of those phenanthrenes, to screen, and to identify phenanthrenes in J. effusus and J. setchuensis. Results: Under negative mode, phenanthrenes were inclined to loss two hydrogens, which presumably caused by the cyclization of vinyl and aromatic ring; Furthermore, the loss of C=O could be also observed. Fragmentation pathway of phenanthrenes in ESI as a negative ion mode was summarized. Twenty-one phenanthrenes were rapidly detected from the extracts of J. effusus and J. setchuensis. Among them, six compounds were accurately identified by comparison with reference substances; The structures of five compounds were inferred according to their MS spectrum and polarity. Conclusion: HPLC-ESI-MS provides not only a new technique for the rapid identification of phenanthrenes from complex matrix, but also an effective method for the target separation of constituents in Chinese materia medica.