Sanggenon C Stimulates Osteoblastic Proliferation and Differentiation, Inhibits Osteoclastic Resorption, and Ameliorates Prednisone-Induced Osteoporosis in Zebrafish Model

Abstract

Sanggenon C (SC), which is a natural flavonoid found in the stem bark of Cortex Mori, has been discovered to have the antioxidant, anti-inflammatory, and antitumor properties. However, its effect in osteoporosis has not yet been reported. In this research, the effect of SC on the proliferation of MC3T3-E1 cells was evaluated by using the MTT assay. Alkaline phosphatase (ALP) activity and the mRNA expression of Runx2, Collagen I, OPG, and RANKL were examined. TRAP-positive cell counting and bone resorption pits were adopted to observe the effect of SC on the formation and function of osteoclasts. Next, the mRNA level of TRAP, CTSK, NFATc1, and TRAF6 of osteoclasts were measured by real-time qPCR. In addition, the anti-osteoporosis activity of SC in vivo was evaluated in the zebrafish model. Our study indicated that SC exhibited a significant stimulatory effect on MC3T3-E1 cell proliferation at 1 to 10 μM and caused an increase in ALP activity at 0.3 to 10 μM. It could upregulate the expression of Runx2, Collagen I, and increases the OPG/RANKL ratio. Furthermore, SC was found to inhibit the formation and function of osteoclasts, which is demonstrated by a lower number of TRAP-positive multinuclear cells and a fewer area of bone resorption pits compared to the control group. TRAP, CTSK, and NFATc1 were downregulated in 0.3 to 10 μM SC treated groups. In addition, 3 to 10 μM SC also inhibited the expression of TRAF6 mRNA. When prednisone-induced zebrafish was treated with 0.3, 1, 3, and 10 μM SC, higher mineralization of vertebrate column was discovered in a dose-dependent pattern, which suggests that SC could reverse the bone loss of zebrafish caused by prednisone. In summary, these findings indicated that SC has the potential to prevent or treat osteoporosis.

Anti-osteoporosis effects and regulatory mechanism of Lindera aggregata based on network pharmacology and experimental validation

“…68 ctsk encodes a member of the cysteine protease cathepsin family and took part in the bone resorption by the destruction of the collagen matrix of the bone. 65,69 Thus, the two genes ctsk and acp5a of the RA pathway were also detected in this study. The results showed that the mRNA expression of both ctsk and acp5a were greatly decreased in EERL-treated zebrafish compared to the model group.…”

“…64 When RANK binds to RANKL, the complex can stimulate and activate osteoclast differentiation. 65 RANKL/ RANK signaling regulates osteoclast formation, activation, and survival in normal bone remodeling. 66 Thus, RANKL plays a crucial role in bone resorption.…”

Sanggenon C Ameliorates Cerebral Ischemia-Reperfusion Injury by Inhibiting Inflammation and Oxidative Stress through Regulating RhoA-ROCK Signaling

“…It is interesting that an anticancer role of SC in inducing apoptosis of cancer cells had been observed in several previous studies [11,31]. Additionally, SC could also promote the proliferation of osteoblasts while inhibit the formation and function of osteoclasts [21]. The mechanisms underlying when and how SC switches between the pro-and anti-cell apoptosis remain elusive and deserve further investigation.…”

Novel cathepsin K inhibitors block osteoclasts in vitro and increase spinal bone density in zebrafish

“…The use of prednisone can induce bone loss. [36][37][38] In our study, we use prednisone to set up a zebrash bone loss model. Our dosing pilot study showed at the concentration of 1.00 mM, A22 didn't show obvious toxicity to the 30 zerash in the group.…”