2024, Vol. 28 ›› Issue (8): 1149-1154

Urolithin A mediates p38/MAPK pathway to inhibit osteoclast activity

Huang Haoran1, Fan Yinuo2, Wei-Yang Wenxiang1, Jiang Mengyu1, Fang Hanjun1, Wang Haibin1, Chen Zhenqiu1, Liu Yuhao1, Zhou Chi1   

Abstract: BACKGROUND: Overactive osteoclasts disrupt bone homeostasis and play a bad role in the pathological mechanisms of related skeletal diseases, such as osteoporosis, fragility fractures, and osteoarthritis. Studies have confirmed that ellagic acid and ellagtannin have the potential to inhibit osteoclast differentiation. As their natural metabolites, urolithin A has antioxidant, anti-inflammatory, anti-proliferative and anti-cancer effects, but its effect on osteoclast differentiation and its underlying molecular mechanisms remain unclear.
OBJECTIVE: To explore the effect of urolithin A on osteoclast differentiation induced by receptor activator for nuclear factor-κB ligand and its mechanism.
METHODS: Mouse mononuclear macrophage leukemia cells (RAW264.7) that grew stably were cultured in vitro. Toxicity of urolithin A (0, 0.1, 0.5, 1.5, 2.5 μmol/L) to RAW264.7 cells were detected by cytotoxic MTS assay to screen out the safe concentration. Different concentrations of urolithin A were used again to intervene with receptor activator for nuclear factor-κB ligand-induced differentiation of RAW264.7 cells in vitro. Then, tartrate-resistant acid phosphatase staining and F-actin ring and nucleus staining were performed to observe its effect on the formation and function of osteoclasts. Finally, the expressions of urolithin A on upstream and downstream genes and proteins in the MAPK signaling pathway were observed by western blot and RT-qPCR assays.
RESULTS AND CONCLUSION: Urolithin A inhibited osteoclast differentiation and F-actin ring formation in a concentration-dependent manner and 2.5 μmol/L had the strongest inhibitory effect. Urolithin A inhibited the mRNA expression of Nfatc1, Ctsk, Mmp9 and Atp6v0d2 and the protein synthesis of Nfatc1 and Ctsk, related to osteoclast formation and bone resorption. Urolithin A inhibited the activity of osteoclasts by downregulating the phosphorylation of p38 protein to inhibit the mitogen-activated protein kinase signaling pathway.

Key words: urolithin A, osteoclast, MAPK, p38, RANKL