Zoledronic acid inhibits lipopolysaccharide-induced osteoclast differentiation by regulating NLRP3 signaling pathway

doi:10.12307/2023.425

Abstract

Abstract: BACKGROUND: Zoledronic acid can inhibit bone resorption, but whether it can inhibit inflammatory bone disease and its related mechanisms are completely unclear.
OBJECTIVE: To investigate the effect of zoledronic acid on the proliferation and differentiation of osteoclasts induced by lipopolysaccharide.
METHODS: RAW264.7 cells were treated with lipopolysaccharide for osteoclastic induction and dyed with tartrate-resistant acid phosphatase and F-actin. RAW264.7 cells were cultured in vitro and divided into blank control group (no intervention), control group (treated with lipopolysaccharide), and 0.1, 1, 5 μmol/L zoledronic acid groups (lipopolysaccharide+0.1, 1, 5 μmol/L zoledronic acid). After 6 hours of culture, the level of tumor necrosis factor α in the supernatant was detected by ELISA, and the protein expression levels of NLRP3, caspase-1, interleukin-1β, cleaved-caspase-1 and tumor necrosis factor α were detected by western blot assay. On the 5th day of culture, the formation of osteoclasts was observed by tartrate-resistant acid phosphatase staining, and the expression of actin rings in osteoclasts was observed by F-actin staining.
RESULTS AND CONCLUSION: (1) Lipopolysaccharide could induce RAW264.7 cells to differentiate into osteoclasts. (2) The level of tumor necrosis factor α was higher in the control group than the blank control group (P < 0.05) and lower in the 0.1 and 5 μmol/L zoledronic acid groups than the control group (P < 0.05). Results from tartrate-resistant acid phosphatase and F-actin staining indicated that 1 and 5 μmol/L zoledronic acid could inhibit lipopolysaccharide-induced differentiation of RAW264.7 cells into osteoclasts. (3) Compared with the control group, 0.1, 1, and 5 μmol/L zoledronic acid could inhibit the protein expression of NLRP3, caspase-1, interleukin-1β, and tumor necrosis factor α (P < 0.05), while 1 and 5 μmol/L zoledronic acid could inhibit the protein expression of cleaved-caspase-1 (P < 0.05). (4) To conclude, zoledronic acid can inhibit osteoclast formation induced by lipopolysaccharide, which acts possibly through regulating the NLRP3 signaling pathway.

Key words: zoledronic acid, lipopolysaccharide, osteoclast, RAW264.7 cell, NLRP3 signaling pathway, tartrate-resistant acid phosphatase, inflammatory bone disease, bone infection

CLC Number:

Liu Guanjuan, Song Na, Huo Hua, Luo Shanshan, Cheng Yuting, Xiong Yue, Hong Wei, Liao Jian. Zoledronic acid inhibits lipopolysaccharide-induced osteoclast differentiation by regulating NLRP3 signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(29): 4677-4683.

Figures/Tables 9

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