Hesperetin Prevents Bone Resorption by Inhibiting RANKL-Induced Osteoclastogenesis and Jnk Mediated Irf-3/c-Jun Activation

Abstract

Bone homeostasis and resorption is regulated by the proper activation of osteoclasts, whose stimulation largely depends on the receptor activator of nuclear factor κB ligand (RANKL)-RANK signaling. Herein, for the first time, we showed that interferon regulatory factor (Irf)-3 was intimately involved in RANKL-induced osteoclast formation. In addition, hesperetin (Hes) derived from citrus fruit could inhibit RANKL-induced osteoclast differentiation and maturation among three types of osteoclast precursors with inhibited formation of F-actin rings and resorption pits on bone slices. More importantly, by using SP600125, a selective Jnk inhibitor, we showed that Hes was able to significantly attenuate the Jnk downstream expression of Irf-3 and c-Jun, thereby inactivating NF-κB/MAPK signaling and transcriptional factor NFATc-1, leading to suppression of osteoclast-specific genes, which resulted in impaired osteoclastogenesis and functionality. An ovariectomized (OVX) osteoporosis mouse model demonstrated that Hes could increase trabecular bone volume fractions (BV/TV), trabecular thickness, and trabecular number, whereas it decreased trabecular separation in OVX mice with well-preserved trabecular bone architecture and decreased levels of TRAP-positive osteoclasts. This is further evidenced by the diminished serum expression of bone resorption marker CTX and enhanced production of osteoblastic ALP in vivo. Taken together, these results suggested that Hes could inhibit Jnk-mediated Irf-3/c-Jun activation, thus attenuating RANKL-induced osteoclast formation and function both in vitro and in vivo.

CircRNA_28313/miR-195a/CSF1 axis modulates osteoclast differentiation to affect OVX-induced bone absorption in mice

“…Following a method described before, we established an OVX model in mouse [48] according to the guiding principles of the Animal Care Committee of Second Xiangya Hospital of Central South University to analyze the effects of selected circRNA and miRNA on the osteoporosis in vivo. A total of 32 C57BL/6 mice (8 weeks old, weighted 21.51 ± 0.87 g) were divided into four groups: non-OVX control mice, OVX mice not transfected, OVX mice tail vein injected with Lsh NC (negative control, 100 μl, 1*10 9 TU/ml), and OVX mice injected with (Lsh circRNAs_28313 100 μl, 1*10 9 TU/ml).…”

Tussilagone Inhibits Osteoclastogenesis and Periprosthetic Osteolysis by Suppressing the NF-κB and P38 MAPK Signaling Pathways

“…Primary BMMs were separated from the whole bone marrow of murine femurs and tibias and cultured in complete a-MEM supplemented with 30 ng/ml M-CSF. All cells used in this study were incubated at constant high humidity, 37°C, and 5% CO 2 atmosphere (Zhang et al, 2018). Nonadherent cells were removed before each passage.…”

“…The SYBR Premix Ex Tag kit (TaKaRa Biotechnology) and an ABI 7500 Sequencing Detection System (Applied Biosystems; Foster City, CA, USA) was used in qPCR. PCR conditions were 40 cycles of denaturation at 95°C for 5 s and amplification at 60°C for 34 s (Zhang et al, 2018). The reactions were conducted in triplicate.…”

Tussilagone Inhibits Osteoclastogenesis and Periprosthetic Osteolysis by Suppressing the NF-κb and p38 MAPK Signaling Pathways

“…Quantitative PCR was applied to measure the expression of osteoclast specific genes. We seeded BMMs in 24-well plates at a density of 1×10 5 [26]. The reactions were conducted in triplicate.…”

“…Primary BMMs were separated from the whole bone marrow of murine femurs and tibias and cultured in complete α-MEM with the addition of 30 ng/ml M-CSF. All cells we used in this study were incubated in the incubator with constant high humidity, 37 ℃ and 5% CO2 atmosphere [26]. Non-adherent cells were removed before each passage.…”