Marein Prevented LPS-Induced Osteoclastogenesis by Regulating the NF-κB Pathway In Vitro

Abstract

Gram-negative bacterial infection causes many bone diseases such as osteolysis, osteomyelitis and septic arthritis. Lipopolysaccharide (LPS), a bacteria product, played an important role in this process. Drugs that inhibited LPS-induced osteoclastogenesis were urgently needed for the prevention of bone destruction in infective bone diseases. Marein, a major bioactive compound of Coreopsis.tinctoria, which possesses anti-oxidative, anti-in ammatory, anti-hypertensive, anti-hyperlipidemic and anti-diabetic effects. In this study, the effect of marein on RAW264.7 cells was measured by CCK-8 assay; TRAP staining was used to determine osteoclastogenesis; the levels of osteoclast-related genes and NF-κBrelated proteins were analyzed by WB; the levels of pro-in ammatory cytokines were quanti ed by ELISA.Our results showed that marein inhibited LPS-induced osteoclast formation from osteoclast precursor RAW264.7 cells. The effect of marein was related to its inhibitory function on expressions of proin ammatory cytokines and osteoclast-related genes including RANK, TRAF6, MMP-9, CK and CA .Besides, marein treatment could inhibit LPS-induced activation of NF-κB signaling pathway in RAW264.7 cells. Meanwhile, inhibition of NF-κB signaling pathway decreased the formation of osteoclasts and expression of pro-in ammatory cytokines which were LPS-induced. Collectively, marein could prevent LPS-induced osteoclast formation in vitro by regulating NF-κB signaling pathway. These ndings provided evidence that marein might be bene cial as a valuable choice for the prevention and treatment of bacteria-induced bone destruction disease, and gave new insights for understanding its possible mechanism.

Microenvironmental Analysis and Control for Local Cells under Confluent Conditions via a Capillary-Based Microfluidic Device

“…This device is also applicable for analysis of other secreted molecules, and Table S2 summarizes measurable molecules that are secreted from RAW264 cells. As the present device with ELISA detection allowed to detect and quantify TNF-α of 1 pg/mL (60 pM) or larger scale, the molecules in Table S2 are also measurable in their reported concentrations in the 0.1–100 nM scale. − Furthermore, this device produced adjacent stimulated and unstimulated cell groups, which simplified the comparison of the two groups by placing both groups in the same environment.…”

Exploring the pathogenesis of osteomyelitis accompanied by diabetic foot ulcers using microarray data analysis

“…Osteomyelitis is usually caused by infection with Gram-negative bacteria represented by Staphylococcus aureus, and osteoclastogenesis induced by the bacterial product LPS, which promotes bone destruction in osteomyelitis. [ 22 ] In osteomyelitis, LPS stimulates the MAPK signaling pathway, resulting in reduced expression of the anti-inflammatory cytokine IL-10 and reduced quantities of IL-10 recruited. [ 23 ] In DFU, LPS is a significant contributor to insulin resistance and inflammation due to immunopathy and the production of oxidative stress, which directly contribute to the development of DFU.…”