N‑methyl pyrrolidone promotes ankle fracture healing by inhibiting inflammation via suppression of the mitogen‑activated protein kinase signaling pathway

Abstract

Abstract. N-methyl pyrrolidone (NMP), a small bioactive molecule, has the potential to stimulate bone formation and inhibit osteoclast differentiation. The aim of the present study was to investigate the effect of NMP on the inflammatory response and underlying molecular mechanisms in MG-63 cells. The mRNA and protein expression of cytokines from peripheral blood in children with or without ankle fracture were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and ELISA, respectively. MG-63 cells were pre-treated with/without NMP and stimulated with 1 µM bradykinin (BK). The production of cytokines from MG-63 cells was assessed by western blotting and RT-qPCR. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA and protein were measured using western blotting and/or RT-qPCR. Western blotting was used to examine the activation level of mitogen activated protein kinase. Compared with healthy children, levels of tumor necrosis factor (TNF-α), interleukin (IL)-1β and IL-6 mRNA and protein were upregulated in children with ankle fracture. NMP treatment did not induce cytotoxicity in MG-63 cells. The BK-induced upregulation of TNF-α, IL-1β, IL-6, iNOS and COX-2 mRNA and protein was reversed in a dose-dependent manner by NMP. Furthermore, NMP downregulated the activation of c-Jun NH2-terminal kinase and p38 pathways, but not the extracellular signal-related kinase pathway. Therefore, the results of the current study demonstrate that NMP inhibits inflammation dependent on the mitogen-activated protein kinase pathway in MG-63 cells, indicating that it may be beneficial in the healing of fractures.

Determination of immune factor levels in serum and local hematoma samples of osteoporotic fracture patients and clinical study of the effect of active vitamin D3 treatment on immune factor levels

“…Some studies have found through animal experiments that the expression of IL-1β protein in the joint cavity of mice is significantly reduced after blocking TNF-α, suggesting that TNF-α is related to the production of IL-1β protein [ 19 ]. Similar conclusions have been reached in cell experiments, where MSU crystals can more effectively promote the synthesis of IL-1β precursor and IL-18 in medium supplemented with TNF-α [ 20 ]. TNF-α also has dual effects on fracture: on the one hand, it reduces new bone formation, and on the other hand, it enhances bone resorption, improves OC activity, and reduces cortical bone quality.…”

Determination of immune factor levels in serum and local hematoma samples of osteoporotic fracture patients and clinical study of the effect of active vitamin D3 treatment on immune factor levels

“…Some studies have found through animal experiments that the expression of IL-1β protein in the joint cavity of mice is signi cantly reduced after blocking TNF-α, suggesting that TNF-α is related to the production of IL-1β protein [10] . Similar conclusions have been reached in cell experiments, where MSU crystals can more effectively promote the synthesis of IL-1β precursor and IL-18 in medium supplemented with TNF-α [11] . TNF-α also has dual effects on fracture: on the one hand, it reduces new bone formation, and on the other hand, it enhances bone resorption, improves OC activity, and reduces cortical bone quality.…”

Cervus and cucumis peptides combined umbilical cord mesenchymal stem cells therapy for rheumatoid arthritis

“…It contains bone-induced polypeptide biological factors, such as transforming growth factor -beta (TGF- beta), [ 14 ] which can promote the synthesis of type I collagen and osteoporosis. [ 14 ] They inhibit the inflammatory response, reduce the content of tumor necrosis factor-alpha (TNF-alpha) in serum, [ 15 ] and assist macrophage-derived cytokines to play a role in tissue repair. [ 16 ] According to the pathogenesis of RA in traditional Chinese and western medicine, UC-MSCs are pre-stimulated by the polypeptide of LG to promote the superposition of kidney-nourishing, bone strengthening, blood-activating, and collateral-opening, and also reduce the release of pro-inflammatory factors.…”