25 December 2019 : Clinical Research
Naringin Protects Against Interleukin 1β (IL-1β)-Induced Human Nucleus Pulposus Cells Degeneration via Downregulation Nuclear Factor kappa B (NF-κB) Pathway and p53 Expression
Gang Gao1AEF, Feng Chang1BD, Ting Zhang1CE, Xinhu Huang1CF, Chen Yu1EF, Zhaolin Hu1BC, Mingming Ji1BE, Yufen Duan2BDG*DOI: 10.12659/MSM.918597
Med Sci Monit 2019; 25:9963-9972
Abstract
BACKGROUND: Low back pain (LBP) is regarded as a frequent disease that causes disability. We aimed to explore the effect of naringin on intervertebral disc degeneration (IDD) in IL-1β-induced human nucleus pulposus (NP) cells and its corresponding molecular mechanisms.
MATERIAL AND METHODS: Human NP cells were identified by toluidine blue and Safranin O staining. Cell viability was determined by MTT assay. The expression levels of matrix metalloproteinases (MMP-3, MMP-13, ADAMTS-4, ADAMTS-5, collagen II, aggrecan), inflammatory genes (tumor necrosis factor [TNF]-α, interleukin [IL]-6), kappa B kinase α (IκBα), p65 and p53 were determined by quantitative real-time polymerase chain reaction (qPCR) and western blotting. Immunofluorescence study was performed to detect the position and expression of p65 protein in IL-1β-induced human NP cells.
RESULTS: Human NP cells were successfully separated from intervertebral disc tissue. We found that naringin could significantly reduce the expressions of matrix metalloproteinases (MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5) and inflammatory genes in IL-1β-stimulated human NP cells, while collagen II and aggrecan were increased at mRNA and protein level. Immunofluorescence showed that naringin pretreatment decreased the p65 protein expression in the nucleus and suppressed the phosphorylation of IκBα and p65.
CONCLUSIONS: These results demonstrated that naringin could attenuate matrix metalloproteinase catabolism and inflammation in IL-1β-treated human nucleus pulposus cells via downregulating NF-κB pathway and p53 expression, suggesting that naringin has the potential to prevent and treat IDD.
Keywords: Cell Nucleus, intervertebral disc degeneration, Cytoprotection, Down-Regulation, Interleukin-6, Matrix Metalloproteinases, nucleus pulposus, Tumor Suppressor Protein p53
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