AAS ›› 2015, Vol. 46 ›› Issue (1): 51-56.doi: 10.16098/j.issn.0529-1356.2015.01.009

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Resveratrol inhibits lipopolysaccharide-induced activation of osteoclast precursor Raw 264.7 cells

ZONG Yi1 CHEN Jin2 GUO Jia-zhi3 ZHANG Xiu-jun3 ZHANG Tie-jun SUN Lin 4*   

  1. 1. Department of Radiology, the Fourth Affiliated Hospital of Zhejiang University School of Medicine, Zhejiang Yiwu 322000, China; 2. Department of Hematology, the Central Hospital of Yiwu City, Zhejiang Yiwu 322000, China; 3. Department of Anatomy, Kunming Medical University, Kunming 650500, China; 4. Department of Cardiology, the Second Affiliated Hospital of Kunming Medical University, Kunming 650101, China
  • Received:2014-06-23 Revised:2014-09-03 Online:2015-02-06 Published:2015-02-06

Abstract:

Objective To investigate the inhibiting effects of resveratrol (Res) on the expression of potentially inflammatory cytokines by cultured osteoclast precursor Raw 264.7 cells stimulated with lipopolysaccharide (LPS). Methods Inflammatory cell model was established by LPS-stimulated Raw 264.7 cells. The Raw 264.7 cell identification test was measured by tartrate resistant acid phosphatase (TRAP) staining. The cells were treated with Res (1 μmol/L and 5 μmol/L) prior to LPS (5 mg/L) exposure. The effects on the mRNA and protein levels of inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and inflammatory cytokines, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β), inflammatory signaling proteins nuclear factor-κB (NF-κB) were analysed by reverse transcription-polymerase chain reaction (RT-PCR) and double-immunofluorescence labeling assay. The effects of Res on cytotoxicity determination of Raw 264.7 cells were measured by MTT assay. Results LPS-induced iNOS, COX-2 and NF-κB protein and mRNA expression levels were significantly decreased by Res. Res had an effect on the expression of TNF-α, IL-1β through transcriptional and translational inhibition. Conclusion The inhibitory effects of Res on LPS-induced inflammatory mediators in osteoclast precursor cells exert functions on its anti-osteoporosis through NF-κB.

Key words: Resveratrol, Lipopolysaccharide, Raw 264.7 cells, Inflammatory cytokines, RT-PCR, Double-immunofluorescence labeling, Mouselabeling