Research Paper Volume 13, Issue 4 pp 5150—5163

Nampt promotes osteogenic differentiation and lipopolysaccharide-induced interleukin-6 secretion in osteoblastic MC3T3-E1 cells

EX527, a selective Sirt1 inhibitor, decreased LPS-induced IL-6 secretion and osteogenic differentiation after LPS treatment. After treatment for 3 days, (A) Real-time PCR analysis showing the mRNA expression of Nampt, Sirt1, and IL-6 normalized to β-actin. (B) Western blot analysis showing the protein expression of Nampt, Sirt1, and p-NF-κB p65. (C) The expression of Sirt1 was measured by fluorescence immunocytochemistry (200×). Red, Sirt1; blue, DAPI. (D) ELISA results showing the IL-6 levels. After EX527 was applied for 7 days, (E) ALP staining and (F) Measurement of ALP activity were decreased. (G) Western blot analysis showing the protein expression of Runx2, Col1a1 and OCN. The data are expressed as the mean ± SD. *, pp

Figure 6. EX527, a selective Sirt1 inhibitor, decreased LPS-induced IL-6 secretion and osteogenic differentiation after LPS treatment. After treatment for 3 days, (A) Real-time PCR analysis showing the mRNA expression of Nampt, Sirt1, and IL-6 normalized to β-actin. (B) Western blot analysis showing the protein expression of Nampt, Sirt1, and p-NF-κB p65. (C) The expression of Sirt1 was measured by fluorescence immunocytochemistry (200×). Red, Sirt1; blue, DAPI. (D) ELISA results showing the IL-6 levels. After EX527 was applied for 7 days, (E) ALP staining and (F) Measurement of ALP activity were decreased. (G) Western blot analysis showing the protein expression of Runx2, Col1a1 and OCN. The data are expressed as the mean ± SD. *, p<0.05; **, p<0.01.