Department of Orthopedic Trauma Surgery, Changzheng Hospital, The Second Military Medical University, Shanghai, China (mainland).
Department of Orthopaedic Trauma Surgery, Changzheng Hospital, The Second Military Medical University, Shanghai, China (mainland).
Med Sci Monit. 2018 Apr 6;24:2045-2052. doi: 10.12659/msm.905703.
BACKGROUND LPS-inhibited osteoblastic differentiation plays an important role in the pathogenesis of osteomyelitis. Thus, searching for drugs that affect LPS-mediated osteoblastic differentiation may be crucial in developing therapies for osteomyelitis. The purpose of this study was to investigate the role and mechanisms of resveratrol, a natural polyphenol present in red wine, on LPS-inhibited osteoblastic differentiation. MATERIAL AND METHODS Cell viability was measured by MMT assay. Mitochondrial ATP levels, membrane potential, and superoxide production were measured to evaluate the effects of LPS and resveratrol on mitochondrial functions in osteoblast-like MC3T3-E1 cells. Osteoblast-related genes, including ALP, OCN, OPN, and RUNX2, were measured by ELISA analysis and RT-PCR in differentiated osteoblast cells treated with LPS and resveratrol. Cellular Sirt1 and PCG-1α levels were measured by Western blot to probe the impact of resveratrol treatment in LPS-stimulated MC3T3-E1 osteoblasts. RESULTS The results showed that LPS caused significant mitochondrial dysfunctions of MC3T3-E1 cells in a dose-dependent manner, which were attenuated by resveratrol. Furthermore, LPS markedly decreased the expression of ALP, OCN, OPN, and RUNX2 in MC3T3-E1 cells cultivated in osteoblast differentiation medium, suggesting that LPS inhibited the osteoblastic differentiation of MC3T3-E1 cells. However, resveratrol obviously alleviated the suppressive impact of LPS on osteoblast differentiation. In addition, resveratrol increased expression of Sirt1 and PGC-1α in MC3T3-E1 cells treated with LPS. CONCLUSIONS Taken together, these results show that resveratrol alleviated the suppression of LPS on osteoblast differentiation by improving, at least in part, mitochondrial function.
脂多糖（LPS）抑制成骨细胞分化在骨髓炎的发病机制中起着重要作用。因此，寻找影响 LPS 介导的成骨细胞分化的药物对于开发骨髓炎的治疗方法可能至关重要。本研究旨在探讨白藜芦醇（一种存在于红酒中的天然多酚）对 LPS 抑制成骨细胞分化的作用及其机制。
通过 MTT 测定法测量细胞活力。通过测量线粒体 ATP 水平、膜电位和超氧化物产生来评估 LPS 和白藜芦醇对成骨样 MC3T3-E1 细胞线粒体功能的影响。用 LPS 和白藜芦醇处理分化的成骨细胞，通过 ELISA 分析和 RT-PCR 测量成骨相关基因，包括碱性磷酸酶（ALP）、骨钙素（OCN）、骨桥蛋白（OPN）和 runt 相关转录因子 2（RUNX2）。通过 Western blot 测量细胞 Sirt1 和过氧化物酶体增殖物激活受体 γ 共激活因子 1α（PGC-1α）的水平，以探讨白藜芦醇处理对 LPS 刺激的 MC3T3-E1 成骨细胞的影响。
结果表明，LPS 以剂量依赖性方式引起 MC3T3-E1 细胞的明显线粒体功能障碍，而白藜芦醇可减轻这种障碍。此外，LPS 显著降低了在成骨细胞分化培养基中培养的 MC3T3-E1 细胞中 ALP、OCN、OPN 和 RUNX2 的表达，表明 LPS 抑制了 MC3T3-E1 细胞的成骨分化。然而，白藜芦醇明显减轻了 LPS 对成骨分化的抑制作用。此外，白藜芦醇增加了 LPS 处理的 MC3T3-E1 细胞中 Sirt1 和 PGC-1α 的表达。
综上所述，这些结果表明，白藜芦醇通过改善线粒体功能（至少部分）缓解了 LPS 对成骨细胞分化的抑制作用。
