解剖学报

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

• 细胞和分子生物学 • 上一篇    下一篇

白藜芦醇抑制脂多糖诱导破骨前体细胞Raw 264.7的激活

宗一1 陈瑾2 郭家智3 张秀君3 张铁军3 孙林4*   

  1. 1. 浙江大学医学院附属第四医院放射科，浙江 义乌 322000; 2. 义乌市中心医院血液科，浙江 义乌 322000; 3. 昆明医学院基础医学院人体解剖学与组织学胚胎学系，昆明 650500; 4. 昆明医学院第二附属医院心内科，昆明 650101
  • 收稿日期:2014-06-23 修回日期:2014-09-03 出版日期:2015-02-06 发布日期:2015-02-06
  • 通讯作者: 宗一 E-mail:yiwuzongyi2001@gmail.com
  • 基金资助:

    云南省应用基础研究重点项目;云南省中青年学术和技术带头人后备人才培养项目

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

ZONG Yi1 CHEN Jin2 GUO Jia-zhi3 ZHANG Xiu-jun3 ZHANG Tie-jun3  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

摘要：

目的 探讨白藜芦醇 (Res) 对脂多糖 (LPS) 诱导的破骨前体细胞 Raw 264.7 细胞系释放炎性细胞因子的抑制作用。方法 采用 LPS 刺激 Raw 264.7 细胞构建炎症模型，采用抗酒石酸酸性磷酸酶 (TRAP)染色鉴定细胞，采用MTT 检测 Res 对 Raw 264.7 细胞的毒性影响，免疫荧光双标以及反转录聚合酶链反应(RT-PCR) 方法检测不同浓度 Res (1 μmol/L和 5 μmol/L) 对细胞炎性因子：肿瘤坏死因子-α (TNF-α) 和白细胞介素-1β (IL-1β) 与细胞炎性蛋白酶：诱导型一氧化氮合酶 (iNOS) 和环氧合酶-2 (COX-2)、炎性信号蛋白核因子-κB (NF-κB) 蛋白与 mRNA 的表达变化。 结果 不同浓度的 Res (1 μmol/L和5 μmol/L)在翻译水平和转录水平上明显抑制了 LPS 诱导的细胞炎性蛋白酶 iNOS 和 COX-2表达，同时了抑制细胞炎性因子 IL-1β 与炎性信号蛋白 NF-κB 的上调。 结论 Res 可能通过 NF-κB 调控 LPS 诱导的 Raw 264.7 细胞炎性细胞因子的释放进而抑制破骨前体细胞的激活，进而具有抗骨质疏松的作用。

关键词: 白藜芦醇, 脂多糖, Raw 264.7细胞, 炎性因子, 反转录聚合酶链反应, 免疫荧光双标, 小鼠

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