白藜芦醇抑制脂多糖诱导破骨前体细胞Raw264.7的激活  

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

作　　者：宗一[1] 陈瑾[2] 郭家智[3] 张秀君[3] 张铁军[3] 孙林[4] 

机构地区：[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

出　　处：《解剖学报》2015年第1期51-56,共6页Acta Anatomica Sinica

基　　金：云南省应用基础研究重点项目(2008CC007);云南省中青年学术和技术带头人后备人才培养项目(2009CI033)

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(1μmol/L和5μmol/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(i NOS)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(1μmol/L和5μmol/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i NOS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。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 ceils. 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.

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 

分 类 号：R681[医药卫生—骨科学]