白藜芦醇通过NF-κB信号通路抑制软骨细胞炎症因子的表达

白藜芦醇通过NF-κB信号通路抑制软骨细胞炎症因子的表达袁昊;曾晖;肖德明;于斐;林健静
【期刊名称】《中国骨与关节外科》
【年(卷),期】2016(009)001
【摘要】背景：炎症因子如IL-1β、TNF-α等可通过一系列级联反应引起关节软
骨破坏和炎症反应，在骨关节炎软骨病变中尤为重要。

白藜芦醇（resveratrol, Res）具有抗炎、抗氧化等作用，研究Res对软骨细胞炎症因子表达的影响可为其预防、治疗骨关节炎提供参考依据。

目的：探讨Res对脂多糖（lipopolysaccharides, LPS）诱导小鼠软骨细胞核因子-κB （nuclear factor-kappa B, NF-κB）活化及炎症因子基因表达的调节及其相关机制。

方法：体外分
离培养小鼠膝关节软骨细胞，Ⅱ型胶原免疫荧光检测进行软骨细胞鉴定。

根据加入不同培养物分为3组：LPS组（1μg/ml LPS）；Res组（100μmol/L
Res+1μg/ml LPS）；空白对照组。

酶联免疫吸附试验（ELISA）法检测各组软骨细胞中NF-κB抑制蛋白α（inhibitor of NF-κB-α, I-κBα）的表达量，细胞免疫
荧光染色观察NF-κB核转位情况，实时定量聚合酶链反应（RT-PCR）测IL-1β、TNF-αmRNA表达量。

结果：荧光显微镜下可见软骨细胞核呈蓝色荧光，胞质呈
红色荧光。

ELISA检测：LPS组的I-κBα表达量低于空白对照组（P＜0.05），Res组明显高于LPS组（P＜0.01）。

细胞免疫荧光染色检测：与空白对照组相比，LPS组中NF-κB发生明显的核转位，而Res组中NF-κB的核转位受到抑制。

RT-PCR检测：LPS组中IL-1β和TNF-αmRNA的表达量与空白对照组相比明显上调，而Res组中IL-1β和TNF-αmRNA的表达量与LPS组相比明显下调，均有统计学差异（P＜0.05）。

结论：Res可通过NF-κB信号通路影响软骨细胞IL-1β、
TNF-αmRNA的表达，进而降低软骨细胞炎症反应，延缓关节软骨细胞退变。

%Background:Inflammatory cytokines such as IL-1 beta, TNF alpha can lead to cartilage destruction and inflam-matory reaction through a series of cascade reaction, especially in cartilage lesions. Resveratrol has anti-inflammatory and antioxidant function. It can provide reference for the prevention and treatment of osteoarthritis by researching the effect of resveratrol on inflammatory cytokines expression of cartilage cells. Objective:To investigate how resveratrol regulates the activation of nuclear factor-Kappa B (NF-κB) and the gene expression of inflammatory cytokines in lipopolysaccharide-in-duced cartilage cells and its related mechanism. Methods: Mouse knee cartilage cells were isolated and cultured in vitro, which were identified by typeⅡcollagen immunofluorescence. The experiment was divided into three groups according to different culture:LPS group (1μg/ml LPS), resveratrol group (100μmol/L
res veratrol+1μg/ml LPS), and blank control group. The expression level of inhibitor of NF-κB-α(I-κBα) were detected by using enzyme-linked immunosorbent assay (ELISA). NF-κB nuclear translocation was observed by immunofluorescence staining. The expression of IL-1β、TNF-αmRNA was measured by real-time quantitative polymerase chain reaction (RT-PCR). Results:The cell nucleus of chondro-cytes was blue and cytoplasm was red under the fluorescent microscope. ELISA detection:the expression of I-κBαin LPS group was si gnificantly lower than that in control group (P<0.05), while the expression of I-κBαin resveratrol group was significantly higher than that in LPS group (P<0.01). Immunofluorescence
staining:the nuclear translocation of NF-κB in LPS group significantly changed compared with the control group, while the nuclear translocation of NF-κB in resveratrol group was inhibited. RT-PCR test:as compared with the control group, the expression of IL-1βand TNF-αmRNA in LPS group were upregulated, while resveratrol group were downregulated compared to LPS group (P<0.05). Conclusions:Resve-ratrol can inhibit the expression of IL-1βand TNF-αmRNA in cartilage cells through NF-κB signal pathway, decrease the inflammatory reaction of chondrocytes, and then delay the degeneration of articular cartilage cells.
【总页数】5页(P75-79)
【作者】袁昊;曾晖;肖德明;于斐;林健静
【作者单位】北京大学深圳医院骨科，广东深圳518036;北京大学深圳医院骨科，广东深圳518036;北京大学深圳医院骨科，广东深圳518036;北京大学深圳医院
骨科，广东深圳518036;北京大学深圳医院骨科，广东深圳518036
【正文语种】中文
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