白藜芦醇对脂多糖诱导的星型胶质细胞炎症损伤的保护作用  

Protective effects of resveratrol on lipopolysaccharide-induced in flammatory toxicity in primary cortical astrocyte cultures

作　　者：刘犇[1] 陈清[1] 郭江[1] 周浩[1] 李劲 俞守义[1] 

机构地区：[1]南方医科大学公共卫生与热带医学学院流行病学系,广东广州510515 [2]解放军第军四五八医院转基因室,广东广州510602

出　　处：《南方医科大学学报》2011年第12期2052-2056,共5页Journal of Southern Medical University

基　　金：广东省自然科学基金重点项目(013075)

摘　　要：目的研究白藜芦醇(RES)对脂多糖诱导的星型胶质细胞炎症因子释放的抑制作用,探索其对中枢神经系统保护作用的可能机制。方法 GFAP免疫组化法鉴定星型胶质细胞的纯度。采用不同浓度的RES与星型胶质细胞共同培养12 h,然后进行LPS损伤24 h,检测不同实验组乳酸脱氢酶(LDH)漏出量,cck-8法检测细胞存活率,Griess法检测NO释放量,Elisa法检测α-肿瘤坏死因子(TNF-α)表达量及Western blot法检测诱导型一氧化氮合酶(iNOS)蛋白的表达水平。结果离体培养的星形胶质细胞的阳性率为(95.49±1.86)%。LPS处理组细胞LDH漏出量升高,存活率降低,同时NO、TNF-α的释放量明显增加及iNOS蛋白的表达水平上调。测试浓度范围(5～50μmol/L)的RES能有效提高细胞存活率及降低LDH漏出量并呈现一定程度的剂量-反应关系。而在抑制NO、TNF-α释放及iNOS表达方面,只有25、50μmol/L的RES处理组有明显抑制作用,5μmol/L RES处理组与LPS处理组相比并无明显变化。结论较高浓度RES能明显抑制LPS诱导的星型胶质细胞炎症介质的释放,改善星型胶质细胞的炎症损伤,这种作用可能与抑制iNOS/NO的表达通路有关。Objective To study the anti-inflammatory effect of resveratrol in primary cortical astrocyte cultures stimulated by lipopolysaccharide（LPS） and explore the underlying mechanism of this protective effect.Methods The astrocytes were cultured in the presence of resveratrol at different concentrations for 12 h followed by stimulation with lipopolysaccharide（LPS） for another 24 h.Lactate dehydrogenas（LDH） leakage volumes were detected,the cytotoxicity of resveratrol was examined using cell counting kit-8（CCK-8）,the release of NO was measured by Griess reaction,and the expression levels of TNF-α and iNOS were measured using ELISA and Western blotting respectively.Results The purity of the astrocytes cultured in vitro was（95.49±1.86）%.LPS treatment increased LDH leakage and reduced the survival rate of the astrocytes,resulting also in significantly increased NO and TNF-α release and iNOS protein expressions.Within the concentration range of 5-50 μmol/L,resveratrol effectively improved the survival rate of the astrocytes and decreased LDH leakage with a dose-response relationship.Only 25 and 50 μmol/L resveratrol produced obvious inhibitory effect on NO and TNF-α release and iNOS expression,while 5 μmol/L resveratrol had no such effects.Conclusion High concentration of resveratrol can inhibit the release of inflammatory mediators and improve the inflammation injury induced by LPS in astrocytes,the mechanism of which may involve the inhibition of iNOS/NO expression pathway.

关 键 词：RES 脂多糖 星形胶质细胞 炎症损伤 

分 类 号：R285.5[医药卫生—中药学]