姜黄素对氧化应激中肝星状细胞活化及细胞外基质分泌的影响

Influence of curcumin on hepatic stellate cells activation and extracellular matrix secretion in oxidative stress

目的观察姜黄素对氧化应激中大鼠肝星状细胞系HSC-T6活化及细胞外基质分泌的影响。方法将HSC-T6分成对照组、模型组、干预组。对照组正常培养,模型组加入100 mU/mL葡萄糖氧化酶(glucose oxidase,GO)干预2 h,制备细胞氧化应激模型,干预组分别加入15μmol/L和30μmol/L姜黄素分别干预3 h,然后给予100 mU/mL GO干预2 h。免疫荧光法观察HSC-T6中desmin和α-SMA的表达,分光光度法检测细胞丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)水平,放射免疫法检测细胞内透明质酸(hyaluronic acid,HA),层粘连蛋白(laminin,LN),Ⅲ型前胶原(procollagenⅢ,PCⅢ),Ⅳ型胶原(collagenⅣ,CⅣ)水平的变化。结果模型组MDA较对照组显著升高(P<0.01),姜黄素15μmol/L干预组MDA较模型组有所降低(P<0.05),姜黄素30μmol/L干预组MDA较模型组明显降低(P<0.01);模型组GSH较对照组显著升高(P<0.01),姜黄素干预组GSH均较模型组显著升高(P<0.01),姜黄素30μmol/L的作用优于15μmol/L;对照组中α-SMA无明显表达,模型组中α-SMA大量表达,干预组中α-SMA的表达明显减少,其中姜黄素30μmol/L的作用优于15μmol/L;模型组HA、LN、PCⅢ、CⅣ水平较对照组显著升高(P<0.01),姜黄素15μmol/L干预组细胞内HA、LN、PCⅢ、CⅣ水平较模型组有所降低(P<0.05),姜黄素30μmol/L干预组HA、LN、PCⅢ、CⅣ水平较模型组明显降低(P<0.01)。结论姜黄素在低浓度内能够呈浓度依赖性地降低星状细胞内氧化应激水平,抑制星状细胞活化进而减轻细胞纤维化。

Objective To investigate the influence of curcumin on hepatic stellate cells（HSC-T6） activation and extracellular matrix secretion in oxidative stress.Methods HSC-T6s were divided into control group,model group and curcumin intervention group.HSC-T6s in control group were only incubated with high glucose DMEM,HSC-T6s in model group were treated with 100 mU/mL GO for 2 h,HSC-T6s in curcumin intervention group were treated with curcumin 30 μmol/L and 15 μmol/L for 3 h respectively and treated with 100 mU/mL GO for 2 h subsequently.Then the HSC-T6s were analyzed to observe the expression of desmin and α-SMA by immunofluorescence.The levels of MDA and GSH were measured by spectrophotometric method.The levels of intracellular HA,LN,PCⅢ,CⅣ were determined by radioimmunity method.Results Compared with control group,the level of MDA increased significantly in model group（P0.01）,and the level of MDA in 15 μmol/L curcumin intervention group decreased compared with model group（P0.05）,while the level of MDA in 30 μmol/L curcumin intervention group significantly decreased compared with model group（P0.01）.The level of GSH in model group was higher than that in control group（P0.01）.However,the level of GSH increased in curcumin intervention group compared with model group（P0.01）,the effect of 30 μmol/L curcumin was better than that of 15 μmol/L curcumin.The expression of α-SMA in control group was negative,while there was abundant expression in model group.The expression of α-SMA decreased compared with model group,and the effect of 30 μmol/L curcumin was better than that of 15 μmol/L curcumin.The levels of HA,LN,PCⅢ,CⅣ in model group upgraded apparently compared with control group（P0.01）,and the levels in 15 μmol/L curcumin group were lower than those in model group（P0.05）,while the levels in 30 μmol/L curcumin group were significantly lower than those in model group（P0.01）.Conclusion Curcumin can dilute the level of oxidation,depress HSC activation and lessen fibrosis.