锌指蛋白A20对D-半乳糖胺诱导小鼠急性肝损伤的保护作用

Protective effect of zinc finger protein A20 on D-galactosamine-induced acute hepatic injury in mice

目的探讨锌指蛋白A20（zincfingerproteinA20）对D-半乳糖胺（D-galactosamine，D-GlaN）引起的急性肝损伤的保护作用。方法45只C57BL／6随机均分为三组：D-GlaN模型组（尾静脉注射D—GlaN1．8g／k力、A20治疗组（胃邑嫠H明℃沿射pCAGGS—A20质粒10μg／imlNS，8h斥；注射D—GlaN1．8班亩、生理盐水对照组（尾静脉内注射生理盐水1ml／只）。24h后，断头取肝组织。ELISA法检测A20、肿瘤坏死因子-α（tumornecrosisfactor-α，TNF-α、干扰素一1（interferongamma，IFN-γ）蛋白表达水平。qPCR技术检测TNF-α、IFN-γ／mRNA表达水平。ELISA法检测肝组织内超氧化物岐化酶（superoxide—dimutase，SOD）、丙二醛（malonicdialdehyd，MDA）、髓过氧化物酶（myeloperoxidase，MPO）活性的变化。应用HE染色技术观察肝病理损伤程度。结果D—GlaN组的A20蛋白质含量显著高于NS组，P〈0．05，A20组的A20蛋白质含量又显著高于D-GlaN组与Ns组，均P〈0．05。A20组的TNF—α以及IFN-γ／mRNA和蛋白质表达水平均显著低于D-GlaN组，均P〈0．05。A20组小鼠肝脏总SOD活性显著高于D—-GlaN组，P〈0．05，MDA、MPO活性显著低于D-GlaN组，均P〈O．05。A20组小鼠肝组织病理学改变显著轻于D—GlaN组。结论A20具有抗氧化损伤的效应，A20转基因治疗能够有效抑制D—GalN引起的炎症反应。

Objective To explore the protective effect of zinc finger protein A20 on acute hepatic injury in mice induced by D-galactosamine. Methods Forty-five C57BL/6 mice were divided into three groups： D-galactosamine （D-GlaN） group injected with D-GlaN 1.8 g/kg through caudal vein, A20 group injected with pCAGGS-A20 plasmid 10 p,g/1 ml NS, and then D-GlaN 1.8 g/kg 8 h later through caudal vein, and NS group injectSd with NS 1 ml. Twenty-four hours later the mice were killed with their livers collected. The protein expression levels of A20, tumor necrosis factor-c~ （TNF-c~）, interferon-~/（IFN-~/） were detected by ELISA, the mRNA expression of TNF-c~ and IFN-~/were detected by reverse transcription polymerase chain reaction. The levels of superoxide-dimutase （SOD）, malonic dialdehyd （MDA）, and myeloperoxidase （MPO） were examined. The liver tissues underwent pathological examination with HE staining. Results The A20 protein level of the D-GlaN group was significantly higher than that of the NS group, P 〈0.05, and the A20 protein level of the A20 group was significantly higher than that of the D-GlaN group, P 〈0.05. The mRNA and protein expression level TNF-ot and IFN-~/in the liver tissue of the A20 group were both significantly lower than those of the D-GlaN group , both P 〈0.05. The activity level of SOD in the liver tissue of the A20 group was significantly higher than that of the D-GlaN group, P 〈0.05, and the activity levels of MDA and MPO in the liver tissue of the A20 group were both lower than those of the D-GlaN group , both P 〈0.05. The pathological changes in the liver tissue of the A20 group were significantly milder than those of the D-GlaN group. Conclusion A20 has the function against oxidative damage. A20 trans-gene therapy effectively inhibit the inflammatory response induced by D-GlaN.