CAJ | 학술논문

目的：探讨锌指蛋白A20（zinc finger protein A20）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-G1aN模型组（尾静脉注射D-G1aN 1.8 g/kg）、A20治疗组（尾静脉注射pCAGGS-A20质粒10μg/1 ml NS，8 h后注射D-G1aN 1.8 g/kg）、生理盐水对照组（尾静脉内注射生理盐水1 ml/只）。24 h后，断头取肝组织。ELISA法检测A20、肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）、干扰素-γ（interferon gamma，IFN-γ）蛋白表达水平。qPCR技术检测TNF-α、IFN-γmRNA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（malonic dialdehyd，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-G1aN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-G1aN组与NS组，均P<0.05。A20组的TNF-α以及IFN-γmRNA和蛋白质表达水平均显著低于D-G1aN组，均P<0.05。A20组小鼠肝脏总SOD活性显著高于D-G1aN组，P<0.05，MDA、MPO活性显著低于D-G1aN组，均P<0.05。A20组小鼠肝组织病理学改变显著轻于D-G1aN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。
목적：탐토자지단백A20（zinc finger protein A20）대D-반유당알（D-galactosamine，D-GlaN）인기적급성간손상적보호작용。방법45지C57BL/6수궤균분위삼조：D-G1aN모형조（미정맥주사D-G1aN 1.8 g/kg）、A20치료조（미정맥주사pCAGGS-A20질립10μg/1 ml NS，8 h후주사D-G1aN 1.8 g/kg）、생리염수대조조（미정맥내주사생리염수1 ml/지）。24 h후，단두취간조직。ELISA법검측A20、종류배사인자-α（tumor necrosis factor-α，TNF-α）、간우소-γ（interferon gamma，IFN-γ）단백표체수평。qPCR기술검측TNF-α、IFN-γmRNA표체수평。ELISA법검측간조직내초양화물기화매（superoxide-dimutase，SOD）、병이철（malonic dialdehyd，MDA）、수과양화물매（myeloperoxidase ，MPO）활성적변화。응용HE염색기술관찰간병리손상정도。결과D-G1aN조적A20단백질함량현저고우NS조，P<0.05，A20조적A20단백질함량우현저고우D-G1aN조여NS조，균P<0.05。A20조적TNF-α이급IFN-γmRNA화단백질표체수평균현저저우D-G1aN조，균P<0.05。A20조소서간장총SOD활성현저고우D-G1aN조，P<0.05，MDA、MPO활성현저저우D-G1aN조，균P<0.05。A20조소서간조직병이학개변현저경우D-G1aN조。결론A20구유항양화손상적효응，A20전기인치료능구유효억제D-GalN인기적염증반응。
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-G1aN 1.8 g/kg through caudal vein, A20 group injected with pCAGGS-A20 plasmid 10μg/1 ml NS, and then D-G1aN 1.8 g/kg 8 h later through caudal vein, and NS group injected 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-α(TNF-α), interferon-γ(IFN-γ) were detected by ELISA, the mRNA expression of TNF-αand IFN-γwere 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-G1aN 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-G1aN group,P<0.05. The mRNA and protein expression level TNF-αand IFN-γin 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-G1aN 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-G1aN 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-G1aN .