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ZCWCC1蛋白在L02肝细胞脂肪变性中的表达变化

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Keywords: 非酒精性脂肪性肝病,脂肪变性,L肝细胞,ZCWCC,泛素-蛋白酶体途径

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Abstract:

目的研究ZCWCC1蛋白在人L02肝细胞脂肪变性中的表达变化并探讨其机制。方法培养中加用500μmol/L的油酸处理L02肝细胞,建立肝细胞脂肪变性体外模型,采用油红染色评价脂质蓄积程度,采用免疫荧光及激光共聚焦显微镜观察ZCWCC1蛋白在L02肝细胞脂肪变性时的表达改变,以Westernblot对其进行半定量评价,并使用赖氨酸蛋白酶抑制剂N-乙基马来酰亚胺(N-Ethylmaleimide,NEM)及蛋白酶体抑制剂MG132对ZCWCC1蛋白表达变化的相关机制进行初步探讨。结果油红染色及细胞甘油三酯含量测定证明L02肝细胞脂肪变性体外模型成功建立,激光共聚焦显微镜观察发现ZCWCC1蛋白主要定位在胞核内,其荧光信号随油酸诱导时间延长而减弱,Westernblot检测结果显示,ZCWCC1表达水平也随油酸诱导时间延长(0、3、6、12、24、48h)而降低[(56283±303)、(56753±539)、(55561±996)、(56257±326)、(48339±712)、(43022±2198),P<0.01]。NEM处理可使ZCWCC1降解增加,MG132处理可逆转ZCWCC1在L02肝细胞脂肪变性中的下降趋势。结论ZCWCC1蛋白在L02肝细胞的脂肪变性过程中表达下降,可能与泛素-蛋白酶体途径有关。

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