ZCWCC1蛋白在L02肝细胞脂肪变性中的表达变化

发布时间：2018-11-19 11:12

【摘要】：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Western blot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,Western blot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、48 h)而降低[(56 283±303)、(56 753±539)、(55 561±996)、(56 257±326)、(48 339±712)、(43 022±2 198),P0.01]。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。
[Abstract]:Objective to investigate the expression and mechanism of ZCWCC1 protein in human L02 hepatocyte fatty degeneration. Methods L02 hepatocytes were treated with 500 渭 mol/L oleic acid in vitro. The fatty degeneration model of hepatocytes was established in vitro. The degree of lipid accumulation was evaluated by oil red staining. Immunofluorescence and laser confocal microscopy were used to observe the expression of ZCWCC1 protein in L02 hepatocytes during steatosis. Western blot was used to evaluate the expression of ZCWCC1 protein. N-ethylmaleimide (N-ethylmaleimide), a lysine protease inhibitor, was used to evaluate the expression of N-ethylmaleimide (N-ethylmaleimide). NEM) and proteasome inhibitor MG132 to investigate the mechanism of ZCWCC1 protein expression. Results the oil red staining and the determination of triglyceride content showed that L02 hepatocyte fatty degeneration model was successfully established in vitro. Laser confocal microscopy showed that ZCWCC1 protein was mainly located in the nucleus. The results of, Western blot analysis showed that the level of ZCWCC1 expression decreased with the prolongation of oleic acid induction time (0 ~ 3 ~ (3), () [(56 283 卤303), (56 753 卤539), (55 561 卤996), (56 257 卤326), (48 339 卤712]. (43 022 卤2 198), P0.01]. NEM treatment could increase ZCWCC1 degradation, MG132 treatment could reverse the decreasing trend of ZCWCC1 in L02 hepatocyte steatosis. Conclusion the decrease of ZCWCC1 protein expression in L02 hepatocytes may be related to the ubiquitin proteasome pathway.
【作者单位】：第三军医大学大坪医院野战外科研究所消化内科;解放军75210部队医院外科;
【基金】：国家自然科学基金(81170382)~~
【分类号】：R575.5

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