[1]曹洁,杨朝霞,沈薇,等.内质网应激在软脂酸钠诱导的脂肪变性L02肝细胞凋亡中的作用[J].陆军军医大学学报(原第三军医大学学报),2011,33(18):1935-1938.
 Cao Jie,Yang Zhaoxia,Shen Wei,et al.Role of endoplasmic reticulum stress in palmitate sodium-induced apoptosis in steatotic L02 hepatocytes[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(18):1935-1938.
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内质网应激在软脂酸钠诱导的脂肪变性L02肝细胞凋亡中的作用(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
33卷
期数:
2011年第18期
页码:
1935-1938
栏目:
论著
出版日期:
2011-09-30

文章信息/Info

Title:
Role of endoplasmic reticulum stress in palmitate sodium-induced apoptosis in steatotic L02 hepatocytes
作者:
曹洁杨朝霞沈薇姚隆
重庆医科大学附属第二医院消化科;江津区中心医院重症医学科
Author(s):
Cao Jie Yang Zhaoxia Shen Wei Yao Long
Department of Gastroenterology, Second Affiliated Hospital, Chongqing Medical University,Chongqing, 400010; Department of Intensive-care Medicine, Central Hospital of Jiangjin District, Chongqing, 402260, China
关键词:
软脂酸钠内质网应激非酒精性脂肪性肝病凋亡
Keywords:
palmitate sodiumendoplasmic reticulum stressnon-alcoholic fatty liver diseaseapoptosis
分类号:
R575.202;R363;R329.25
文献标志码:
A
摘要:
目的     观察内质网应激(endoplasmic reticulum stress,ERS)蛋白在软脂酸钠诱导的脂肪变性L02肝细胞凋亡过程中的改变,探讨ERS与脂变肝细胞凋亡的关系。     方法     用软脂酸钠(饱和脂肪酸)诱导建立L02肝细胞脂肪变性模型。将细胞分为正常对照组和实验组(软脂酸钠72 μmol/L)。MTT法筛选软脂酸钠最佳作用浓度;油红O染色和甘油三酯(TG)试剂盒检测细胞内脂变程度;流式细胞术Annexin V-PE/7-AAD双染检测细胞凋亡率;Western blot检测GRP78、CHOP、磷酸化JNK(p-JNK)蛋白表达。     结果     软脂酸钠在体外能够诱导肝细胞发生脂肪变性以及脂变肝细胞凋亡,与对照组(1.38±0.42)比较,实验组48 h细胞凋亡率(5.95±0.37)显著增加(P<0.05)。Western blot检测示软脂酸钠刺激肝细胞12 h,GRP78表达(0.50±0.03)较0 h(0.37±0.02)明显增加,48 h(0.83±0.01)达高峰(P<0.05)。CHOP和p-JNK 表达与GRP78几乎同步增加。     结论     软脂酸钠对L02肝细胞具有较强的脂毒性,可以诱导肝细胞发生ERS反应。ERS上调CHOP和P-JNK表达可能在软脂酸钠诱导脂肪变性肝细胞凋亡中具有重要作用。
Abstract:
Objective     To observe the role of endoplasmic reticulum stress (ERS) proteins in palmitate sodium-induced apoptosis of human steatotic L02 hepatocytes and study the relation between ERS and apoptosis in steatotic hepatocytes.      Methods     A steatosis model of human L02 hepatocytes was induced by palmitate sodium. Hepatocytes were divided into normal control group and experimental group containing 72 μmol/L palmitate sodium. Optimal palmitate sodium concentration was selected by MTT assay. A triglyceride (TG) kit was used to detect lipid accumulation in hepatocytes with oil red O staining. Apoptosis of hepatocytes was detected by flow cytometry with Annexin V-PE/7-AAD staining. Expression of GRP78, CHOP and P-JNK was tdetected by Western blotting.      Results     Palmitate sodium induced steatosis and apoptosis of L02 hepatocytes in vitro. The apoptosis rate of L02 hepatocytes was significantly higher in experimental group than in control group 48 h after the model was induced by palmitate sodium(5.95±0.37 vs 1.38±0.42, P<0.05). Western blot analysis showed that the expression level of GRP78 was significantly higher at 12 h than at 0 h after the model was induced by palmitate sodium(0.50±0.03 vs 0.37±0.02, P<0.05)and reached its peak at 48 h(0.83±0.01). An almost similar increase was observed in expression of CHOP, P-JNK, and GRP78.      Conclusion     Palmitate sodium is rather lipotoxic to L02 hepatocytes and can induce ERS. ERS up-regulated CHOP and P-JNK expression may play an important role in palmitate sodium-induced apoptosis of steatotic L02 hepatocytes.

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更新日期/Last Update: 2011-09-26