[1]房殿亮,宁波,沈薇,等.干扰SREBP-1c对内质网应激状态下L02和HepG2肝细胞脂质沉积的影响[J].陆军军医大学学报(原第三军医大学学报),2013,35(06):513-517.
 Fang Dianliang,Ning Bo,Shen Wei,et al.SREBP-1c gene interference decreases lipid accumulation in L02 and HepG2 hepatocytes under endoplasmic reticulum stress[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(06):513-517.
点击复制

干扰SREBP-1c对内质网应激状态下L02和HepG2肝细胞脂质沉积的影响(/HTML )
分享到:

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:
35卷
期数:
2013年第06期
页码:
513-517
栏目:
论著
出版日期:
2013-03-30

文章信息/Info

Title:
SREBP-1c gene interference decreases lipid accumulation in L02 and HepG2 hepatocytes under endoplasmic reticulum stress
作者:
房殿亮宁波沈薇万颖
重庆医科大学附属第二医院消化内科
Author(s):
Fang Dianliang Ning Bo Shen Wei Wan Ying
Department of Gastroenterology, Second Affiliated Hospital, Chongqing Medical University, Chongqing, 400010, China
关键词:
非酒精性脂肪肝SREBP-1cFAS内质网应激RNA干扰
Keywords:
non-alcoholic fatty liver disease SREBP-1cfatty acid synthase endoplasmic reticulum stress RNA interference
分类号:
R322.47;R349.15;R394.3
文献标志码:
A
摘要:
目的      构建SREBP-1c基因的miRNA真核表达载体,观察其对内质网应激状态下L02和HepG2肝细胞脂质沉积的影响。      方法      设计并构建SREBP-1c基因的3个miRNA干扰载体,经测序鉴定miRNA载体构建成功后转染肝细胞,荧光显微镜下观察绿色荧光蛋白表达。Western blot检测转染后SREBP-1c的表达;甘油三酯测定和油红O染色检测细胞内脂质沉积;Western blot检测SREBP-1c下游脂代谢相关基因脂肪酸合成酶(fatty acid synthase,FAS)和乙酰辅酶A羧化酶(acetyl coA  carboxylase,ACC1)的表达。      结果      靶向干扰SREBP-1c的miRNA真核表达载体构建成功。Western blot结果显示,转染SREBP-1c-1miRNA和SREBP-1c-3miRNA载体后,L02和HepG2细胞内SREBP-1c蛋白水平表达均明显减低(P<0.05),SREBP-1c-1miRNA的干扰效果最好(P<0.05)。与对照组相比,转染SREBP-1c-1miRNA载体后L02和HepG2肝细胞内甘油三酯含量均明显降低(P<0.05),细胞内脂滴明显减少;FAS和ACC1蛋白的表达明显降低(P<0.05)。      结论      SREBP-1c基因沉默通过FAS/ACC1降低了内质网应激状态下肝细胞脂质合成。
Abstract:
Objective      To construct miRNA eukaryotic expression vectors targeting SREBP-1c and determine the effect of SREBP-1c gene knockdown on lipid accumulation in L02 and HepG2 hepatocytes under endoplasmic reticulum stress.       Methods      Three miRNA targeting SREBP-1c gene, named SREBP-1c-1-miRNA, SREBP-1c-2-miRNA and SREBP-1c-3-miRNA, and a non-homologous negative control expression vector (HK-miRNA) were designed and constructed which were identified by DNA sequencing analysis. After 3 miRNA expression vectors were transiently transfected into hepatocytes, fluorescence microscopy was applied to observe EGFP to confirm the transfection. Western blotting was used to determine expression of SREBP-1c. Lipid deposition in each group was observed by triglyceride content detection and oil red O staining. Western blotting was also applied to measure the protein levels of fatty acid synthase (FAS) and acetyl CoA carboxylase (ACC1).       Results      Four miRNA expression vectors were constructed successfully. After 24 hours’ transfection, EGFP expression was visible under fluorescence microscope. The protein level of SREBP-1c was decreased significantly after transfected with SREBP-1c-1miRNA and SREBP-1c-3miRNA, when compared with control group (P<0.05). SREBP-1c-1miRNA was identified with most powerful interference (P<0.05). Compared with the control group, TG content (P<0.05) and lipid droplet were significantly decreased, and FAS and ACC1 proteins were also inhibited in SREBP-1c inference group (P<0.05).       Conclusion      SREBP-1c gene interference suppresses lipid synthesis through down-regulating FAS and ACC1 in L02 and HepG2 hepatocytes under endoplasmic reticulum stress.

相似文献/References:

[1]高燕翔,张勇,刘裕,等.SREBP-1c在白藜芦醇预防大鼠非酒精性脂肪肝发生中的作用[J].陆军军医大学学报(原第三军医大学学报),2015,37(17):1704.
 Gao Yanxiang,Zhang Yong,Liu Yu,et al.SREBP-1c is involved in preventive effect of resveratrol on pathogenesis of nonalcoholic fatty liver disease in rats[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(06):1704.
[2]杨朝霞,代东伶,沈薇.虫草菌丝和还原型谷胱甘肽对非酒精性脂肪肝大鼠模型治疗效果的研究[J].陆军军医大学学报(原第三军医大学学报),2007,29(22):2176.
 YANG Zhao-xia,DAI Dong-ling,SHEN Wei.Effect of cordyceps sinensis and reduced glutathione on experimental mouse model with non-alcoholic fatty liver disease[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(06):2176.
[3]史洪涛,李陶,陈东风.肝细胞色素氧化酶Ⅰ在非酒精性脂肪肝大鼠形成中的作用[J].陆军军医大学学报(原第三军医大学学报),2006,28(02):160.
[4]张贵明,倪向敏,崔涵强,等.雌马酚干预对高脂饮食诱导的去卵巢大鼠非酒精性脂肪肝的影响[J].陆军军医大学学报(原第三军医大学学报),2022,44(21):2129.
 ZHANG Guiming,NI Xiangmin,CUI Hanqiang,et al.Effect of equol intervention on high-fat diet-induced non-alcoholic fatty liver disease in ovariectomized rats[J].J Amry Med Univ (J Third Mil Med Univ),2022,44(06):2129.
[5]王万东,陈东风.非酒精性脂肪变性肝细胞模型中ATF4基因的表达及其意义[J].陆军军医大学学报(原第三军医大学学报),2010,32(14):1487.
 Wang Wandong,Chen Dongfeng.Expression and significance of activating transcription factor 4 in oleic acid-induced hepatocyte steatosis in vitro[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(06):1487.
[6]周婷婷,秦波,郑天鹏,等.非酒精性脂肪肝患者血浆FGF21水平及与肥胖、胰岛素抵抗关系研究[J].陆军军医大学学报(原第三军医大学学报),2010,32(03):265.
 Zhou Tingting,Qin Bo,Zheng Tianpeng,et al.Correlation between plasma fibroblast growth factor 21 level, obesity and insulin resistance in nonalcoholic fatty liver disease patients[J].J Amry Med Univ (J Third Mil Med Univ),2010,32(06):265.
[7]曾志华,曾明慧,冯雯琪,等.电针阻断胰岛素抵抗及过氧化反应治疗非酒精性脂肪肝的实验研究[J].陆军军医大学学报(原第三军医大学学报),2008,30(11):1055.
 ZENG Zhi-hua,ZENG Ming-hui,FENG Wen-qi,et al.Electroacupuncture treats rat non-alcoholic fatty liver by blocking insulin resistance and peroxidation[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(06):1055.
[8]田晓媛,张乾勇,冉莉,等.不同膳食脂肪酸对肝细胞SREBP-1c表达及脂代谢的影响[J].陆军军医大学学报(原第三军医大学学报),2012,34(01):66.
 Tian Xiaoyuan,Zhang Qianyong,Ran Li,et al.Effect of different dietary fatty acids on expression of sterol-regulatory elementary binding protein-1c and lipid metabolism in hepatic cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(06):66.
[9]邱烈旺,顾陆昀,吕琳,等.水甘油通道蛋白在非酒精性脂肪变性肝细胞模型中的表达及意义[J].陆军军医大学学报(原第三军医大学学报),2012,34(07):622.
 Qiu Liewang,Gu Luyun,Lu Lin,et al.Expression of aquaglyceroporins in nonalcoholic hepatocyte steatosis and its significance[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(06):622.
[10]肖凯,朱晟易,童钰铃,等.普洱茶水提取物对大鼠非酒精性脂肪性肝病的干预作用[J].陆军军医大学学报(原第三军医大学学报),2013,35(18):1992.

更新日期/Last Update: 2013-03-19