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物理方法构建肝细胞自噬模型

朱学敏 孟庆华

引用本文:
Citation:

物理方法构建肝细胞自噬模型

DOI: 10.3969/j.issn.1001-5256.2016.08.027
基金项目: 

国家自然科学基金资助项目(81470877); 

详细信息
  • 中图分类号: R575

Hepatocyte autophagy model established by physical method

Research funding: 

 

  • 摘要:

    目的应用物理方法构建正常人肝细胞株7702细胞的低氧饥饿自噬模型,为研究细胞自噬对肝脏功能的影响机制奠定基础。方法选用7702细胞,使用完全DMEM培养基,在37℃和5%CO2培养箱中培养(正常对照组),使用Binder三气培养箱,设置温度为37℃,CO2浓度为5%、O2浓度为0.3%来提供缺氧环境,使用无血清DMEM培养细胞来诱导饥饿,将其分为低氧饥饿6、12、18和24 h 4个时间组。用Western Blot检测正常对照组和不同时间点实验组的Beclin-1、Atg5以及LC3蛋白表达水平的变化;通过实时荧光定量PCR法检测各组Beclin-1 mRNA、Atg5 mRNA的表达;转染LC3质粒后利用免疫荧光法观察各组细胞发生自噬的情况。计量资料多组间比较采用方差分析,进一步两两比较采用LSD-t检验;计数资料组间比较采用χ2检验。结果低氧饥饿处理6 h组的Beclin-1、Atg5及LC3的蛋白表达量高于正常对照组及其他处理组;低氧饥饿6 h组较其他各组的Beclin-1mRNA和Atg5 mRNA的表达量显著增高,且其增高最显著,差异均有统计学意义(P值均<0.05)...

     

  • [1]LIU YP,DONG L.Research advances in the role of autophagy in the development,progression,and treatment of liver cancer[J].Int J Dig Dis,2011,31(6):351-353.(in Chinese)刘亚萍,董蕾.自噬与肝癌发生发展和治疗的研究进展[J].国际消化病杂志,2011,31(6):351-353.
    [2]HUANG LW,XU LM.Relationship between liver diseases and autophagy[J].J Clin Hepatol,2014,30(2):186-188.(in Chinese)黄兰蔚,徐列明.细胞自噬在肝脏疾病中的作用[J].临床肝胆病杂志,2014,30(2):186-188.
    [3]GEBHARDT R,COFFER PJ.Hepatic autophagy is differentially regulated in periportal and pericentral zones-a general mechanism relevant for other tissues?[J].Cell Commun Signal,2013,11(1):21.
    [4]CHRISTIAN P,SACCO J,ADELI K.Autophagy:emerging roles in lipid homeostasis and metabolic control[J].Biochim Biophys Acta,2013,1831(4):819-824.
    [5]MA B,CAO W,LI W,et al.Dapper1 promotes autophagy by enhancing the Beclin1-Vps34-Atg14L complex formation[J].Cell Res,2014,24(8):912-924.
    [6]ZHANG DX,ZHANG JP,HU JY,et al.The potential regulatory roles of NAD+and its metabolism in autophagy[J].Metabolism,2016,65(4):456-462.
    [7]STEELE S,BRUNTON J,KAWULA T.The role of autophagy in intracellular pathogen nutrient acquisition[J].Front Cell Infect Microbiol,2015,5:51.
    [8]ROLANDO M,ESCOLL P,NORA T,et al.Legionella pneumophila S1P-lyase targets host sphingolipid metabolism and restrains autophagy[J].Proc Natl Acad Sci U S A,2016,113(7):1901-1906.
    [9]VAKIFAHMETOGLU-NORBERG H,XIA HG,YUAN J.Pharmacologic agents targeting autophagy[J].J Clin Invest,2015,125(1):5-13.
    [10]TANG Q,BU WH,WANG DD,et al.Advance research on interaction between autophagy and apoptosis and its influence in doevelopment of tumors[J].J Jilin Univ:Med Edit,2015,41(6):1303-1306.(in Chinese)唐琪,布文奂,王丹丹,等.自噬与凋亡的相互作用及其对肿瘤发展过程影响的研究进展[J].吉林大学学报:医学版,2015,41(6):1303-1306.
    [11]HARDAS SS,SULTANA R,WARRIER G,et al.Rat hippocampal responses up to 90 days after a single nanoceria dose extends a hierarchical oxidative stress model for nanoparticle toxicity[J].Nanotoxicology,2014,8(Suppl 1):155-166.
    [12]SONG W,SOO LEE S,SAVINI M,et al.Ceria nanoparticles stabilized by organic surface coatings activate the lysosome-autophagy system and enhance autophagic clearance[J].ACS Nano,2014,8(10):10328-10342.
    [13]HUANG J,BRUMELL JH.Bacteria-autophagy interplay:a battle for survival[J].Nat Rev Microbiol,2014,12(2):101-114.
    [14]ESCOLL P,ROLANDO M,BUCHRIESER C.Modulation of host autophagy during bacterial infection:sabotaging host munitions for pathogen nutrition[J].Front Immunol,2016,7:81.
    [15]FU ZY.Molecular mechanism and regulating factors of autophagy[J].J Clin Exp Med,2015,14(17):1488-1489.(in Chinese)付智勇.自噬的分子机制及其调控因素[J].临床和实验医学杂志,2015,14(17):1488-1489.
    [16]NOVAK I,KIRKIN V,McE WAN DG,et al.Nix is a selective autophagy receptor for mitochondrial clearance[J].EMBO Rep,2010,11(1):45-51.
    [17]RUBINSZTEIN DC,MARIO G,KROEMER G.Autophagy and aging[J].Cell,2011,146(5):682-695.
    [18]LIN CF,KUO YT,CHEN TY,et al.Quercetin-rich guava(psidium guajava)juice in combination with trehalose reduces autophagy,apoptosis and pyroptosis formation in the kidney and pancreas of type II diabetic rats[J].Molecules,2016,21(3):e334.
    [19]FRANOIS A,JULIAN A,RAGOT S,et al.Correction:inflammatory stress on autophagy in peripheral blood mononuclear cells from patients with alzheimer's disease during 24 months of followup[J].PLoS One,2016,11(3):e0143933.
    [20]CHUN SK,GO K,YANG MJ,et al.Autophagy in ischemic livers:a critical role of sirtuin 1/mitofusin 2 axis in autophagy induction[J].Toxicol Res,2016,32(1):35-46.
    [21]PROSKE J,WALTER L,BUMES E,et al.Adaptive immune response to and survival effect of temozolomide-and valproic acidinduced autophagy in glioblastoma[J].Anticancer Res,2016,36(3):899-905.
    [22]WANG KF,ZHU YK,MENG QH.Research advances in oxidative stress and mitochondrial permeability transition in liver failure[J].JClin Hepatol,2013,29(9):711-714.(in Chinese)王克菲,朱跃科,孟庆华.氧化应激、线粒体通透性转换与肝衰竭的相关性[J].临床肝胆病杂志,2013,2(9):711-714.
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  • 收稿日期:  2016-02-19
  • 出版日期:  2016-08-20
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