|Table of Contents|

Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells

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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

Issue:
2018年第11期
Page:
954-958
Research Field:
基础医学
Publishing date:

Info

Title:

Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells

Author(s):

ZHOU Wei MA Li WAN Ying LI Fuxiang

Graduate School, Biomedical Analysis Center, Army Medical University (Third Military Medical University), Chongqing, 400038; Laboratory of Lipid & Glucose Metabolism, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016; Department of Intensive Care Medicine, General Hospital of Chengdu Military Command, Chengdu, Sichuan Province, 610083, China
 

Keywords:

nonalcoholic fatty liver disease AML12 cells Rab32 lipid metabolism

PACS:
Q591.5; R322.47; R329.26
DOI:
-
Abstract:

Objective    To determine the effect of regulation of Rab32 on the lipid metabolism in murine hepatic alpha mouse liver 12 (AML12) cells. Methods    Lentivirus expressing FCD-EYFP-Rab32 was transfected into AML12 cells for overexpression of Rab32, and CRISPR/Cas9 gene knockout technology was used to knockout Rab32 in the cells. Untreated AML12 cells were used as normal control. The protein level of Rab32 was determined by Western blotting. The intracellular lipid droplets were observed by confocal microscopy after Nile red fluorescent staining. The contents of intracellular triglyceride (TG) and total cholesterol (TC) were measured by corresponding quantitation kits. Cell apoptosis was assayed by flow cytometry. Results    Compared with the normal control cells, the protein level of Rab32 was significantly increased in the overexpressed cells but decreased in the knockout cells (P<0.01). The number (21.91±14.32 vs 33.89±17.31, P<0.01) and size (16.82±14.37 vs 22.27±14.10 μm2, P<0.01) of lipid droplets per cell were significantly decreased in the overexpressed cells when compared with the untreated cells, while the intracellular contents of TG (104.03±12.28 vs 130.94±4.21 nmol, P<0.01) and TC (46.92±1.26 vs 81.11±0.65 μg, P<0.01) were decreased. Whereas, the knockout cells had larger lipid droplet number (58.23±42.28, P<0.01) and total area (53.31±36.33 μm2, P<0.01), and higher intracellular TG (159.03±8.85 nmol, P<0.01) and TC (93.38±3.33 μg, P<0.01). No obvious difference was found in cell apoptosis among the 3 groups of cells (P>0.05). Conclusion    Rab32 promotes the lipid metabolism in the AML12 cells, and the molecule may be regarded as a new target in treatment of nonalcoholic fatty

References:

[1]FARRELL G C,WONG V W,CHITTURI S. NAFLD in Asia—as common and important as in the West[J]. Nat Rev Gastroenterol Hepatol,2013,10(5):307-318. DOI:10.1038/nrgastro.2013.34.
[2]YAN J,XIE W,OU W N,et al. Epidemiological survey and risk factor analysis of fatty liver disease of adult residents,Beijing,China[J]. J Gastroenterol Hepatol,2013,28(10):1654-1659. DOI:10.1111/jgh.12290.
[3]BUZZETTI E,PINZANI M,TSOCHATZIS E A. The multiplehit pathogenesis of non-alcoholic fatty liver disease (NAFLD)[J]. Metabolism,2016,65(8):1038-1048. DOI:10.1016/j.metabol.2015.12.012.
[4]PRASHAR A,SCHNETTGER L,BERNARD E M,et al.RabGTPases in immunity and inflammation[J]. Front Cell Infect Microbiol, 2017, 7:435. DOI:10.3389/fcimb.2017.00435.
[5]SCHROEDER B,SCHULZE R J,WELLER S G,et al.The small GTPase Rab7 as a central regulator of hepatocellular lipophagy[J]. Hepatology,2015,61(6):1896-1907. DOI:10.1002/hep.27667.
[6]LI Z,SCHULZE R J,WELLER S G,et al.A novel Rab10EHBP-EHD2 complex essential for the autophagicengulfment of lipid droplets[J]. Sci Adv,2016,2(12):e1601470. DOI:10.1126/sciadv.1601470.
[7]LI C,LUO X,ZHAO S,et al.COPITRAPPII activates Rab18 and regulates its lipid droplet association[J]. EMBO J, 2017,36(4):441-457. DOI:10.15252/embj.201694866.
[8]COHENSOLAL K A,SOOD R,MARIN Y,et al.Identification and characterization of mouse Rab32 by mRNA and protein expression analysis[J]. Biochim Biophys Acta,2003,1651(1/2):68-75.
[9]HAILE Y,DENG X,ORTIZSANDOVAL C,et al. Rab32 connects ER stress to mitochondrial defects in multiple sclerosis[J]. J Neuroinflammation,2017,14(1): 19. DOI:10.1186/s129740160788z.
[10]KRAHMER N,HILGER M,KORY N,et al.Protein correlation profiles identify lipid droplet proteins with high confidence[J]. Mol Cell Proteomics,2013,12(5):1115-1126. DOI:10.1074/mcp.M112.020230.
[11]LARSSON S,RESJO S,GOMEZ M F,et al.Characterization of the lipid droplet proteome of a clonal insulinproducing betacell line(INS1 832/13)[J]. J Proteome Res, 2012, 11(2):1264-1273. DOI:10.1021/pr200957p.
[12]RASINENI K,MCVICKER B L,TUMA D J,et al.RabGTPases associate with isolated lipid droplets(LDs) and show altered content after ethanol administration:potential role in alcoholimpaired LD metabolism[J]. Alcohol Clin Exp Res,2014,38(2):327-335. DOI:10.1111/acer.12271.
[13]BUI M,GILADY S Y,FITZSIMMONS R E,et al. Rab32 modulates apoptosis onset and mitochondriaassociated membrane (MAM) properties[J]. J Biol Chem,2010,285(41):31590-31602. DOI:10.1074/jbc.M110.101584.
[14]BOREN J,BRINDLE K M. Apoptosisinduced mitochondrial dysfunction causes cytoplasmic lipid droplet formation[J]. Cell Death Differ,2012,19(9):1561-1570. DOI:10.1038/cdd.2012.34.
[15]WANG C,LIU Z,HUANG X. Rab32 is important for autophagy and lipid storage in drosophila[J]. PLoS ONE, 2012, 7(2):e32086. DOI:10.1371/journal.pone.0032086.
[16]LI Q,WANG J,WAN Y,et al. Depletion of Rab32 decreases intracellular lipid accumulation and induces lipolysis through enhancing ATGL expression in hepatocytes[J]. Biochem Biophy Res Commun,2016,471(4):492-496. DOI:10.1016/j.bbrc.2016.02.047.
[17]ONAL G,KUTLU O,GOZUACIK D,et al.Lipiddroplets in health and disease[J]. Lipids Health Dis,2017,16(1): 128. DOI:10.1186/s1294401705217.

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Last Update: 2018-06-13