
浏览全部资源
扫码关注微信
收稿:2018-02-16,
纸质出版:2018-06
移动端阅览
周围, 马莉, 万瑛, 等. Rab32对小鼠肝细胞系AML12细胞内脂质代谢的影响[J]. 第三军医大学学报, 2018,40(11):954-958.
Wei ZHOU, Li MA, Ying WAN, et al. Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells[J]. Journal of Third Military Medical University, 2018, 40(11): 954-958.
周围, 马莉, 万瑛, 等. Rab32对小鼠肝细胞系AML12细胞内脂质代谢的影响[J]. 第三军医大学学报, 2018,40(11):954-958. DOI: 10.16016/j.1000-5404.201802048.
Wei ZHOU, Li MA, Ying WAN, et al. Effect of Rab32 on lipid metabolism in mouse hepatic AML12 cells[J]. Journal of Third Military Medical University, 2018, 40(11): 954-958. DOI: 10.16016/j.1000-5404.201802048.
Objective
2
To determine the effect of regulation of Rab32 on the lipid metabolism in murine hepatic alpha mouse liver 12 (AML12) cells.
Methods
2
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
2
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 μm
2
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 μm
2
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
2
Rab32 promotes the lipid metabolism in the AML12 cells
and the molecule may be regarded as a new target in treatment of nonalcoholic fatty liver disease.
FARRELL G C WONG V W CHITTURI S NAFLD in Asia-as common and important as in the West Nat Rev Gastroenterol Hepatol 2013 10 5 307 318 10.1038/nrgastro.2013.34 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.
YAN J XIE W OU W N Epidemiological survey and risk factor analysis of fatty liver disease of adult residents, Beijing, China J Gastroenterol Hepatol 2013 28 10 1654 1659 http://cn.bing.com/academic/profile?id=bcd81ada9e4030a351e2254a75262bcc&encoded=0&v=paper_preview&mkt=zh-cn 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.
BUZZETTI E PINZANI M TSOCHATZIS E A The multiple-hit pathogenesis of non-alcoholic fatty liver disease (NAFLD) Metabolism 2016 65 8 1038 1048 10.1016/j.metabol.2015.12.012 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.
PRASHAR A SCHNETTGER L BERNARD E M RabGTPases in immunity and inflammation Front Cell Infect Microbiol 2017 7 435 10.3389/fcimb.2017.00435 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.
SCHROEDER B SCHULZE R J WELLER S G The small GTPase Rab7 as a central regulator of hepatocellular lipophagy Hepatology 2015 61 6 1896 1907 10.1002/hep.27667 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.
LI Z SCHULZE R J WELLER S G A novel Rab10-EHBP1-EHD2 complex essential for the autophagicengulfment of lipid droplets Sci Adv 2016 2 12 e1601470 10.1126/sciadv.1601470 LI Z, SCHULZE R J, WELLER S G, et al .A novel Rab10-EHBP1-EHD2 complex essential for the autophagicengulfment of lipid droplets[J ] . Sci Adv, 2016, 2(12):e1601470. DOI:10.1126/sciadv.1601470.
LI C LUO X ZHAO S COPI-TRAPPII activates Rab18 and regulates its lipid droplet association EMBO J 2017 36 4 441 457 10.15252/embj.201694866 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.
COHEN-SOLAL K A SOOD R MARIN Y Identification and characterization of mouse Rab32 by mRNA and protein expression analysis Biochim Biophys Acta 2003 1651 1/2 68 75 http://cn.bing.com/academic/profile?id=5816a98250bcf5b3b090ada9d0aa281d&encoded=0&v=paper_preview&mkt=zh-cn 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.
HAILE Y DENG X ORTIZ-SANDOVAL C Rab32 connects ER stress to mitochondrial defects in multiple sclerosis J Neuroinflammation 2017 14 1 19 10.1186/s12974-016-0788-z 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.
KRAHMER N HILGER M KORY N Protein correlation profiles identify lipid droplet proteins with high confidence Mol Cell Proteomics 2013 12 5 1115 1126 10.1074/mcp.M112.020230 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.
LARSSON S RESJO S GOMEZ M F Characterization of the lipid droplet proteome of a clonal insulin-producing beta-cell line(INS-1 832/13) J Proteome Res 2012 11 2 1264 1273 10.1021/pr200957p 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.
RASINENI K MCVICKER B L TUMA D J RabGTPases associate with isolated lipid droplets(LDs) and show altered content after ethanol administration:potential role in alcohol-impaired LD metabolism Alcohol Clin Exp Res 2014 38 2 327 335 10.1111/acer.12271 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.
BUI M GILADY S Y FITZSIMMONS R E Rab32 modulates apoptosis onset and mitochondria-associated membrane (MAM) properties J Biol Chem 2010 285 41 31590 31602 10.1074/jbc.M110.101584 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.
BOREN J BRINDLE K M Apoptosis-induced mitochondrial dysfunction causes cytoplasmic lipid droplet formation Cell Death Differ 2012 19 9 1561 1570 10.1038/cdd.2012.34 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.
WANG C LIU Z HUANG X Rab32 is important for autophagy and lipid storage in drosophila PLoS ONE 2012 7 2 e32086 10.1371/journal.pone.0032086 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.
LI Q WANG J WAN Y Depletion of Rab32 decreases intracellular lipid accumulation and induces lipolysis through enhancing ATGL expression in hepatocytes Biochem Biophy Res Commun 2016 471 4 492 496 10.1016/j.bbrc.2016.02.047 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.
ONAL G KUTLU O GOZUACIK D Lipiddroplets in health and disease Lipids Health Dis 2017 16 1 128 10.1186/s12944-017-0521-7 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.
0
浏览量
0
下载量
0
CSCD
关联资源
相关文章
相关作者
相关机构
京公网安备11010802024621