
浏览全部资源
扫码关注微信
1.400037 重庆,第三军医大学新桥医院:消化内科
2.400037 重庆,第三军医大学新桥医院:内分泌科
[郑宏庭,E-mail:fnf7703@hotmail.comZHENG Hongting, E-mail:fnf7703@hotmail.com
[杨仕明,E-mail:shimingyang@yahoo.comYANG Shiming, E-mail:shimingyang@yahoo.com
收稿:2017-07-17,
修回:2017-08-14,
纸质出版:2017-11-15
移动端阅览
宋铃榆, 廖晓玉, 郑宏庭, 等. 溶酶体相关膜蛋白3过表达促进肝细胞脂质沉积[J]. 第三军医大学学报, 2017,39(21):2059-2065.
Lingyu SONG, Xiaoyu LIAO, Hongting ZHENG, et al. Overexpression of LAMP3 promotes lipid metabolism in liver cells[J]. Journal of Third Military Medical University, 2017, 39(21): 2059-2065.
宋铃榆, 廖晓玉, 郑宏庭, 等. 溶酶体相关膜蛋白3过表达促进肝细胞脂质沉积[J]. 第三军医大学学报, 2017,39(21):2059-2065. DOI: 10.16016/j.1000-5404.201707111.
Lingyu SONG, Xiaoyu LIAO, Hongting ZHENG, et al. Overexpression of LAMP3 promotes lipid metabolism in liver cells[J]. Journal of Third Military Medical University, 2017, 39(21): 2059-2065. DOI: 10.16016/j.1000-5404.201707111.
目的
2
探讨溶酶体相关膜蛋白3 (lysosome associated membrane protein 3
LAMP3)对肝细胞脂质代谢的作用。
方法
2
以肝癌细胞系Huh7为研究对象,利用qRT-PCR及Western blot法检测游离脂肪酸(free fatty acids,FFAs)0.4 mmol/L处理后肝细胞LAMP3表达变化;过表达LAMP3后,分别使用油红染色及甘油三酯检测试剂盒检测肝细胞内脂滴及甘油三酯沉积情况;采用ATP及β-羟基丁酸检测试剂盒检测细胞内ATP及β-羟基丁酸含量变化;过表达LAMP3,使用qRT-PCR、Western blot法检测脂质合成相关基因PPARγ与SCD-1及脂质分解相关基因CPT1A与PGC-1α表达变化。
结果
2
FFAs处理后肝细胞Huh7中LAMP3表达显著上调(
P
<
0.05);过表达LAMP3可显著促进肝细胞内脂滴和甘油三酯的沉积(
P
<
0.05);过表达LAMP3对肝细胞内ATP及β-羟基丁酸含量的影响差异无统计学意义(
P
>
0.05);过表达LAMP3可促进PPARγ及SCD-1表达(
P
<
0.05),但对CPT1A、PGC-1α表达的影响差异无统计学意义(
P
>
0.05)。
结论
2
LAMP3可通过调控脂肪合成促进肝细胞内脂质沉积。
Objective
2
To illustrate the role of lysosome associated membrane protein 3 (LAMP3) in hepatocellular lipid metabolism.
Methods
2
After free fatty acids (FFAs
0.4 mmol/L) were used to treat human hepatocellular carcinoma cell line Huh7
the expression of LAMP3 was detected by qRT-PCR and Western blotting. Then
oil red staining was adopted to observe the lipid droplets in the FFAs-treated Huh7 cells
and intracellular triglyceride level was detected at the same condition. ATP content and β-hydroxybutyrate concentration were measured by the corresponding kits respectively. Finally
the expression of lipogenic molecules
PPARγ and SCD-1
and those involved in lipolysis
CPT1A and PGC-1α
at mRNA and protein levels were detected in Huh7 cells after FFAs treatment by qRT-PCR and Western blotting
respectively.
Results
2
FFAs treatment resulted in the up-regulation of LAMP3 in Huh7 cells (
P
<
0.05). The treatment also promoted the deposition of intracellular lipid droplets and triglycerides in the cells (
P
<
0.05). However
it had no effect on the intracellular ATP and β-hydroxybutyrate levels (
P
>
0.05). FFAs-treatment induced up-regulation of LAMP3 also enhanced the expression of PPARγ and SCD-1 (
P
<
0.05)
though showing no effect on the levels of CPT1A and PGC-1α (
P
>
0.05).
Conclusion
2
LAMP3 promotes deposition of lipid droplets
via
regulating fat synthesis in liver cells.
WILLAMS C D, STENGEL J, ASIKE M I, et al. Prevalence of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis among a largely middle-aged population utilizing ultrasound and liver biopsy: a prospective study[J]. Gastroenterology, 2011, 140(1):124-131.DOI:10.1053/j.gastro.2010.09.038.
SCHREUDER T C, VERWER B J, VAN NIEUWKERK C M, et al. Nonalcoholic fatty liver disease: an overview of current insights in pathogenesis, diagnosis and treatment. World journal of gastroenterology[J]. Word J Gastroenterol, 2008, 14(16):2474-2486.DOI:10.3748/wjg.14.2474.
FUCHS C D, CLAUDEL T, KUMARI P, et al. Absence of adipose triglyceride lipase protects from hepatic endoplasmic reticulum stress in mice[J]. Hepatology, 2012, 56(1):270-280.DOI:10.1002/hep.25601.
EKSTEDT M, HAGSTROM H, NASR P, et al. Fibrosis stage is the strongest predictor for disease-specific mortality in NAFLD after up to 33 years of follow-up[J]. Hepatology, 2015, 61(5):1547-1554.DOI:10.1002/hep.27368.
Fu T, Zhao X, Evans R M. Liver cancer checks in when bile acid clocks out[J]. Cancer Cell, 2016, 30(6):827-828.DOI:10.1016/j.ccell.2016.11.012.
WANG S, KAUFMAN R J. How does protein misfolding in the endoplasmic reticulum affect lipid metabolism in the liver[J]. Curr Opin Lipidol, 2014, 25(2):125-132.DOI:10.1097/mol.0000000000000056.
MA C, KESARWALA A H, EGGERT T, et al. NAFLD causes selective CD4(+) T lymphocyte loss and promotes hepatocarcinogenesis[J]. Nature, 2016, 531(7593):253-257.DOI:10.1038/nature16969.
SINGH R, KAUSHIK S, WANG Y, et al. Autophagy regulates lipid metabolism[J].Nature, 2009, 458(7242):1131-1135.DOI:10.1038/nature07976.
SINHA RA, YOU SH, ZHOU J, et al. Thyroid hormone stimulates hepatic lipid catabolism via activation of autophagy[J ] . J Clin Investi, 2012, 122(7):2428-2438.DOI:10.1172/JCI60580.
JU X, YAN Y, LIU Q, et al. Neuraminidase of influenza A virus binds lysosome-associated membrane proteins directly and induces lysosome rupture[J]. J Virol, 2015, 89(20):10347-10358.DOI:10.1128/jvi.01411-15.
YASUDA-YAMAHARA M, KUME S, YAMAHARA K, et al. Lamp-2 deficiency prevents high-fat diet-induced obese diabetes via enhancing energy expenditure[J]. Biochem Biophys Res Commun, 2015, 465(2):249-255.DOI:10.1016/j.bbrc.2015.08.010.
TSENG Y H, KE P Y, LIAO C J, et al. Chromosome 19 open reading frame 80 is upregulated by thyroid hormone and modulates autophagy and lipid metabolism[J]. Autophagy, 2014, 10(1):20-31.DOI:10.4161/auto.26126.
WEICHERT N, KALTENBORN E, HECTOR A, et al. Some ABCA3 mutations elevate ER stress and initiate apoptosis of lung epithelial cells[J]. Respiratory research, 2011, 12(1):4.DOI:10.1186/1465-9921-12-4.
LIAO X, CHEN Y, LIU D, et al. High expression of LAMP3 is a novel biomarker of poor prognosis in patients with esophageal squamous cell carcinoma[J]. Int J Mol Sci, 2015, 16(8):17655-17667.DOI:10.3390/ijms160817655.
KANAO H, ENOMOTO T, KIMURA T, et al. Overexpression of LAMP3/TSC403/DC-LAMP promotes metastasis in uterine cervical cancer[J]. Cancer Res, 2005, 65(19):8640-9645.DOI:10.1158/0008-5472.CAN-04-4112.
MUJCIC H, NAGELKERKE A, ROUSCHOP K M, et al. Hypoxic activation of the PERK/eIF2alpha arm of the unfolded protein response promotes metastasis through induction of LAMP3[J]. Clinical Cancer Research, 2013, 19(22):6126-6137.DOI:10.1158/1078-0432.CCR-13-0526.
WANG C, CHI Y, LI J, et al. FAM3A activates PI3K p110alpha/Akt signaling to ameliorate hepatic gluconeogenesis and lipogenesis[J]. Hepatology, 2014, 59(5):1779-1790.DOI:10.1002/hep.26945.
ARSENIJEVIC D, CAJOT J F, FELLAY B, et al. Uninephrectomy-induced lipolysis and low-grade inflammation are mimicked by unilateral renal denervation[J]. Front Physiol, 2016, 7:227.DOI:10.3389/fphys.2016.00227.
LU J, MA H, LIAN S, et al. Clinical significance and prognostic value of the expression of LAMP3 in oral squamous cellcarcinoma[J]. Dis Markers, 2017, 2017:1218254.DOI:10.1155/2017/1218254.
WANG D, CAO X, ZHANG Y, et al. LAMP3 expression correlated with poor clinical outcome in human ovarian cancer[J]. Tumour Biol, 2017, 39(3):10104283176 95014.DOI:10.1177/1010428317695014.
AHMADIAN M, SUHJ M, HAH N, et al. PPARγ signaling and metabolism: the good, the bad and the future[J]. Nat Med, 2013, 19(5):557-566.DOI:10.1038/nm.3159.
KANURI G, LANDMANN M, PRIEBS J, et al. Moderate alcohol consumption diminishes the development of non-alcoholic fatty liver disease (NAFLD) in ob/ob mice[J]. Eur J Nutr, 2016, 55(3):1153-1164.DOI:10.1007/s00394-015-0929-7.
PETTINELLI P, VIDELA L A. Up-regulation of PPAR-γ mRNA expression in the liver of obese patients: an additional reinforcing lipogenic mechanism to SREBP-1c induction[J]. J Clin Endocrinol Metab, 2011, 96(5):1424-1430.DOI:10.1210/jc.2010-2129.
MATSUSUE K, HALUZIK M, LAMBERT G, et al. Liver-specific disruption of PPARγ in leptin-deficient mice improves fatty liver but aggravates diabetic phenotypes[J]. J Clin Invest, 2003, 111(5):737-747.DOI:10.1172/JCI200317223.
COHEN P, MIYAZAKI M, SOCCI N D, et al. Role for stearoyl-CoA desaturase-1 in leptin-mediated weight loss[J]. Science, 2002, 297(5579):240-243.DOI:10.1126/science.1071527.
GONZALEZ-RODRIGUEZ A, MAYORAL R, AGRA N, et al. Impaired autophagic flux is associated with increased endoplasmic reticulum stress during the development of NAFLD[J]. Cell Dath Dsease, 2014, 5(4):e1179.DOI: 10.1038/cddis.2014.162.
ZHANG X Q, XU C F, YU C H, et al. Role of endoplasmic reticulum stress in the pathogenesis of nonalcoholic fatty liver disease[J]. World J Gastroenterol, 2014, 20(7):1768-1776.DOI:10.3748/wjg.v20.i7.1768.
OZCAN U, CAO Q, YILMAZ E, et al. Endoplasmic reticulum stress links obesity, insulin action, and type 2 diabetes[J]. Science, 2004, 306(5695):457-461.DOI:10.1126/science.1103160.
GREGOR M F, YANG L, FABBRINI E, et al. Endoplasmic reticulum stress is reduced in tissues of obese subjects after weight loss[J]. Diabetes, 2009, 58(3):693-700.DOI:10.2337/db08-1220.
BOBROVNIKOVA-MARJON E, HATZIVASSILIOU G, GRIGORIADOU C, et al. PERK-dependent regulation of lipogenesis during mouse mammary gland development and adipocyte differentiation[J]. Proc Natl Acad Sci USA, 2008, 105(42):16314-16319.DOI:10.1073/pnas.0808517105.
NAGELKERKE A, BUSSINK J, MUJCIC H, et al. Hypoxia stimulates migration of breast cancer cells via the PERK/ATF4/LAMP3-arm of the unfolded protein response[J]. Breast Cancer Res, 2013, 15(1):R2.DOI:10.1186/bcr3373.
MUJCIC H, RZYMSKI T, ROUSCHOP K M, et al. Hypoxic activation of the unfolded protein response (UPR) induces expression of the metastasis-associated gene LAMP3[J]. Radiother Oncol, 2009, 92(3):450-459.DOI:10.1016/j.radonc.2009.08.017.
YIN F, SHAREN G, YUAN F, et al. TIP30 regulates lipid metabolism in hepatocellular carcinoma by regulating SREBP1 through the Akt/mTOR signaling pathway[J]. Oncogenesis, 2017, 6(6):e347.DOI:10.1038/oncsis.2017.49.
JIA Y, WU C, KIM J, et al. Astaxanthin reduces hepatic lipid accumulations in high-fat-fed C57BL/6J mice via activation of peroxisome proliferator-activated receptor (PPAR) alpha and inhibition of PPAR gamma and Akt[J]. J Nutr Biochemi, 2016, 28:9-18.DOI:10.1016/j.jnutbio.2015.09.015.
LI J, HUANG Q, LONG X, et al. CD147 reprograms fatty acid metabolism in hepatocellular carcinoma cells through Akt/mTOR/SREBP1c and P38/PPARalpha pathways[J]. J Hepatol, 2015, 63(6):1378-1389.DOI:10.1016/j.jhep.2015.07.039.
LAPLANTE M, SABATINI D M. An emerging role of mTOR in lipid biosynthesis[J]. Cur Biol, 2009, 19(22):R1046-R52.DOI:10.1016/j.cub.2009.09.058.
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