Sortilin调控ABCA1蛋白水平影响荷脂巨噬细胞内脂质流出

doi:10.13418/j.issn.1001-165x.2020.05.015

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (5): 568-573.doi: 10.13418/j.issn.1001-165x.2020.05.015

Sortilin调控ABCA1蛋白水平影响荷脂巨噬细胞内脂质流出

钟丽园¹，　张明鑫²，　吕运成²，　陶水英²，　胡杨²，　王茜²，　唐志桦²，　彭田红²

1.南华大学肿瘤研究所湖南省肿瘤细胞与分子病理学重点实验室湖南省胃癌研究中心，湖南衡阳 421000；
2.南华大学衡阳医学院应用解剖与生殖医学研究所，湖南衡阳 421000

收稿日期:2019-10-31 出版日期:2020-09-25 发布日期:2020-10-21
通讯作者: 彭田红，教授，E-mail：thpeng67@163.com
作者简介:钟丽园（1992-），女，湖南浏阳人，助教，硕士，研究方向为动脉粥样硬化的病因及发病学： E-mail：2257868339@qq.com；共同第一作者：张明鑫(1998-)，男，湖南耒阳人，本科在读，研究方向为动脉粥样硬化的病因及发病学，E-mail：1553498778@qq.com
基金资助:
国家自然科学基金(81770460)；加拿大萨斯喀彻温省卫生研究基金会博士后基金(SHRF4144)；衡阳市科学技术发展计划项目（2017KJ268、2017KJ182）；南华大学博士启动基金(2014XQD37)；大学生研究性学习和创新性实验计划项目（201810555015）：湖南省自然科学基金（2020JJ4532)

Sortilin alters lipid efflux from lipid-laden THP-1 macrophage by regulating ABCA1 protein

ZHONG Li-yuan¹, ZHANG Ming-xin², LV Yun-cheng², TAO Shui-ying², HU Yang², WANG Xi², TANG Zhi-hua², PENG Tian-hong²

1.Cancer Research Institute, University of South China, Hunan Provincial Key Laboratory of Cancer Cellular And MolecularPathology, Center for Gastric Cancer Research of Hunan Province, Hengyang 421000, Hunan Porvince, China; 2.Clinical Anatomy & Reproductive Medicine Applicaton Institute, Hengyang Medical College of University of South China, Hengyang 421000, Hunan Porvince, China

Received:2019-10-31 Online:2020-09-25 Published:2020-10-21

摘要/Abstract

摘要： 目的　探讨Sortilin对荷脂THP-1巨噬细胞ABCA1蛋白表达及胞内脂质流出的影响。方法　采用Western blot和qRT-PCR检测细胞内ABCA1蛋白和mRNA表达水平；Co-IP实验检测Sortilin和ABCA1的结合情况；高效液相色谱法检测细胞内总胆固醇、游离胆固醇、胆固醇酯含量变化；油红O染色观察细胞内脂滴情况。结果　Sortilin下调细胞内ABCA1蛋白水平，但对其mRNA水平无显著影响；Co-IP结果表明Sortilin能与ABCA1结合；与对照组相比，Sortilin过表达后荷脂巨噬细胞内胆固醇流出减少，胞内脂质含量增加且脂滴肥大，数量明显增多，Sortilin沉默后荷脂巨噬细胞胆固醇流出增加，胞内脂质含量减少，脂滴瘦小，数量减少；溶酶体抑制剂氯喹共处理可部分逆转Sortilin过表达对巨噬细胞ABCA1蛋白和胆固醇流出的抑制作用。结论　Sortilin下调巨噬细胞ABCA1蛋白水平，抑制胆固醇流出，促进胞内脂质蓄积。

关键词: Sortilin, 　巨噬细胞, 　ABCA1, 　胆固醇流出

Abstract: Objective　To investigate the effect of Sortilin on protein level of macrophage ABCA1 and intracellular lipid efflux. Methods The protein and mRNA levels of lipid-laden macrophage ABCA1 were detected by western blot (WB) and quantitative real-time polymerase chain reaction (qRT-PCR) respectively; the combination of Sortilin with ABCA1 was detected by co-immunoprecipitation (Co-IP); macrophage cholesterol efflux was measured by liquid scintillation counting. Total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) were determined by high performance liquid chromatography (HPLC). Intracellular lipid droplets were observed by oil red O staining. Results Sortilin overexpression significantly decreased protein level of ABCA1, while Sortilin silence obviously raised cellular level of ABCA1 protein in lipid-laden macrophage. ABCA1 mRNA has no significant change in these three groups. Co-IP assay showed that Sortilin combined with ABCA1 protein. Compared to control group, Sortilin overexpression decreased cholesterol efflux from lipid-laden macrophage, resulting in macrophage cholesterol accumulation and lipid droplet generation. Sortilin silence dramatically enhanced cholesterol efflux and decreased lipid droplet formation. Co-treatment with chloroquine can partially abolish the inhibitory role of Sortilin overexpression in ABCA1 protein and lipid efflux of lipid-laden macrophage.　Conclusions　Sortilin reduces protein level of macrophage ABCA1 and intracellular cholesterol efflux, causes the accumulation of intracellular lipid.

Key words: Sortilin, 　Macrophage, 　ABCA1, 　Cholesterol efflux

中图分类号:

R329.28

钟丽园, 张明鑫, 吕运成, 陶水英, 胡杨, 王茜, 唐志桦, 彭田红. Sortilin调控ABCA1蛋白水平影响荷脂巨噬细胞内脂质流出[J]. 中国临床解剖学杂志, 2020, 38(5): 568-573.

ZHONG Li-yuan, ZHANG Ming-xin, LV Yun-cheng, TAO Shui-ying, HU Yang, WANG Xi, TANG Zhi-hua, PENG Tian-hong. Sortilin alters lipid efflux from lipid-laden THP-1 macrophage by regulating ABCA1 protein[J]. Chinese Journal of Clinical Anatomy, 2020, 38(5): 568-573.

参考文献 15

[1]	Gao AB, Cayabyab F.S, Chen X, et al. Implications of Sortilin in Lipid Metabolism and Lipid Disorder Diseases[J]. DNA and cell biology, 2017, 36(12): 1050-1061.
[2]	高安博, 彭田红, 钟丽园, 等. NF-κB调控Sortilin促进荷脂THP-1巨噬细胞泡沫化[J]. 中国动脉粥样硬化杂志, 2018,26(07):666-671.
[3]	钟丽园, 彭田红,高安博, 等. 分拣受体Sortilin促进巨噬细胞内脂质蓄积[J]. 中国动脉硬化杂志, 2018,26(02):139-143.
[4]	Xie CM, Lin L, Long Y, et al. ABCA1 affects placental function via trophoblast and macrophage[J]. Life sciences, 2017, 191: 150-156.
[5]	Hafiane A, Genest J. ATP binding cassette A1 (ABCA1) mediates microparticle formation during high-density lipoprotein (HDL) biogenesis[J]. Atherosclerosis, 2017, 257: 90-99.
[6]	Chistlkov DA, Bobryshev YV, Orekhov AN. Macrophage-mediated cholesterol handling in atherosclerosis[J]. J Cell Mol Med, 2016, 20(1): 17-28.
[7]	Lv YC, Yin K, Fu YC, et al. Posttranscriptional regulation of ATP-Binding cassette transporter A1 in lipid metabolism[J]. DNA and Cell Biology, 2013, 32(7): 348-358.
[8]	Reitz C, Tosto G, Vardrjan B, et al. Independent and epistatic effects of variants in VPS10-d receptors on Alzheimer disease risk and processing of the amyloid precursor protein (APP)[J]. Translational Psychiatry, 2013, 1-9.
[9]	Strong A, Patel K, Rader DJ. Sortilin and lipoprotein metabolism: making sense out of complexity[J]. Curr Opin Lipidol, 2014, 5(25): 350-357.
[10]	Conlon D. Role of sortilin in lipid metabolism[J]. urr Opin Lipido, 2019, 30(3): 198-204.
[11]	Zhong LY, Cayabyab F.S, Tang CK, et al. Sortilin: A novel regulator in lipid metabolism and atherogenesis[J]. Clinica chimica acta, 2016, 460(2016): 11-17.
[12]	Nilsson SK, Christensen S, Raarup MK, et al. Endocytosis of Apolipoprotein A-V by Members of the Low Density Lipoprotein Receptor and the Vps10p Domain Receptor Families[J]. Journal of Biological Chemistry, 2008, 283(38): 25920-25927.
[13]	Musunuru K, Strong A, Frank-kamenetsky M, et al. From noncoding variant to phenotype via SORT1 at the 1p13 cholesterol locus[J]. Nature, 2010, 466(7307): 714-719.
[14]	Nykjare A, LEE R, Teng KK, et al. Sortilin is essential for proNGF-induced neuronal cell death[J]. Nature, 2004, 427(6977): 843-848.
[15]	Bashore AC, Liu M, Kry CC, et al. Targeted Deletion of Hepatocyte Abca1 Increases Plasma HDL Reverse Cholesterol Transport via the LDL Receptor[J]. Arterioscler Thromb Vasc Biol, 2019, 39(9): 1747-1761.

Sortilin调控ABCA1蛋白水平影响荷脂巨噬细胞内脂质流出

Sortilin alters lipid efflux from lipid-laden THP-1 macrophage by regulating ABCA1 protein

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