脂肪酸结合蛋白4对BCG诱导巨噬细胞自噬的调控作用

doi:10.11843/j.issn.0366-6964.2020.09.024

畜牧兽医学报 ›› 2020, Vol. 51 ›› Issue (9): 2265-2274.doi: 10.11843/j.issn.0366-6964.2020.09.024

脂肪酸结合蛋白4对BCG诱导巨噬细胞自噬的调控作用

于嘉霖^1,2, 徐雅楠^1,2, 韩璐^1,2, 马沁梅^1,2, 吴晓玲^1,2, 邓光存^1,2*

1. 西部特色资源保护与利用教育部重点实验室, 银川 750021;
2. 宁夏大学生命科学学院, 银川 750021

收稿日期:2020-04-13 出版日期:2020-09-25 发布日期:2020-09-25
通讯作者: 邓光存,主要从事病原生物学研究,E-mail:dgc@nxu.edu.cn
作者简介:于嘉霖(1993-),男,宁夏吴忠人,硕士,主要从事病原生物学研究,E-mail:aries6687@163.com
基金资助:
国家自然科学基金（31760326）；宁夏重点研发计划项目（2018BFH03017）；西部一流学科建设重大创新项目（ZKZD2017001）；宁夏科技创新领军人才培养项目（KJT2017002）；宁夏大学研究生创新项目（GIP2019067）

Role of Fatty Acid Binding Protein 4 in Regulating Macrophage Autophagy Induced by BCG Infection

YU Jialin^1,2, XU Yanan^1,2, HAN Lu^1,2, MA Qinmei^1,2, WU Xiaoling^1,2, DENG Guangcun^1,2*

1. Key Lab of Ministry of Education for Protection and Utilization of Special Biological Rsources in Wstern China, Ningxia University, Yinchuan 750021, China;
2. College of Life Science, Ningxia University, Yinchuan 750021, China

Received:2020-04-13 Online:2020-09-25 Published:2020-09-25

摘要/Abstract

摘要： 旨在探讨脂肪酸结合蛋白4（fatty acid binding protein，FABP4）在牛分枝杆菌卡介苗（BCG）感染的巨噬细胞中对脂肪酸代谢与自噬的调控作用。本研究利用小干扰RNA技术敲减小鼠巨噬细胞系RAW264.7中FABP4的表达，并结合BCG感染，采用免疫印记和免疫荧光等技术，检测了FABP4蛋白表达、脂肪酸累积、脂肪酸β氧化和自噬相关蛋白表达等指标。结果表明，BCG感染极显著上调了小鼠巨噬细胞RAW264.7中FABP4的表达（P<0.001）并促进了脂肪酸的累积。FABP4小干扰RNA（siRNA-FABP4）能显著降低BCG感染后巨噬细胞中脂肪酸含量，伴随着肉毒碱棕榈酰移位酶1A（CPT1A）的表达上调（P<0.05），与ATP产量的提升（P<0.05）。同时，siRNA-FABP4极显著下调了自噬调控关键因子AMPK及自噬相关蛋白p-ULK1、ATG5、ATG7、ATG12和LC3B的表达（P<0.01）。以上研究结果表明，在BCG感染RAW264.7细胞过程中，下调了FABP4的表达，促进了脂肪酸的氧化，减少了巨噬细胞内脂肪酸的含量，并通过AMPK信号通路抑制了细胞自噬的发生。

关键词: FABP4, BCG, 巨噬细胞RAW264.7, 脂肪酸代谢, 细胞自噬

Abstract: To investigate the role of FABP4 on regulating fatty acid metabolism and autophagy in macrophage infected with BCG, small interfering RNA for FABP4 were transfected into RAW264.7 cells alone or combined with BCG infection. The immunoblot and immunofluorescence were used to detect the expression of FABP4, the accumulation of lipid droplets, the expression of fatty acid β-oxidation and autophagy related factors. The results showed that the infection of BCG increased the expression of FABP4 and accompanied with the accumulation of lipid droplets in RAW264.7. Moreover, the knockdown of FABP4 decreased lipid droplets but promoted the expression of carnitine palmitoyltransferase 1A (CPT1A) (P <0.05) and the production of ATP (P <0.05). Meanwhile, the knockdown of FABP4 suppressed the expression of AMPK, p-ULK1, ATG5, ATG7, ATG12 and LC3B which are autophagy related factors (P<0.001). Our results indicated that the knockdown of FABP4 promoted fatty acid oxidation, decreased fatty acid level and suppressed autophagy via AMPK signal pathway in BCG infected RAW264.7 cells.

Key words: FABP4, BCG, macrophage RAW264.7, fatty acid metabolism, autophagy

中图分类号:

S855.2

于嘉霖, 徐雅楠, 韩璐, 马沁梅, 吴晓玲, 邓光存. 脂肪酸结合蛋白4对BCG诱导巨噬细胞自噬的调控作用[J]. 畜牧兽医学报, 2020, 51(9): 2265-2274.

YU Jialin, XU Yanan, HAN Lu, MA Qinmei, WU Xiaoling, DENG Guangcun. Role of Fatty Acid Binding Protein 4 in Regulating Macrophage Autophagy Induced by BCG Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(9): 2265-2274.

参考文献 32

[1]	World Health Organization. Global tuberculosis report 2019[R]. Geneva, Switzerland:World Health Organization, 2019.
[2]	SHI L B, JIANG Q K, BUSHKIN Y, et al. Biphasic dynamics of macrophage immunometabolism during Mycobacterium tuberculosis infection[J]. mBio, 2019, 10(2):e02550-18.
[3]	HMAMA Z, PEÑA-DÍAZ S, JOSEPH S, et al. Immunoevasion and immunosuppression of the macrophage by Mycobacterium tuberculosis[J]. Immunol Rev, 2015, 264(1):220-232.
[4]	PAIK S, KIM J K, CHUNG C, et al. Autophagy:a new strategy for host-directed therapy of tuberculosis[J]. Virulence, 2019, 10(1):448-459.
[5]	PADHI A, PATTNAIK K, BISWAS M, et al. Mycobacterium tuberculosis LprE suppresses TLR2-dependent cathelicidin and autophagy expression to enhance bacterial survival in macrophages[J]. J Immunol, 2019, 203(10):2665-2678.
[6]	LEE H J, KO H J, SONG D K, et al. Lysophosphatidylcholine promotes phagosome maturation and regulates inflammatory mediator production through the protein kinase a-phosphatidylinositol 3 kinase-p38 mitogen-activated protein kinase signaling pathway during Mycobacterium tuberculosis infection in mouse macrophages[J]. Front Immunol, 2018, 9:920.
[7]	CUMMING B M, ADDICOTT K W, ADAMSON J H, et al. Mycobacterium tuberculosis induces decelerated bioenergetic metabolism in human macrophages[J]. eLife, 2018, 7:e39169.
[8]	AHLUWALIA P K, PANDEY R K, SEHAJPAL P K, et al. Perturbed microRNA expression by Mycobacterium tuberculosis promotes macrophage polarization leading to pro-survival foam cell[J]. Front Immunol, 2017, 8:107.
[9]	SINGH R, KAUSHIK S, WANG Y J, et al. Autophagy regulates lipid metabolism[J]. Nature, 2009, 458(7242):1131-1135.
[10]	KE R, XU Q C, LI C, et al. Mechanisms of AMPK in the maintenance of ATP balance during energy metabolism[J]. Cell Biol Int, 2018, 42(4):384-392.
[11]	MEI S, NI H M, MANLEY S, et al. Differential roles of unsaturated and saturated fatty acids on autophagy and apoptosis in hepatocytes[J]. J Pharmacol Exp Ther, 2011, 339(2):487-498.
[12]	FURUHASHI M, SAITOH S, SHIMAMOTO K, et al. Fatty acid-binding protein 4 (FABP4):pathophysiological insights and potent clinical biomarker of metabolic and cardiovascular diseases[J]. Clin Med Insights Cardiol, 2014, 8(S3):23-33.
[13]	GUAITA-ESTERUELAS S, BOSQUET A, SAAVEDRA P, et al. Exogenous FABP4 increases breast cancer cell proliferation and activates the expression of fatty acid transport proteins[J]. Mol Carcinogen, 2017, 56(1):208-217.
[14]	BOß M, KEMMERER M, BRÜNE B, et al. FABP4 inhibition suppresses PPARγ activity and VLDL-induced foam cell formation in IL-4-polarized human macrophages[J]. Atherosclerosis, 2015, 240(2):424-430.
[15]	DENG T M, WANG Y H, WANG C C, et al. FABP4 silencing ameliorates hypoxia reoxygenation injury through the attenuation of endoplasmic reticulum stress-mediated apoptosis by activating PI3K/Akt pathway[J]. Life Sci, 2019, 224:149-156.
[16]	ZHANG Q W, SUN J X, WANG Y L, et al. Antimycobacterial and anti-inflammatory mechanisms of baicalin via induced autophagy in macrophages infected with Mycobacterium tuberculosis[J]. Front Microbiol, 2017, 8:2142.
[17]	WU X L, ZHANG J M, MA C J, et al. A role for Wnt/β-catenin signalling in suppressing bacillus calmette-guerin-induced macrophage autophagy[J]. Microb Pathog, 2019, 127:277-287.
[18]	马沁梅, 韩璐, 邓光存, 等. 自噬在结核分枝杆菌感染中作用的研究进展[J]. 中国细胞生物学学报, 2019, 41(9):1797-1804.MA Q M, HAN L, DENG G C, et al. The research progress of autophagy in Mycobacterium tuberculosis infection[J]. Chinese Journal of Cell Biology, 2019, 41(9):1797-1804. (in Chinese)
[19]	PAHARI S, NEGI S, AQDAS M, et al. Induction of autophagy through CLEC4E in combination with TLR4:an innovative strategy to restrict the survival of Mycobacterium tuberculosis[J]. Autophagy, 2020, 16(6):1021-1043.
[20]	KIM J K, LEE H M, PARK K S, et al. MIR144* inhibits antimicrobial responses against Mycobacterium tuberculosis in human monocytes and macrophages by targeting the autophagy protein DRAM2[J]. Autophagy, 2017, 13(2):423-441.
[21]	KIM M J, WAINWRIGHT H C, LOCKETZ M, et al. Caseation of human tuberculosis granulomas correlates with elevated host lipid metabolism[J]. Embo Mol Med, 2010, 2(7):258-274.
[22]	REMMERIE A, SCOTT C L. Macrophages and lipid metabolism[J]. Cell Immunol, 2018, 330:27-42.
[23]	LEE W, VANDERVEN B C, FAHEY R J, et al. Intracellular Mycobacterium tuberculosis exploits host-derived fatty acids to limit metabolic stress[J]. J Biol Chem, 2013, 288(10):6788-6800.
[24]	HOTAMISLIGIL G S, BERNLOHR D A. Metabolic functions of FABPs-mechanisms and therapeutic implications[J]. Nat Rev Endocrinol, 2015, 11(10):592-605.
[25]	HARJES U, BRIDGES E, GHARPURE K M, et al. Antiangiogenic and tumour inhibitory effects of downregulating tumour endothelial FABP4[J]. Oncogene, 2017, 36(7):912-921.
[26]	HERAS-SANDOVAL D, PÉREZ-ROJAS J M, HERNÁNDEZ-DAMIÁN J, et al. The role of PI3K/AKT/mTOR pathway in the modulation of autophagy and the clearance of protein aggregates in neurodegeneration[J]. Cell Signal, 2014, 26(12):2694-2701.
[27]	KHAWAR M B, GAO H, LI W. Autophagy and lipid metabolism[M]//QIN Z H. Autophagy:Biology and Diseases. Singapore:Springer, 2019:359-374.
[28]	DUAN W J, LI Y F, LIU F L, et al. A SIRT3/AMPK/autophagy network orchestrates the protective effects of trans-resveratrol in stressed peritoneal macrophages and RAW 264.7 macrophages[J]. Free Radical Biol Med, 2016, 95:230-242.
[29]	LIN S C, HARDIE C. AMPK:sensing glucose as well as cellular energy status[J]. Cell Metab, 2018, 27(2):299-313.
[30]	EGAN D, KIM J, SHAW R J, et al. The autophagy initiating kinase ULK1 is regulated via opposing phosphorylation by AMPK and mTOR[J]. Autophagy, 2011, 7(6):643-644.
[31]	ZACHARI M, GANLEY I G. The mammalian ULK1 complex and autophagy initiation[J]. Essays Biochem, 2017, 61(6):585-596.
[32]	LI W, ZHANG L L. Regulation of ATG and autophagy initiation[M]//QIN Z H. Autophagy:Biology and Diseases. Singapore:Springer, 2019:41-65.

脂肪酸结合蛋白4对BCG诱导巨噬细胞自噬的调控作用

Role of Fatty Acid Binding Protein 4 in Regulating Macrophage Autophagy Induced by BCG Infection

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献 32

相关文章 15

编辑推荐

Metrics

本文评价

[1]	刘悦阳, 李梦媛, 聂雪伊, 马亚博, 侯雨欣, 马伯利, 杨易, 徐金瑞. 钙结合蛋白S100A4对BCG感染THP-1细胞自噬的调控作用[J]. 畜牧兽医学报, 2024, 55(1): 311-322.
[2]	安琪, 于嘉霖, 吴晓玲, 邓光存. 谷氨酰胺对BCG诱导小鼠传代巨噬细胞凋亡的调控作用[J]. 畜牧兽医学报, 2023, 54(7): 3054-3063.
[3]	王崇年, 于嘉霖, 宫照乾, 吴晓玲, 邓光存. 脂肪分化相关蛋白2对BCG诱导小鼠传代巨噬细胞自噬的调控作用[J]. 畜牧兽医学报, 2023, 54(5): 2134-2146.
[4]	王立斌, 王萌, 孙莹, 陈睿, 张甜甜, 黄振华, 张倩, 余四九, 潘阳阳. CYP19A1调控内源性雌激素合成促进牦牛COCs细胞自噬和早期发育能力[J]. 畜牧兽医学报, 2022, 53(12): 4283-4295.
[5]	陈琪, 杜长征, 郑雪迪, 马伯利, 徐金瑞, 杨易. Wnt5a介导Wnt/Ca²⁺通路对BCG诱导牛原代肺泡上皮细胞自噬的调控作用[J]. 畜牧兽医学报, 2022, 53(11): 4071-4080.
[6]	王正荣, 马勋, 张艳艳, 孟季蒙, 薄新文. 基于转录组测序分析细粒棘球绦虫原头蚴与巨噬细胞RAW264.7免疫互作相关基因[J]. 畜牧兽医学报, 2022, 53(11): 3975-3988.
[7]	王正荣, 马勋, 张艳艳, 孟季蒙, 薄新文. Th1/Th2型免疫反应相关基因在巨噬细胞RAW264.7应对细粒棘球绦虫原头蚴刺激时的表达谱研究[J]. 畜牧兽医学报, 2022, 53(1): 250-262.
[8]	骆佳, 徐金瑞, 李武, 王玉炯. BCG诱导RAW264.7细胞脂肪酸氧化对细胞自噬和促炎因子表达的调控作用[J]. 畜牧兽医学报, 2021, 52(9): 2617-2625.
[9]	贾艳艳, 赵莹莹, 余祖华, 何雷, 廖成水, 李静, 郁川, 张春杰, 李银聚. Beclin-1基因shRNA慢病毒载体的构建及其对B16F10细胞自噬及活力的影响[J]. 畜牧兽医学报, 2021, 52(9): 2609-2616.
[10]	方舒, 张嘉美, 杨易, 韩璐, 马臣杰, 吴晓玲, 邓光存. 受体相互作用蛋白1(RIP1)对BCG诱导小鼠巨噬细胞凋亡的调控作用[J]. 畜牧兽医学报, 2019, 50(3): 645-653.
[11]	罗永莉, 江艾耘, 胡志刚, 李相兰, 杨晓迪, 旷策嫣, 周荣琼. RNA干扰abcg-5基因对犬弓首蛔虫感染性的影响[J]. 畜牧兽医学报, 2019, 50(2): 390-397.
[12]	胡林, 南良康, 张敏, 许苏童, 杨了寒, 李帅峰, 王凯, 刘正飞, 张淑君. 猪Beclin1基因调控伪狂犬病病毒诱导细胞自噬和凋亡的机制分析[J]. 畜牧兽医学报, 2018, 49(6): 1274-1281.
[13]	任昊, 李丽芳, 王彦美, 黄楠, 裴芳樱, 李嘉威, 孙耀贵, 段智变, 李宏全, 王文魁. 缺氧条件下巨噬细胞移动抑制因子对肉鸡心肌脂肪酸代谢的影响[J]. 畜牧兽医学报, 2018, 49(1): 195-202.
[14]	杨了寒, 王凯, 许苏童, 张敏, 胡林, 何启盖, 张淑君. P58IPK通过PERK/eIF2α通路调控猪圆环病毒2型所诱导的细胞自噬[J]. 畜牧兽医学报, 2017, 48(11): 2125-2134.
[15]	宫晓炜，郑福英，陈启伟，刘永生，李兆才，周继章，才学鹏，殷宏. 细胞自噬对牛病毒性腹泻病毒复制的影响[J]. 畜牧兽医学报, 2015, 46(7): 1201-1207.