油酸上调围脂滴蛋白2表达增强巨噬细胞对结核分枝杆菌的清除作用

doi:10.19982/j.issn.1000-6621.20240489

中国防痨杂志 ›› 2025, Vol. 47 ›› Issue (1): 72-76.doi: 10.19982/j.issn.1000-6621.20240489

油酸上调围脂滴蛋白2表达增强巨噬细胞对结核分枝杆菌的清除作用

卢海林¹^,², 汪文斐², 陶文慧², 林培翀², 陈心春³, 邓国防⁴(), 谢水祥¹()

¹赣南医科大学基础医学院,赣州 341000
²深圳国家感染性疾病临床医学研究中心,深圳 518000
³深圳大学医学部,深圳 518000
⁴深圳市第三人民医院肺病二科,深圳 518000

收稿日期:2024-11-04 出版日期:2025-01-10 发布日期:2025-01-02
通信作者: 邓国防,谢水祥 E-mail:jxxk1035@yeah.net;xsxw2002@163.com
基金资助:
深圳市第三人民医院研究基金(G202205);深圳市第三人民医院研究基金(G2022003);深圳市第三人民医院研究基金(G2022155);深圳市第三人民医院研究基金(24250G1025)

Oleic acid upregulates the expression of perilipin 2 enhancing macrophage clearance of Mycobacterium tuberculosis

Lu Hailin¹^,², Wang Wenfei², Tao Wenhui², Lin Peicong², Chen Xinchun³, Deng Guofang⁴(), Xie Shuixiang¹()

¹School of Basic Medicine, Gannan Medical University, Ganzhou 341000, China
²National Clinical Research Center for Infectious Diseases, Shenzhen 518000, China
³Shenzhen University School of Medicine, Shenzhen 518000, China
⁴Division Two of Pulmonary Diseases Department, The Third People’s Hospital of Shenzhen, Shenzhen 518000, China

Received:2024-11-04 Online:2025-01-10 Published:2025-01-02
Contact: Deng Guofang,Xie Shuixiang E-mail:jxxk1035@yeah.net;xsxw2002@163.com
Supported by:
Shenzhen Third People’s Hospital Research Foundation(G202205);Shenzhen Third People’s Hospital Research Foundation(G2022003);Shenzhen Third People’s Hospital Research Foundation(G2022155);Shenzhen Third People’s Hospital Research Foundation(24250G1025)

摘要/Abstract

摘要：

目的：研究油酸对巨噬细胞清除结核分枝杆菌(Mycobacterium tuberculosis,MTB)的作用,并进一步探讨其作用机制。方法：使用人单核细胞白血病细胞(THP-1)细胞系诱导的巨噬细胞,加入或不加入油酸(oleic acid,OA)后感染MTB标准株H37Ra,用菌落形成单位(colony-forming unit,CFU)计数及免疫荧光方式检测油酸对巨噬细胞清除MTB的影响。通过RNA-seq筛选出关键靶基因,并采用基因富集分析筛选出相关通路,实时荧光定量PCR验证基因表达。结果：MTB标准株H37Ra感染未经油酸诱导(H37Ra组)巨噬细胞后72h的菌落计数单位(CFU)中位数(四分位数)为[49×10⁴(48×10⁴,59×10⁴)],明显高于经油酸诱导(OA+H37Ra组)的巨噬细胞的CFU[34×10⁴(30×10⁴,42×10⁴)],差异有统计学意义(U=3.500,P=0.017)。RNA-Seq数据显示,围脂滴蛋白2(recombinant perilipin 2,PLIN2)的表达在OA+H37Ra组明显高于H37Ra组。在油酸诱导的巨噬细胞中,敲低PLIN2基因后的CFU结果显示,在H37Ra感染72h后,敲低PLIN2组的CFU中位数(四分位数)为86×10⁴(78×10⁴,96×10⁴),明显高于对照空载体组[56×10⁴(54×10⁴,62×10⁴)],差异有统计学意义(U=3.000,P=0.015)。OA+H37Ra组中,PLIN2的相对表达量为3.219±0.298,明显高于H37Ra组的0.555±0.028,差异有统计学意义(t=15.410,P<0.01);OA+H37Ra组中PPARγ的相对表达量为0.666±0.075,明显低于H37Ra组的2.217±0.153,差异有统计学意义(t=14.700, P<0.01);OA+H37Ra组中,ACOX1、HADHA基因的相对表达量分别为1.410±0.124和1.107±0.111,均明显低于H37Ra组的2.476±0.207和3.140±0.240,差异均有统计学意义(t=13.520,P<0.01;t=12.760,P<0.01)。结论：油酸通过脂滴表面蛋白PLIN2调控巨噬细胞的脂肪酸氧化促进巨噬细胞对MTB的清除。

关键词: 分枝杆菌,结核, 巨噬细胞, 油酸, 围脂滴蛋白, 基因表达调控,细菌

Abstract:

Objective: To investigate the role of oleic acid (OA) in the clearance of Mycobacterium tuberculosis (MTB) by macrophages and explore the underlying mechanisms. Methods: Macrophages induced from the human monocyte leukemia cell line (THP-1) were infected with the standard MTB strain H37Ra with or without oleic acid. The effect of oleic acid on the clearance of MTB by macrophages was assessed using colony-forming unit (CFU). Key target genes were identified through RNA-seq, and relevant pathways were determined using gene enrichment analysis. Real-time quantitative PCR was used for validating gene expression. Results: After 72 hours of infection with H37Ra, the CFU for macrophages (H37Ra group) was 49×10⁴ (48×10⁴, 59×10⁴), significantly higher than that of macrophages induced with oleic acid (OA+H37Ra group) at 34×10⁴ (30×10⁴, 42×10⁴) (U=3.500, P=0.017). RNA-seq data showed that the expression of recombinant perilipin 2 (PLIN2) in the OA+H37Ra group was significantly higher than in the H37Ra group. In oleic acid-induced macrophages, the CFU in the PLIN2 knock down group was 86×10⁴ (78×10⁴, 96×10⁴), significantly higher than that in the control group (56×10⁴ (54×10⁴, 62×10⁴); U=3.000, P=0.015). In the OA+H37Ra group, the relative expression of PLIN2 was 3.219±0.298, significantly higher than the 0.555±0.028 in the H37Ra group (t=15.410, P<0.01). The relative expression of PPARγ in the OA+H37Ra group was 0.666±0.075, significantly lower than the 2.217±0.153 in the H37Ra group (t=14.700, P<0.01). In the OA+H37Ra group, the relative expression of ACOX1 and HADHA was 1.410±0.124 and 1.107±0.111, both significantly lower than the H37Ra group of 2.476±0.207 and 3.140±0.240 (t=13.520, P<0.01; t=12.760, P<0.01) respectively. Conclusion: Oleic acid regulates fatty acid oxidation in macrophages through the lipid droplet surface protein PLIN2, promoting the clearance of MTB by macrophages.

Key words: Mycobacterium tuberculosis, Macrophages, Oleic acid, Perilipin, Gene expression regulation, bacterial

中图分类号:

R378.91

卢海林, 汪文斐, 陶文慧, 林培翀, 陈心春, 邓国防, 谢水祥. 油酸上调围脂滴蛋白2表达增强巨噬细胞对结核分枝杆菌的清除作用[J]. 中国防痨杂志, 2025, 47(1): 72-76. doi: 10.19982/j.issn.1000-6621.20240489

Lu Hailin, Wang Wenfei, Tao Wenhui, Lin Peicong, Chen Xinchun, Deng Guofang, Xie Shuixiang. Oleic acid upregulates the expression of perilipin 2 enhancing macrophage clearance of Mycobacterium tuberculosis[J]. Chinese Journal of Antituberculosis, 2025, 47(1): 72-76. doi: 10.19982/j.issn.1000-6621.20240489

图/表 1

参考文献 19

[1]	Zhao L, Fan K, Sun X, et al. Host-directed therapy against mycobacterium tuberculosis infections with diabetes mellitus. Front Immunol, 2023, 14: 1305325. doi:10.3389/fimmu.2023.1305325.
[2]	Jeong EK, Lee HJ, Jung YJ. Host-Directed Therapies for Tuberculosis. Pathogens, 2022, 11(11):1291. doi:10.3390/pathogens11111291.
[3]	Kilinç G, Saris A, Ottenhoff THM, et al. Host-directed therapy to combat mycobacterial infections. Immunol Rev, 2021, 301(1): 62-83. doi:10.1111/imr.12951. URL pmid: 33565103
[4]	Liu CH, Liu H, Ge B. Innate immunity in tuberculosis: host defense vs pathogen evasion. Cell Mol Immunol, 2017, 14(12): 963-975. doi:10.1038/cmi.2017.88.
[5]	Wu Y, Chen K, Li L, et al. Plin2-mediated lipid droplet mobilization accelerates exit from pluripotency by lipidomic remodeling and histone acetylation. Cell Death Differ, 2022, 29(11): 2316-2331. doi:10.1038/s41418-022-01018-8. URL pmid: 35614132
[6]	Bosch M, Pol A. Eukaryotic lipid droplets: metabolic hubs, and immune first responders. Trends Endocrinol Metab, 2022, 33(3): 218-229. doi:10.1016/j.tem.2021.12.006.
[7]	Apte MS, Joshi AS. Membrane shaping proteins, lipids, and cytoskeleton: Recipe for nascent lipid droplet formation. Bioessays, 2022, 44(9): e2200038. doi:10.1002/bies.202200038.
[8]	Laval T, Chaumont L, Demangel C. Not too fat to fight: The emerging role of macrophage fatty acid metabolism in immunity to Mycobacterium tuberculosis. Immunol Rev, 2021, 301(1): 84-97. doi:10.1111/imr.12952.
[9]	Herker E, Vieyres G, Beller M, et al. Lipid Droplet Contact Sites in Health and Disease. Trends Cell Biol, 2021, 31(5):345-358. doi:10.1016/j.tcb.2021.01.004.
[10]	Bosch M, Sánchez-Álvarez M, Fajardo A, et al. Mammalian lipid droplets are innate immune hubs integrating cell metabolism and host defense. Science, 2020, 370(6514): eaay8085. doi:10.1126/science.aay8085.
[11]	Agarwal P, Combes TW, Shojaee-Moradie F, et al. Foam Cells Control Mycobacterium tuberculosis Infection. Front Microbiol, 2020, 11:1394. doi:10.3389/fmicb.2020.01394. URL pmid: 32754123
[12]	Olzmann JA, Carvalho P. Dynamics and functions of lipid droplets. Nat Rev Mol Cell Biol, 2019, 20(3):137-155. doi:10.1038/s41580-018-0085-z.
[13]	Jarc E, Petan T. Lipid Droplets and the Management of Cellular Stress. Yale J Biol Med, 2019, 92(3):435-452. pmid: 31543707
[14]	Howard NC, Khader SA. Immunometabolism during Mycobacterium tuberculosis Infection. Trends Microbiol, 2020, 28(10): 832-850. doi:10.1016/j.tim.2020.04.010.
[15]	Sheedy FJ, Divangahi M. Targeting immunometabolism in host defence against Mycobacterium tuberculosis. Immunology, 2021, 162(2):145-159. doi:10.1111/imm.13276.
[16]	Escoll P, Buchrieser C. Metabolic reprogramming: an innate cellular defence mechanism against intracellular bacteria?. Curr Opin Immunol, 2019, 60: 117-123. doi:10.1016/j.coi.2019.05.009. URL pmid: 31247377
[17]	Kewcharoenwong C, Saenwongsa W, Willcocks SJ, et al. Glibenclamide alters interleukin-8 and interleukin-1β of primary human monocytes from diabetes patients against Mycobacterium tuberculosis infection. Tuberculosis (Edinb), 2020, 123:101939. doi:10.1016/j.tube.2020.101939.
[18]	Knight M, Braverman J, Asfaha K, et al. Lipid droplet formation in Mycobacterium tuberculosis infected macrophages requires IFN-γ/HIF-1α signaling and supports host defense. PLoS Pathog, 2018, 14(1): e1006874. doi:10.1371/journal.ppat.1006874.
[19]	Arnett E, Weaver AM, Woodyard KC, et al. PPARγ is critical for Mycobacterium tuberculosis induction of Mcl-1 and limitation of human macrophage apoptosis. PLoS Pathog, 2018, 14(6): e1007100. doi:10.1371/journal.ppat.1007100.

组别	感染6h(×10⁴)	感染72h(×10⁴)
H37Ra组	24(17,32)	49(48,59)
OA+H37Ra组	22(21,23)	34(30,42)
U值	0.379	3.500
P值	0.998	0.017

油酸上调围脂滴蛋白2表达增强巨噬细胞对结核分枝杆菌的清除作用

Oleic acid upregulates the expression of perilipin 2 enhancing macrophage clearance of Mycobacterium tuberculosis

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 1

参考文献 19

相关文章 15

编辑推荐

Metrics

本文评价

友情链接

[1]	赵艳梅, 韩悦, 柯云玲, 庄让笑. 巨噬细胞极化在肺部疾病中的作用机制[J]. 中国防痨杂志, 2024, 46(S2): 22-24.
[2]	王艺霖, 吴潇, 逄宇, 李姗姗. 奥布替尼在分枝杆菌感染宿主巨噬细胞中的免疫调控作用[J]. 中国防痨杂志, 2024, 46(9): 1063-1068.
[3]	刘浩瀚, 马子淳, 逄宇, 李姗姗. G蛋白偶联受体在宿主抗结核分枝杆菌感染中的功能及作用机制[J]. 中国防痨杂志, 2024, 46(7): 839-844.
[4]	陈飞飞, 郑永智, 吴素芳, 康乾, 晋春阳. 活性氧/蛋白激酶RNA样内质网激酶信号轴对卡介苗诱导RAW264.7小鼠巨噬细胞铁死亡的调控作用[J]. 中国防痨杂志, 2024, 46(12): 1448-1458.
[5]	赵勇, 梁丽丽, 江南. LncRNA GAS5靶向调控miR-144-3p对结核分枝杆菌感染的巨噬细胞凋亡和炎性反应的影响[J]. 中国防痨杂志, 2023, 45(8): 744-751.
[6]	何萍, 王怡婷, 宋泽萱, 夏辉, 王胜芬, 贺文从, 郑扬, 赵雁林. 结核分枝杆菌Rv2333c在巨噬细胞内功能的初步研究[J]. 中国防痨杂志, 2023, 45(6): 566-574.
[7]	史雨婷, 董静, 贾红彦, 朱传智, 杨斌, 李自慧, 孙琦, 杜博平, 邢爱英, 张宗德, 潘丽萍. miR-99a-5p在结核分枝杆菌感染宿主巨噬细胞中的免疫调控作用[J]. 中国防痨杂志, 2023, 45(5): 464-471.
[8]	段玉衡, 张蓝月, 董静, 史雨婷, 贾红彦, 李自慧, 邢爱英, 杜博平, 孙琦, 潘丽萍, 朱传智, 张宗德. 结核分枝杆菌乙酰转移酶fadA3对宿主蛋白乙酰化修饰及其体内存活影响的研究[J]. 中国防痨杂志, 2023, 45(4): 391-400.
[9]	吴显劲, 黄海勇, 萧乐瑶, 徐军发. 巨噬细胞极化与结核分枝杆菌感染的研究进展[J]. 中国防痨杂志, 2023, 45(12): 1198-1204.
[10]	南岳, 龙美贞, 董玉慧, 王元智, 周向梅. 巨噬细胞糖代谢重编程在结核分枝杆菌感染中的研究进展[J]. 中国防痨杂志, 2023, 45(11): 1097-1102.
[11]	李银虹, 刘芳琳, 鹿振辉, 姜昕. 冬凌草甲素抗结核病理损伤的作用机制研究[J]. 中国防痨杂志, 2022, 44(8): 849-854.
[12]	吴托雅, 石金, 郭继东, 刘原园, 逄宇, 鲁洁, 高飞. miR-21-3p调节结核分枝杆菌在宿主巨噬细胞内存活机制的研究[J]. 中国防痨杂志, 2021, 43(5): 475-481.
[13]	向海滨,梁求真,李新霞,宋兴华. 巨噬细胞的抗结核纳米递药系统的应用进展[J]. 中国防痨杂志, 2020, 42(4): 398-403.
[14]	孙锦霞,张晴雯,李银虹,姜昕. 杨梅素通过磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路诱导MTB感染巨噬细胞发生自噬的研究[J]. 中国防痨杂志, 2020, 42(2): 101-107.
[15]	陈曦,刘忠泉,王彬,朱慧,付雷,李媛媛,陆宇. 14种抗结核药物在巨噬细胞内的抗结核活性评价[J]. 中国防痨杂志, 2019, 41(9): 993-998.