脂肪酸结合蛋白4对卡介苗感染巨噬细胞趋化因子分泌的调控作用

doi:10.11843/j.issn.0366-6964.2025.07.040

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (7): 3484-3494.doi: 10.11843/j.issn.0366-6964.2025.07.040

脂肪酸结合蛋白4对卡介苗感染巨噬细胞趋化因子分泌的调控作用

赵燕萍¹^,²(), 王晓平³, 于嘉霖¹^,², 宫照乾¹^,², 吴晓玲¹^,², 邓光存¹^,²^,*()

1. 宁夏大学生命科学学院，银川 750021
2. 宁夏大学西部特色生物资源保护与利用教育部重点实验室，银川 750021
3. 宁夏回族自治区第四人民医院，银川 750021

收稿日期:2024-09-19 出版日期:2025-07-23 发布日期:2025-07-25
通讯作者: 邓光存 E-mail:3414153939@qq.com;dgc@nxu.edu.cn
作者简介:赵燕萍(1995-)，女，宁夏吴忠人，硕士生，主要从事动物病原生物学与宿主免疫相关研究，E-mail: 3414153939@qq.com
基金资助:
国家自然科学基金项目(32160162；32060160；32460172)；宁夏自然科学基金重点项目(2023AAC02015)；宁夏大学研究生创新项目(CXXM2024-23)

Role of Fatty Acid-Binding Protein 4 on Chemokine Secretion in Bacillus Calmette-Guérin Infected Macrophages

ZHAO Yanping¹^,²(), WANG Xiaoping³, YU Jialin¹^,², GONG Zhaoqian¹^,², WU Xiaoling¹^,², DENG Guangcun¹^,²^,*()

1. School of Life Sciences, Ningxia University, Yinchuan 750021, China
2. Key Laboratory of Ministry of Education for Protection and Utilization of Special Biological Resources in Western China, Ningxia University, Yinchuan 750021, China
3. The Fourth People's Hospital of Ningxia Hui Autonomous Region, Yinchuan 750021, China

Received:2024-09-19 Online:2025-07-23 Published:2025-07-25
Contact: DENG Guangcun E-mail:3414153939@qq.com;dgc@nxu.edu.cn

摘要/Abstract

摘要：

本研究旨在探究脂肪酸结合蛋白4(fatty acid-binding protein 4, FABP4)对卡介苗(Bacillus Calmette-Guérin, BCG)感染的巨噬细胞趋化因子分泌的调控作用。首先，本研究以小鼠巨噬细胞RAW264.7为靶细胞，利用小干扰RNA技术敲减FABP4的表达，并结合BCG感染，通过荧光定量PCR、ELISA等方法检测趋化因子的表达差异。随后以C57BL/6J小鼠为研究对象，先通过腹腔注射FABP4抑制剂BMS-309403抑制FABP4功能，随后以气道灌注的方式感染BCG，利用Western blot、ELISA检测肺脏趋化因子分泌水平；HE染色观察小鼠肺组织损伤；涂布平板法检测小鼠肺组织菌载量。结果显示：与未感染组相比，BCG感染的RAW264.7细胞趋化因子CCL2、CCL3、CXCL2、CXCL10、CXCL11的mRNA表达呈时间和浓度依赖性上调。并在感染复数为10感染时间为12 h时已显著(P < 0.05)高于对照组；与BCG组相比，FABP4敲减结合BCG组，细胞内与细胞上清中CCL3 (P < 0.001)、CXCL2 (P < 0.01)的表达量极显著下调，而CCL2、CXCL10、CXCL11的表达量无差异；小鼠研究发现，BMS-309403会降低小鼠肺组织以及血清中趋化因子CCL3 (P < 0.001)、CXCL2 (P < 0.001)的表达，并减弱BCG感染后C57BL/6J小鼠肺组织损伤。平板涂布法检测发现，BMS-309403极显著上调BCG感染后小鼠肺部菌的含量(P < 0.001)。综上表明，FABP4促进BCG感染的RAW264.7巨噬细胞、小鼠肺组织趋化因子的表达和肺部病菌的清除。

关键词: 卡介苗, 巨噬细胞, 脂肪酸结合蛋白4, 趋化因子

Abstract:

The aim of this study was to explore the regulatory role of fatty acid-binding protein 4 (FABP4) on chemokine secretion in Bacillus Calmette-Guérin (BCG) infected macrophages. Using mouse macrophage RAW264.7 as in vitro model, the expression of FABP4 was knocked down by siRNA, combined with BCG infection, and the expression of chemokines were detected by fluorescence quantitative PCR, ELISA and other methods. Subsequently, the function of FABP4 was inhibited by FABP4 inhibitor BMS-309403 in C57BL/6J mice, combined with BCG infection. The levels of lung chemokine secretion were detected by Western blot and ELISA. The lung tissue injury was observed by HE staining. The bacterial load in lung tissue of mice was detected by coated plate method. The results showed that the mRNA expressions of chemokines CCL2, CCL3, CXCL2, CXCL10 and CXCL11 of RAW264.7 macrophages infected with BCG were up-regulated in a time and dose dependent manner compared with the control group. The expression was significantly increased (P < 0.05) when the multiple of infection was 10 MOI at 12 h post infection. Compared with the BCG group, the expression levels of CCL3 (P < 0.001) and CXCL2 (P < 0.01) were significantly down-regulated in FABP4 knockdown combined with the BCG group, while the expression levels of CCL2, CXCL10 and CXCL11 were not significantly different. Studies in mice showed that BMS-309403 reduced the expression of chemokines CCL3 (P < 0.001) and CXCL2 (P < 0.001) in lung tissue and serum of mice, and mitigate the lung tissue injury of C57BL/6J mice after BCG infection. Plate coating found that BMS-309403 significantly increased the content of pulmonary bacteria in mice infected with BCG (P < 0.001). In conclusion, FABP4 promoted the expression of chemokines in RAW264.7 macrophages and mouse lung tissue infected with BCG and clearance of lung bacteria.

Key words: Bacillus Calmette-Guérin, macrophage, fatty acid-binding protein 4, chemokine

中图分类号:

S855.1

赵燕萍, 王晓平, 于嘉霖, 宫照乾, 吴晓玲, 邓光存. 脂肪酸结合蛋白4对卡介苗感染巨噬细胞趋化因子分泌的调控作用[J]. 畜牧兽医学报, 2025, 56(7): 3484-3494.

ZHAO Yanping, WANG Xiaoping, YU Jialin, GONG Zhaoqian, WU Xiaoling, DENG Guangcun. Role of Fatty Acid-Binding Protein 4 on Chemokine Secretion in Bacillus Calmette-Guérin Infected Macrophages[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3484-3494.

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引物名称 Primer name	引物序列(5′→3′) Sequence
CCL2	F: GCCTGCTGTTCACAGTTGC
CCL2	R: GAGTGGGGCGTTAACTGCAT
CCL3	F: GATTCCACGCCAATTCATCG
CCL3	R: TTCAGTTCCAGGTCAGTGATG
CXCL2	F: AATGCCTGAAGACCCTGC
CXCL2	R: TTTTGACCGCCCTTGAGAG
CXCL10	F: TTGCCCTTGGTCTTCTGA
CXCL10	R: GTCGCACCTCCACATAGC
CXCL11	F: CGGCTGCGACAAAGTTGAAG
CXCL11	R: TATGAGGCGAGCTTGCTTGG
β-Actin	F: GTGCTATGTTGCTCTAGACTTCG
β-Actin	R: ATGCCACAGGATTCCATACC

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脂肪酸结合蛋白4对卡介苗感染巨噬细胞趋化因子分泌的调控作用

Role of Fatty Acid-Binding Protein 4 on Chemokine Secretion in Bacillus Calmette-Guérin Infected Macrophages

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