肠道菌群对高脂饮食诱导肥胖小鼠糖脂代谢紊乱的免疫调节作用

doi:10.3969/j.issn.1006-5725.2024.11.007

摘要/Abstract

摘要：

目的探讨混合抗生素作用于小鼠肠道菌群从而影响机体免疫调节，探索肠道菌群在肥胖发生发展中的作用，为肥胖防治提供新思路与途径。方法 72只10周龄C57BL/6雄性小鼠随机分为空白对照（Ctrl）组、高脂喂养（HF）组、抗生素（ABX）组和联合（COMB）组（n = 18）。前2周（灌胃干预周）Ctrl组和HF组采用生理盐水灌胃，ABX组和COMB组采用混合抗生素灌胃，灌胃体积均为0.2 mL/（只·d）；后8周（饲料喂养周）Ctrl组和ABX组采用普通饲料喂养，HF组和COMB组采用高脂饲料喂养。每周测量小鼠体质量，灌胃干预周前后、饲料喂养第4、8周测量小鼠空腹血糖；实验结束时进行口服糖耐量实验；测定脏器系数，镜下观察白色和棕色脂肪组织细胞形态，测定血清游离脂肪酸、高密度脂蛋白、低密度脂蛋白、甘油三酯、总胆固醇含量，ELISA法检测血清TNF-α、IL-10、IL-4、IL-13、IL-33、MCP-1含量；小鼠粪便进行二代测序。结果高脂饮食引起小鼠体质量增加，血清总胆固醇、低密度脂蛋白、IL-13、IL-33、TNF-α、MCP-1含量增加，糖耐量和脏器系数降低（P < 0.05）。饲喂第1周至实验结束，COMB组体质量低于HF组（P < 0.05）。COMB组较HF组糖耐量、血清总胆固醇、低密度脂蛋白、IL-13、IL-33、TNF-α、MCP-1含量降低（P < 0.05）。HF组白色脂肪组织可见微血管充血出血，脂肪细胞不完整且极度膨胀，细胞质极度压缩；COMB组白色脂肪组织未见充血出血，脂肪细胞较完整。ABX组肠道菌群α多样性低于Ctrl组（P < 0.05）；ABX和HF组分别与Ctrl和COMB组微生物群落组成较为相近。结论高脂饮食诱导小鼠肥胖、糖脂代谢紊乱和机体炎症，短期混合抗生素使用能够调节小鼠肠道菌群，介导相关抗炎因子表达增加，上调宿主免疫，改善小鼠糖脂代谢。

关键词: 肠道菌群, 高脂饮食, 抗生素, 糖脂代谢, 免疫

Abstract:

Objective To explore the effect of mixed antibiotics on the intestinal flora of mice to affect the immune regulation of the body， explore the role of intestinal flora in the development of obesity， and provide new ideas and ways for the prevention and treatment of obesity. Methods Seventy-two 10-week-old male C57BL/6 mice were randomly divided into blank control （Ctrl） group， high-fat diet （HF） group， antibiotic （ABX） group， and combined （COMB） group （n = 18）. At the first 2 weeks （lavage intervention weeks）， Ctrl and HF group were given normal saline gavage； ABX and COMB group were given mixed antibiotics gavage， and the gavage volume was 0.2 mL/ animal/day. For the following 8 weeks （feeding weeks）， Ctrl and ABX group were fed with ordinary diet， HF and COMB group were fed with high-fat diet. Body weight was measured weekly， and fasting blood glucose was measured before and after gavage， and at the 4th and 8th week of feeding. Oral glucose tolerance test was performed at the end of the experiment. The organ coefficient was measured and the cell morphology of white and brown adipose tissue was observed. Serum was collected for the determination of free fatty acid， high-density lipoprotein， low-density lipoprotein， triglyceride， and total cholesterol. Serum TNF-α， IL-10， IL-4， IL-13， IL-33 and MCP-1 was detected by ELISA. The stool of mice was collected for second generation sequencing. Results High-fat diet increased body weight， serum total cholesterol， low-density lipoprotein， IL-13， IL-33， TNF-α， MCP-1 content， and decreased glucose tolerance and organ coefficient in mice （P < 0.05）. From the first feeding week to the end of the experiment， body weight in COMB group was significantly lower than that in HF group（P < 0.05）. The level of glucose tolerance， serum total cholesterol， low density lipoprotein， IL-13， IL-33， TNF-α and MCP-1 in COMB group was lower than those in HF group （P < 0.05）. The α diversity of intestinal flora in ABX group was lower than that in Ctrl group （P < 0.05）. Congestion and bleeding in WAT were obvious in HF group， but not in COMB group. The microbial community composition of ABX and HF group was similar to that of Ctrl and COMB group， respectively. Conclusion High-fat diet induces obesity， disorder of glucose and lipid metabolism and inflammation in mice. Short-term mixed antibiotic use can regulate the intestinal flora of mice， mediate increased expression of related anti-inflammatory factors， up-regulate host immunity， and improve glucose and lipid metabolism in mice.

Key words: intestinal flora, high-fat diet, antibiotic, glucose and lipid metabolism, immune

中图分类号:

R392.6

毕馨文,崔远捷,陆秋娴,崔佳,卜凡,何方,杨华,李鸣. 肠道菌群对高脂饮食诱导肥胖小鼠糖脂代谢紊乱的免疫调节作用[J]. 实用医学杂志, 2024, 40(11): 1505-1512.

Xinwen BI,Yuanjie CUI,Qiuxian LU,Jia CUI,Fan BU,Fang HE,Hua YANG,Ming. LI. Immunomodulatory effects of intestinal flora on glucose and lipid metabolism disorders in high⁃fat diet induced obese mice[J]. The Journal of Practical Medicine, 2024, 40(11): 1505-1512.

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物种	Ctrl组	HF组	ABX组	COMB组
厚壁菌门（Firmicutes）	50.66 ± 6.67	58.10 ± 6.36	45.70 ± 7.71	47.42 ± 5.99
拟杆菌门（Bacteroidota）	31.54 ± 7.22	19.00 ± 3.63	44.07 ± 8.41	23.50 ± 5.19^b
变形菌门（Proteobacteria）	4.12 ± 1.82	12.22 ± 3.97	1.96 ± 0.66	10.03 ± 2.70^b
疣微菌门（Verrucomicrobiota）	4.22 ± 1.70	1.70 ± 0.69	1.00 ± 0.31	7.37 ± 2.43^ab
脱铁杆菌门（Deferribacterota）	0.03 ± 0.02	0.08 ± 0.04	0.00 ± 0.00	0.03 ± 0.01^b
F/B值	4.46 ± 1.96	6.20 ± 1.50	2.71 ± 1.22	4.55 ± 1.33

物种	Ctrl组	HF组	ABX组	COMB组
另枝菌属（Alistipes）	2.52 ± 0.51	1.17 ± 0.30^a	2.40 ± 0.76	3.62 ± 1.20
粪杆菌属（Faecalibaculum）	1.27 ± 0.37	6.19 ± 2.10^a	2.35 ± 0.55	7.93 ± 2.50^ac
阿克曼菌属（Akkermansia）	4.21 ± 1.70	1.70 ± 0.69	1.00 ± 0.31	7.37 ± 2.43^bc
拟杆菌属（Bacteroides）	0.77 ± 0.21	3.21 ± 0.79^a	1.83 ± 0.43^a	6.83 ± 2.44^a
拟普雷沃氏菌属（Alloprevotella）	1.44 ± 0.52	1.12 ± 0.32	0.91 ± 0.29	1.19 ± 0.05^abc
嗜胆菌属（Bilophila）	0.03 ± 0.01	0.50 ± 0.31^a	0.08 ± 0.02	0.04 ± 0.03^b

分组	IL-10（pg/ mL）	IL-4（pg/ mL）	IL-13（pg/ mL）	IL-33（pg/ mL）	MCP-1（pg/ mL）	TNF-α（pg/ mL）
Ctrl组	268.18 ± 10.17	114.01 ± 3.77	22.11 ± 1.06	50.47 ± 3.58	7.56 ± 0.68	278.91 ± 15.00
HF组	178.02 ± 7.78^a	74.85 ± 4.08^a	36.48 ± 1.22^a	96.22 ± 3.66^a	14.70 ± 0.67^a	452.43 ± 16.00^a
ABX组	267.28 ± 7.26	106.23 ± 2.98^a	25.57 ± 1.03^a	56.72 ± 2.79^a	8.81 ± 0.48^a	292.01 ± 11.63^a
COMB组	238.56 ± 6.19^bc	95.23 ± 2.77^bc	28.06 ± 1.05^bc	67.21 ± 2.56^bc	11.06 ± 0.40^bc	346.82 ± 9.66^bc

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