鸢尾素对感染大肠埃希菌小鼠组织损伤及细菌清除的影响

doi:10.11843/j.issn.0366-6964.2024.11.042

摘要/Abstract

摘要：

本研究旨在探究鸢尾素(irisin)对大肠埃希菌(Escherichia coli, E. coli)感染小鼠的组织损伤及巨噬细胞清除细菌和诱发炎症的影响。按半数致死剂量经腹腔感染E. coli建立小鼠模型，通过体温、临床评分、生存率、器官组织损伤程度、组织中细菌载量和炎症水平对鸢尾素的保护效果进行评估；在体外将不同浓度鸢尾素与E. coli孵育，利用酶标仪检测菌液OD_{600 nm}，评价鸢尾素是否具有抑菌效果；鸢尾素预处理小鼠单核巨噬细胞系(RAW264.7)后，使用E. coli感染细胞，收集细胞裂解液和mRNA评价巨噬细胞吞噬和清除细菌的能力以及M1极化水平。结果表明，鸢尾素能够明显提高细菌感染小鼠的生存率，降低临床评分，促进体温恢复，减轻肝、肺和肾的损伤(P < 0.000 1)，降低心(P < 0.05)、肝(P < 0.01)、脾(P < 0.01)、肺(P < 0.01)和肾(P>0.05)的细菌载量，降低肝脏炎症因子白细胞介素(interleukin，IL)-1β(P < 0.000 1)和IL-6(P < 0.001)mRNA水平及肾脏炎症因子IL-1β(P < 0.05)和IL-6(P < 0.01) mRNA水平；在体外鸢尾素对E. coli不具有抑菌效果，但可以促进RAW 264.7吞噬(P < 0.001)和清除E. coli，还能降低RAW 264.7的炎症标志物IL-1β(P < 0.000 1)、IL-6(P < 0.000 1)和iNOS(P < 0.001)mRNA水平。鸢尾素能够促进巨噬细胞吞噬和清除细菌，降低细菌感染诱导的巨噬细胞促炎反应，减轻细菌感染小鼠的组织损伤。

关键词: 鸢尾素, 巨噬细胞, 炎症, 细菌感染

Abstract:

This study was conducted to investigate the effects of irisin on Escherichia coli (E. coli)-induced tissue damage in mice, and bacterial clearance and inflammation induced by macrophages. The mouse model was established by intraperitoneal infection with E. coli at a median lethal dose. The protective effect of irisin was evaluated by body temperature, clinical score, survival rate, degree of organ and tissue damage, bacterial load and inflammation level of tissue. E. coli was incubated with different concentrations of irisin in vitro, and the bacterial solution OD_{600 nm} was detected by microplate reader to evaluate the antibacterial effect of irisin. After pretreatment of mouse mononuclear macrophages cells (RAW264.7) with irisin, macrophages were infected with E. coli, and cell lysates and mRNA were collected to evaluate the ability of macrophages to engulf and clear bacteria and the level of M1 polarization. Results showed that irisin could significantly improve the survival rate of bacterially infected mice, reduce the clinical score, promote body temperature recovery, and reduce the liver, lung and kidney damage (P < 0.000 1), decreased the bacterial load of heart (P < 0.05), liver (P < 0.01), spleen (P < 0.01), lung (P < 0.01), kidney (P>0.05), and reduced IL-1β(P < 0.000 1) and IL-6 (P < 0.001) mRNA levels in liver and interleukin (IL)-1β (P < 0.05) and IL-6 (P < 0.01) mRNA levels in kidney. Irisin did not inhibit the growth of E. coli in vitro, but promoted RAW264.7 phagocytosis (P < 0.001) and clearance of E. coli, and also reduced the inflammatory marker IL-1β(P < 0.000 1), IL-6 (P < 0.000 1) and iNOS (P < 0.001) mRNA levels. Irisin can promote the phagocytosis and clearance of bacteria by macrophages, reduce the pro-inflammatory response of macrophages induced by bacterial infection, and alleviate tissue damage in mice.

Key words: irisin, macrophage, inflammation, bacterial infection

中图分类号:

安一娜, 谭姝瑜, 冯岚迪, 鲍明悦, 刘梦晗, 尹勤昊, 董彦君. 鸢尾素对感染大肠埃希菌小鼠组织损伤及细菌清除的影响[J]. 畜牧兽医学报, 2024, 55(11): 5267-5277.

Yina AN, Shuyu TAN, Landi FENG, Mingyue BAO, Menghan LIU, Qinhao YIN, Yanjun DONG. Effects of Irisin on Tissue Damage and Bacterial Clearance in Mice Infected with Escherichia coli[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5267-5277.

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观测指标 Index of observation	评分标准 Standard for evaluation	评分 Score
外貌 Appearance	正常	0
外貌 Appearance	毛杂乱	1
	立毛	2
	耸起	3
	具有上述的现象并且眼睛半闭	4
行为举止(未刺激) Behavior and demeanor (unstimulated)	正常	0
行为举止(未刺激) Behavior and demeanor (unstimulated)	轻微的改变	1
	较少移动并且聚集在一起	2
	表现出不安或非常安静	3
行为举止(刺激) Behavior and demeanor (stimulated)	作出反应并警觉	0
行为举止(刺激) Behavior and demeanor (stimulated)	反应迟钝并且不警觉	3
临床体征 Clinical signs	正常的呼吸速率	0
临床体征 Clinical signs	呼吸轻微的改变	1
	腹部呼吸速率下降	2
	明显的腹部呼吸和发绀	3
水合状态 Hydration status	正常	0
水合状态 Hydration status	脱水	5

基因名称 Gene name	引物序列(5′→3′) Primer sequence
UCP1	F-ATGTTTGTGTGCTTTGAA
	R-CAGTATCTCTTCCTCCAA
IL-6	F-GGGAAATCGTGGAAATGAGA
	R-TTCTGCAAGTGCATCATCGT
IL-1β	F-CAACCAACAAGTGATATTCTCCATG
	R-GATCCACACTCTCCAGCTGCA
iNOS	F-TTTGGGAATGGAGACTGT
	R-AAGGTGTGGTTGAGTTCT
GAPDH	F-ACCACCATGGAGAAGGCCGG
	R- CTCAGTGTAGCCCAAGATGC

鸢尾素浓度/(μg·mL^-1) Irisin concentration	菌液OD_{600 nm}值 OD_{600 nm} of bacterial solution
0.000	0.646 35
0.125	0.667 50
0.250	0.631 50
0.500	0.627 90
1.000	0.636 45
2.000	0.645 75
4.000	0.642 90
8.000	0.634 30
16.000	0.653 80
32.000	0.665 50

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