重组融合蛋白IL-18对金黄色葡萄球菌感染小鼠免疫相关炎症因子表达的影响

陈晨, 陈强, 李岚, 余晓君, 柯江维, 何美娟, 周红平, 杨文萍, 王文杏

中国当代儿科杂志 ›› 2017, Vol. 19 ›› Issue (6) : 705-711.

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中国当代儿科杂志 ›› 2017, Vol. 19 ›› Issue (6) : 705-711. DOI: 10.7499/j.issn.1008-8830.2017.06.018
论著·实验研究

重组融合蛋白IL-18对金黄色葡萄球菌感染小鼠免疫相关炎症因子表达的影响

  • 陈晨, 陈强, 李岚, 余晓君, 柯江维, 何美娟, 周红平, 杨文萍, 王文杏
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Effects of recombinant fusion protein interleukin-18 on expression of immune-inflammatory factors in mice infected with Staphylococcus aureus

  • CHEN Chen, CHEN Qiang, LI Lan, YU Xiao-Jun, KE Jiang-Wei, HE Mei-Juan, ZHOU Hong-Ping, YANG Wen-Ping, WANG Wen-Xing
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摘要

目的 观察重组融合蛋白IL-18对金黄色葡萄球菌(SA)感染后小鼠体内免疫相关炎症因子表达的影响,探讨IL-18在体内防御SA感染的可能机制。方法 将40只SPF级雌性BLAB/c小鼠随机分为对照组、SA感染组、免疫组和干预组。采用鼻腔接种SA液建立SA感染小鼠模型,免疫组和干预组均在建模前以IL-18滴鼻,但免疫组不予SA接种,对照组以PBS进行替代处理。采用ELISA法测定各组小鼠血液及支气管肺泡灌洗液(BALF)中IL-4、干扰素-γ(IFN-γ)、肿瘤坏死因子(TNF)、粒细胞集落刺激因子(G-CSF)、IgM的浓度。实时荧光定量PCR技术检测各组小鼠肺组织中巨噬细胞炎性蛋白(MIP)-1α和MIP-2β mRNA表达水平。结果 与对照组相比,SA感染组和免疫组小鼠血清及BALF中IL-4、G-CSF、IgM浓度,以及肺组织中MIP-1α、MIP-2β mRNA含量均升高(P < 0.05);SA感染组小鼠血清及BALF中IFN-γ水平降低,TNF水平升高(P < 0.05);免疫组小鼠血清及BALF中IFN-γ水平升高(P < 0.05)。与SA感染组相比,干预组小鼠血清及BALF中IL-4、IFN-γ、G-CSF、IgM浓度,以及肺组织中MIP-1α mRNA含量均升高,血清及BALF中TNF水平,以及肺组织中MIP-2β mRNA含量均降低(P < 0.05)。除血清IFN-γ水平外,其余上述指标在干预组小鼠中均高于对照组(P < 0.05)。结论 重组融合蛋白IL-18经黏膜免疫小鼠,可改变SA感染后小鼠血清及BALF中炎症因子,以及肺组织中MIP-1α、MIP-2β mRNA的表达水平,从而促进机体的抗感染免疫反应,增强了机体清除病原体的能力。

Abstract

Objective To observe the effects of recombinant fusion protein interleukin (IL)-18 on the expression of immune-inflammatory factors in the mice infected with Staphylococcus aureus (SA), and to investigate the mechanism of action of IL-18 in defense of SA infection in vivo. Methods A total of 40 specific pathogen-free female BLAB/c mice were randomly divided into four groups:control, SA infection, immunized, and intervention. A mouse model of SA infection was established by nasal inoculation with SA liquid. The immunized group and the intervention group were intranasally given IL-18 before SA modeling, and then the SA infection group and the intervention group received the nasal inoculation with SA liquid; the control group was treated with phosphate buffered saline instead. The levels of IL-4, interferon (IFN)-γ, tumor necrosis factor (TNF), granulocyte colony-stimulating factor (G-CSF), IgM in the serum and bronchoalveolar lavage fluid (BALF) of mice were measured by enzyme-linked immunosorbent assay. The expression of macrophage inflammatory protein (MIP)-1α mRNA and MIP-2β mRNA in the lung tissue of mice were determined by real-time fluorescent quantitative PCR. Results Compared with the control group, the SA infection group and the immunized group had significantly higher levels of IL-4, G-CSF, and IgM in the serum and BALF and expression of MIP-1α mRNA and MIP-2β mRNA in the lung tissue (P < 0.05); the SA infection group had a significantly lower level of IFN-γ and a significantly higher level of TNF in the serum and BALF (P < 0.05); the immunized group had a significantly higher level of IFN-γ in the serum and BALF (P < 0.05). Compared with the SA infection group, the intervention group had significantly higher levels of IL-4, IFN-γ, G-CSF, and IgM in the serum and BALF and expression of MIP-1α mRNA in the lung tissue. In contrast, the intervention group showed a significantly lower level of TNF in the serum and BALF and expression of MIP-2β mRNA in the lung tissue (P < 0.05). All the above indicators in the intervention group were significantly higher than those in the control group (P < 0.05), except the serum level of IFN-γ. Conclusions In the mice infected with SA, the recombinant fusion protein IL-18 by mucosal immunity can affect inflammatory factors in the serum and BALF and the expression of MIP-1α mRNA and MIP-2β mRNA in the lung tissue to promote the anti-infective immune response and enhance the ability to clear pathogens.

关键词

IL-18 / 金黄色葡萄球菌 / 炎症因子 / 巨噬细胞炎性蛋白 / 小鼠

Key words

IL-18 / Staphylococcus aureus / Inflammatory factor / Macrophage inflammatory protein / Mice

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陈晨, 陈强, 李岚, 余晓君, 柯江维, 何美娟, 周红平, 杨文萍, 王文杏. 重组融合蛋白IL-18对金黄色葡萄球菌感染小鼠免疫相关炎症因子表达的影响[J]. 中国当代儿科杂志. 2017, 19(6): 705-711 https://doi.org/10.7499/j.issn.1008-8830.2017.06.018
CHEN Chen, CHEN Qiang, LI Lan, YU Xiao-Jun, KE Jiang-Wei, HE Mei-Juan, ZHOU Hong-Ping, YANG Wen-Ping, WANG Wen-Xing. Effects of recombinant fusion protein interleukin-18 on expression of immune-inflammatory factors in mice infected with Staphylococcus aureus[J]. Chinese Journal of Contemporary Pediatrics. 2017, 19(6): 705-711 https://doi.org/10.7499/j.issn.1008-8830.2017.06.018

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