miR-155-5P regulates interleukin-6 and macrophage inflammatory protein-2 expression in alveolar macrophages to influence neutrophil migration during acute lung injury in sepsis

doi:10.3877/cma.j.issn.2096-1537.2020.02.019

Abstract

Abstract:

Objective

To determine the regulatory effect of miR-155-5P on the expression of interleukin-6 (IL-6) and macrophage inflammatory protein-2 (MIP-2) in alveolar macrophages and the influence on neutrophil migration during acute lung injury (ALI) in sepsis.

Methods

Mouse alveolar macrophages (MH-S) were cultured and divided into a control group, sepsis group, and miR-155-5P inhibitor group. The miR-155-5P inhibitor group was transfected with miR-155-5P antagomir in advance. Lipopolysaccharide (LPS) was used to intervene the sepsis group and miR-155-5P inhibitor group for 12 h. RT-PCR was used to detect the expression of miR-155-5P, IL-6 mRNA, and MIP-2 mRNA in alveolar macrophages of the three groups, and ELISA was used to detect the concentrations of IL-6 and MIP-2 in cell suspension. Again, MH-S was divided into three groups and cultured in the lower compartment of transwell plate. Neutrophils labeled with CFSE were cultured in the upper compartment. After LPS intervention, fluorescence values of cells in the upper compartment and lower compartment were detected by flow cytometry, respectively, to calculate the migration rate of neutrophils.

Results

The expression of miR-155-5P (8.04±0.65), IL-6 mRNA (57.05±6.88), and MIP-2 mRNA (52.98±2.58) in MH-S cells in the sepsis group were significantly up-regulated compared with the control group (1.00±0.00, 1.00±0.00, and 1.00±0.00, respectively; P<0.05). The concentrations of IL-6 [(50.85±0.12) pg/ml] and MIP-2 [(69.96±1.40) pg/ml] and neutrophil migration rate (64.36%) were significantly higher sepsis group than in the control group [(38.58±0.13) pg/ml, (56.00±0.29) pg/ml, and 30.98%, respectively; P<0.05]. The expression of miR-155-5P in the miR-155-5P inhibitor group (0.19±0.35) was lower than that in the control group (P<0.05). The expression of IL-6 mRNA (39.66±3.65) and MIP-2 mRNA (31.01±2.88), and the concentrations of IL-6 [(42.45±1.08) pg/ml] and MIP-2 [(59.01±1.63) pg/ml] in the miR-155-5P inhibitor group were significantly higher than those of the control group (P<0.05), but lower than those of the sepsis group (P<0.05).

Conclusion

In ALI secondary to sepsis, miR-155-5P is highly expressed in alveolar macrophages, which is positively correlated with the expression of pro-inflammatory factors IL-6 and MIP-2. miR-155-5P promotes the production of IL-6 and MIP-2, and causes a large number of neutrophils to migrate into the lung, thus causing lung injury. Inhibiting the expression of miR-155-5P can inhibit the production of IL-6 and MIP-2, thus reducing the chemotactic migration of neutrophils and protecting the lung.

Key words: miR-155-5P, Interleukin-6, Macrophage inflammatory proteins, Macrophages, Neutrophils, Acute lung injury

Kai Liu, Lingling Jia, Qian Liu, Jie Sun, Xiwei Zhao, Wenkai Zhang. miR-155-5P regulates interleukin-6 and macrophage inflammatory protein-2 expression in alveolar macrophages to influence neutrophil migration during acute lung injury in sepsis[J]. Chinese Journal of Critical Care & Intensive Care Medicine(Electronic Edition), 2020, 06(02): 198-205.

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URL: https://icu.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2096-1537.2020.02.019

https://icu.cma-cmc.com.cn/EN/Y2020/V06/I02/198

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References 18

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组别	miR-155-5P	白介素6 mRNA	MIP-2 mRNA
对照组	1.00±0.00	1.00±0.00	1.00±0.00
脓毒症组	8.04±0.65	57.05±6.88	52.98±2.58
MiR-155-5P抑制物组	0.19±0.35	39.66±3.65	31.01±2.88
F值	398.61	122.13	409.17
P值	<0.001	<0.001	<0.001

组别	miR-155-5P	白介素6 mRNA	MIP-2 mRNA
对照组	1.00±0.00	1.00±0.00	1.00±0.00
脓毒症组	8.04±0.65	57.05±6.88	52.98±2.58
MiR-155-5P抑制物组	0.19±0.35	39.66±3.65	31.01±2.88
F值	398.61	122.13	409.17
P值	<0.001	<0.001	<0.001

组别	白介素6	MIP-2
对照组	38.58±0.13	56.00±0.29
脓毒症组	50.85±0.12	69.96±1.40
MiR-155-5P抑制物组	42.45±1.08	59.01±1.63
F值	295.58	103.03
P值	<0.001	<0.001

组别	白介素6	MIP-2
对照组	38.58±0.13	56.00±0.29
脓毒症组	50.85±0.12	69.96±1.40
MiR-155-5P抑制物组	42.45±1.08	59.01±1.63
F值	295.58	103.03
P值	<0.001	<0.001

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