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miR-155 activates the NLRP3 inflammasome by regulating the MEK/ERK/NF-κB pathway in carotid atherosclerotic plaques in ApoE−/− mice

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Abstract

Atherosclerosis is an inflammatory disease. The NLRP3 inflammasome and miR-155 are significant components of inflammation and atherosclerosis. The aim of this research was to explore the possible mechanism by which miR-155 mediates the formation of carotid atherosclerotic plaques via the NLRP3 inflammasome. Fifty 6-week-old male ApoE−/− mice were randomly divided into 5 groups. They are the blank group, the negative control (NC) group, the miR-155 mimic group, the miR-155 inhibitor group, and the miR-155 mimic and ERK inhibitor group. The blood lipid levels were measured by the enzyme method Oil red O, HE, and immunohistochemical staining were used to observe the degree of carotid plaque formation. PCR was used to measure the expression of miR-155. The blood lipid levels were measured by the enzyme method. Western blotting was used to measure the expression of NLRP3 inflammasome-related proteins, interleukin-1β, interleukin-18, and MEK/ERK/NF-κB signaling pathway-related proteins. Compared with those of the NC group, the expression of miR-155 in the miR-155 mimic group increased significantly (P < 0.05), the degree of carotid plaque formation increased, the plasma levels of TC and LDL also increased significantly (P < 0.05); the expression levels of NLRP3 inflammasome-related proteins, interleukin-1β, interleukin-18, and MEK/ERK/NF-κB signaling pathway-related proteins were also significantly increased. Injection of ERK inhibitors into miR-155 mimic mice reduced the expression levels of p-NF-κB, NLRP3 inflammasome-related proteins, and inflammatory cytokines. In conclusion, miR-155 can promote the formation of atherosclerotic plaque in ApoE−/− mice, which may be achieved by regulating the MEK/ERK/NF-κB pathway to activate the NLRP3 inflammasome.

Highlights

• In ApoE−/− mice, miR-155 promotes atherosclerotic plaque formation.

• The NLRP3 inflammasome has an important role in the inflammatory process of atherosclerosis.

• miR-155 activates the NLRP3 inflammasome by regulating the MEK/ERK/NF-κB pathway in carotid atherosclerotic plaques of ApoE−/− mice.

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Abbreviations

BCA:

Bicinchoninic acid

DAMPs:

Danger-associated molecular patterns

HDL:

High-density lipoprotein

HE:

Hematoxylin-eosin

IHC:

Immunohistochemical

LDL:

Low-density lipoprotein cholesterol

MAPK:

Mitogen-activated protein kinase

MiRNAs:

MicroRNAs

NC:

Negative control

NF-κB:

Nuclear factor κ-light-chain-enhancer of activated B cells

NLR:

NOD-like receptor

NLRP3:

NOD-like receptor family pyrin domain containing 3

PAMPs:

Pathogen-associated molecular patterns

P-IL-1β:

Pro-interleukin-1β

P-IL-18:

Pro-interleukin-18

PRR:

Pattern recognition receptor

RA:

Rheumatoid arthritis

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SnRNA:

Small nuclear RNA

TC:

Total cholesterol

TG:

Triglyceride

VSMCs:

Vascular smooth muscle cells

GvHD:

Graft-versus-host disease

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Acknowledgements

This project is based on the item subsidized by the National Key R&D Program of China (No. 2017YFC1310903) and the National Natural Sciences Foundation of China (No. 81771259 and No.81971111).

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Author notes

  1. Qing Peng and Ruihua Yin contributed equally to this work.

Authors and Affiliations

  1. Department of Neurology, The Affiliated Hospital of Qingdao University, Qingdao, 266003, Shandong, People’s Republic of China

    Jing Wang & Aijun Ma

  2. Department of Critical Care Medicine, The Affiliated Hospital of Qingdao University, Qingdao, 266033, People’s Republic of China

    Xiaoyan Zhu

  3. Institute of Cerebrovascular Diseases, The Affiliated Hospital of Qingdao University, Qingdao, 266003, People’s Republic of China

    Liying Jin

  4. Department of Neurology and Institute of Cerebrovascular Diseases, The Affiliated Hospital of Qingdao University, Qingdao, 266003, Shandong, People’s Republic of China

    Xudong Pan & Aijun Ma

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  1. Qing Peng
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Contributions

AM: conceptualization and supervision. XP: methodology and conceptualization. QP: investigation, methodology, and writing-original draft. RY: investigation and formal analysis. XZ: writing-reviewing and editing. LJ: formal analysis and resources. JW: resources. The authors declare that all data were generated in-house and that no paper mill was used.

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Correspondence to Xudong Pan or Aijun Ma.

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Peng, Q., Yin, R., Zhu, X. et al. miR-155 activates the NLRP3 inflammasome by regulating the MEK/ERK/NF-κB pathway in carotid atherosclerotic plaques in ApoE−/− mice. J Physiol Biochem 78, 365–375 (2022). https://doi.org/10.1007/s13105-022-00871-y

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