Angiotensin-(1-7) attenuates SARS-CoV2 spike protein-induced interleukin-6 and interleukin-8 production in alveolar epithelial cells through activation of Mas receptor

Yi-Luen Shen; Yi-An Hsieh; Po-Wei Hu; Po-Chun Lo; Yi-Han Hsiao; Hsin-Kuo Ko; Fang-Chi Lin; Chien-Wen Huang; Kang-Cheng Su; Diahn-Warng Perng

doi:10.1016/j.jmii.2023.09.003

Angiotensin-(1-7) attenuates SARS-CoV2 spike protein-induced interleukin-6 and interleukin-8 production in alveolar epithelial cells through activation of Mas receptor

J Microbiol Immunol Infect. 2023 Dec;56(6):1147-1157. doi: 10.1016/j.jmii.2023.09.003. Epub 2023 Sep 22.

Authors

Yi-Luen Shen¹, Yi-An Hsieh¹, Po-Wei Hu², Po-Chun Lo³, Yi-Han Hsiao⁴, Hsin-Kuo Ko⁴, Fang-Chi Lin⁴, Chien-Wen Huang⁵, Kang-Cheng Su⁶, Diahn-Warng Perng⁷

Affiliations

¹ Division of Chest Medicine, Department of Internal Medicine, Asia University Hospital, Taichung, Taiwan, ROC.
² Division of Chest Medicine, Department of Internal Medicine, National Yang Ming Chiao Tung University Hospital, Taiwan, ROC; School of Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC.
³ Taipei Veterans General Hospital, Fenglin Branch, Hualien, Taiwan, ROC; Department of Chest Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, ROC.
⁴ School of Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC; Department of Chest Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, ROC.
⁵ Division of Chest Medicine, Department of Internal Medicine, Asia University Hospital, Taichung, Taiwan, ROC; Department of Medical Laboratory Science and Biotechnology, College of Medical and Health Science, Asia University, Taichung, Taiwan, ROC.
⁶ School of Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC; Department of Chest Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, ROC. Electronic address: kcsu@vghtpe.gov.tw.
⁷ School of Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan, ROC; Department of Chest Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, ROC. Electronic address: dwperng@vghtpe.gov.tw.

PMID: 37802686
DOI: 10.1016/j.jmii.2023.09.003

Abstract

Background: SARS-CoV-2 spike proteins (SP) can bind to the human angiotensin-converting enzyme 2 (ACE2) in human pulmonary alveolar epithelial cells (HPAEpiC) and trigger an inflammatory process. Angiotensin-(1-7) may have an anti-inflammatory effect through activation of Mas receptor. This study aims to investigate whether SARS-CoV-2 SP can induce inflammation through ACE2 in the alveolar epithelial cells which can be modulated through angiotensin-(1-7)/Mas receptor axis.

Methods: HPAEpiC were treated with SARS-CoV-2 SP in the presence or absence of ACE2 antagonist-dalbavancin and Mas receptor agonist-angiotensin-(1-7). Proinflammatory cytokine production (IL-6 and IL-8) were measured at mRNA and protein levels. MAP kinase phosphorylation and transcription factor activation was determined by Western Blot. Mas receptor was blocked by either antagonist (A779) or knockdown (specific SiRNA). Experiments were replicated using A549 cells.

Findings: SARS-CoV-2 SP (5 μg/mL) significantly induced MAP kinase (ERK1/2) phosphorylation, downstream transcription factor (activator protein-1, AP-1) activation and cytokine production (IL-6 and IL-8) at both mRNA and protein levels. Pretreatment with dalbavancin (10 μg/mL), or angiotensin-(1-7) (10 μM) significantly reduced ERK1/2 phosphorylation, AP-1 activation, and cytokine production. However, these angiotensin-(1-7)-related protective effects were significantly abolished by blocking Mas receptor with either antagonist (A799,10 μM) or SiRNA knockdown.

Interpretation: SARS-CoV-2 SP can induce proinflammatory cytokine production, which can be inhibited by either ACE2 antagonist or Mas receptor agonist-angiotensin-(1-7). Angiotensin-(1-7)-related protective effect on cytokine reduction can be abolished by blocking Mas receptor. Our findings suggest that ACE2/angiotensin-(1-7)/Mas axis may serve as a therapeutic target to control inflammatory response triggered by SARS-CoV-2 SP.

Keywords: ACE; Angiotensin-(1–7); COVID-19; SARS-CoV-2; Spike protein.

MeSH terms

Alveolar Epithelial Cells / metabolism
Angiotensin-Converting Enzyme 2
COVID-19*
Cytokines
Humans
Interleukin-6* / metabolism
Interleukin-8
Peptidyl-Dipeptidase A / metabolism
RNA, Messenger
RNA, Small Interfering / metabolism
RNA, Viral
SARS-CoV-2 / genetics
Spike Glycoprotein, Coronavirus
Transcription Factor AP-1

Substances

angiotensin I (1-7)
Angiotensin-Converting Enzyme 2
Cytokines
Interleukin-6
Interleukin-8
Peptidyl-Dipeptidase A
RNA, Messenger
RNA, Small Interfering
RNA, Viral
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2
Transcription Factor AP-1
IL6 protein, human
CXCL8 protein, human