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circHECTD1 attenuates apoptosis of alveolar epithelial cells in acute lung injury

Abstract

Circular RNAs (circRNAs) play important roles in many lung diseases. This study aimed to investigate the role of circHECTD1 in acute lung injury (ALI). The mouse and cell models of ALI were induced by lipopolysaccharide (LPS). The apoptosis of alveolar epithelial cells (AECs) was detected by flow cytometry. The relationships between circHECTD1, miRNAs, and target genes were assessed by RNA pull-down, luciferase reporter gene, and RNA-FISH assays. circHECTD1 was downregulated in LPS-induced human and mouse AECs (HBE and MLE-12). The knockdown of circHECTD1 increased the apoptotic rates and the expressions of miR-136 and miR-320a, while its overexpression caused opposite effects in LPS-induced HBE and MLE-12 cells. Mechanistically, circHECTD1 bound to miR-320a and miR-136. miR-320a targeted PIK3CA and mediated the effect of circHECTD1 on PIK3CA expression. miR-136 targeted Sirt1 and mediated the effect of circHECTD1 on Sirt1 expression. Silencing PIK3CA and/or Sirt1 reversed the effect of circHECTD1 overexpression on the apoptosis of LPS-induced HBE and MLE-12 cells. In vivo, overexpression of circHECTD1 alleviated the LPS-induced ALI of mice. Our findings suggested that circHECTD1 inhibits the apoptosis of AECs through miR-320a/PIK3CA and miR-136/Sirt1 pathways in LPS-induced ALI.

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Fig. 1: The expression of circHECTD1 in LPS-induced alveolar epithelial cells (AECs).
Fig. 2: The effect of circHECTD1 on the apoptosis of AECs.
Fig. 3: circHECTD1 binds to miR-136 and miR-320 in AECs.
Fig. 4: circHECTD1 regulates PIK3CA expression through miR-320a.
Fig. 5: circHECTD1 regulates Sirt1 expression through miR-136.
Fig. 6: Sirt1 and PIK3CA mediate the effect of circHECTD1 on apoptosis of AECs.
Fig. 7: circHECTD1 relieves ALI of mice. ALI mice were divided into two groups: the Ad-circHECTD1 group (n = 7) and the Ad-GFP group (n = 7).

Data availability

All the data generated or analyzed during this study are included in the manuscript.

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Funding

This study was supported by the finding of the Key scientific research projects of henan provincial colleges and universities (NO. 19A320009).

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L.H.B. and G.M. participated in the experimental design, manuscript writing and manuscript revision. L.H.B., N.X.X., S.H.J., F.M., D.Y.M., S.R.Q., and M.N.L. participated in cell experiments, acquisition of data, data analysis and interpretation. W.H.L. and W.D. participated in animal experiment and histological experiment. All authors read, revised, and approved the final manuscript.

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Correspondence to Hongbin Li or Min Gao.

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The authors declare no competing interests.

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The animal experiments were performed in the Laboratory Animal Center of Zhengzhou University and approved by the Animal Ethics Committee of the First Affiliated Hospital of Zhengzhou University.

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Li, H., Niu, X., Shi, H. et al. circHECTD1 attenuates apoptosis of alveolar epithelial cells in acute lung injury. Lab Invest 102, 945–956 (2022). https://doi.org/10.1038/s41374-022-00781-z

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  • DOI: https://doi.org/10.1038/s41374-022-00781-z

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