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Intestinal epithelial Krüppel-like factor 4 alleviates endotoxemia and atherosclerosis through improving NF-κB/miR-34a-mediated intestinal permeability

Acta Pharmacologica Sinica (2024)Cite this article

Abstract

Maintenance of intestinal barrier function contributes to gastrointestinal homeostasis and therefore cardiovascular diseases. A number of studies show that intestinal permeability is affected by excessive inflammatory responses. Krüppel-like factor (KLF) 4 is one of the critical transcriptional factors, which controls multiple immune responses. In this study we investigated the role of KLF4 in regulating intestinal inflammation and permeability during the atherosclerotic process. Atherosclerotic model was established in ApoE−/− mice by feeding a high fat high cholesterol (HFHC) diet. We showed that colon expression levels of KLF4 and tight junction proteins were significantly decreased whereas inflammatory responses increased in atherosclerotic mice. Overexpression of colon epithelial Klf4 decreased atherosclerotic plaque formation and vascular inflammation in atherosclerotic mice, accompanied by remarkable suppression of intestinal NF-κB activation. We found that overexpression of epithelial Klf4 in atherosclerotic mice significantly increased intestinal tight junction expression and ameliorated endotoxemia, whereas replenishment of LPS abolished these benefits. Overexpression of Klf4 reversed LPS-induced permeability and downregulation of ZO-1 and Occludin in Caco-2 cells in vitro. HFHC diet stimulated the expression of epithelial microRNA-34a, whereas silence of epithelial Klf4 abolished the benefits of microRNA-34a sponge, a specific miR-34a inhibitor, on intestinal permeability and atherosclerotic development. A clinical cohort of 24 atherosclerotic patients supported colon KLF4/NF-κB/tight junction protein axis mediated intestine/cardiovascular interaction in patients with atherosclerosis. Taken together, intestinal epithelial KLF4 protects against intestinal inflammation and barrier dysfunction, ameliorating atherosclerotic plaque formation.

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Fig. 1: Intestinal expression of KLF4 is associated with intestinal inflammation and plaque formation in atherosclerotic mice.
Fig. 2: Intestinal overexpression of epithelial Klf4 alleviates atherosclerotic plaque formation and vascular inflammation in atherosclerotic mice.
Fig. 3: Intestinal overexpression of Klf4 decreases intestinal permeability in atherosclerotic mice.
Fig. 4: KLF4 inhibited NF-κB activation-mediated colic inflammation.
Fig. 5: Supplementation of lipopolysaccharide abolishes the anti-atherosclerotic benefits of intestinal overexpression of Klf4.
Fig. 6: Intestinal miR-34a/KLF4 signaling mediates intestinal permeability in ApoE−/− mice.
Fig. 7: Intestinal miR-34a/KLF4 signaling regulates atherosclerotic process in ApoE−/− mice.
Fig. 8: Intestinal KLF4/tight junction signaling is closely associated with human atherosclerotic process.

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Acknowledgements

This work was financially supported by Guangdong Basic and Applied Basic Research Foundation (2023A1515030096), Basic Research Fund of Shenzhen Science and Technology Innovation Commission (JCYJ20220530154002006 and JCYJ20220530154003007), National Natural Science Foundation of China (81902113), Medical Scientific Research Foundation of Guangdong Province (A2022308), and Scientific Research Foundation for Young Teacher of Shenzhen University (000002111207).

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  1. These authors contributed equally: He-zhong-rong Nie, Yi-wen Zhou

Authors and Affiliations

  1. Center of clinical laboratory, Shenzhen Hospital, Southern Medical University, Shenzhen, 518000, China

    He-zhong-rong Nie, Yi-wen Zhou & Xiao-hong Yu

  2. Department of Neurology, Affiliated Hangzhou First People’s Hospital, Zhejiang University School of Medicine, Hangzhou, 310003, China

    Cong-guo Yin & Ling-fei Li

  3. Department of Pathophysiology, School of Basic Medical Sciences, Shenzhen University Medical School, Shenzhen, 518000, China

    Hui-qin Hao, Tao Yuan & Yong Pan

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Contributions

HZRN, XHY, HQH and TY performed the experiments and analyzed data. HZRN, YWZ, CGY, and LFL discussed data. HZRN drafted the manuscript. YWZ guided the clinical experiments, and YP designed, guided the whole study and edited manuscript.

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Correspondence to Yong Pan.

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Nie, Hzr., Zhou, Yw., Yu, Xh. et al. Intestinal epithelial Krüppel-like factor 4 alleviates endotoxemia and atherosclerosis through improving NF-κB/miR-34a-mediated intestinal permeability. Acta Pharmacol Sin (2024). https://doi.org/10.1038/s41401-024-01238-3

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