Exosome-derived miR-1275 mediates IL-38 upregulation in lymphocytes to suppress lipopolysaccharide-induced apoptosis of myocardial cells in vitro

doi:10.12122/j.issn.1673-4254.2025.08.05

Abstract

Abstract:

Objective To investigate the effect of cardiomyocytes-derived exosomes on lipopolysaccharide (LPS)-induced cardiomyocyte injury and its mechanism. Methods Exosomes isolated from rat cardiomyocytes with or without LPS treatment were co-cultured with rat lymphocytes. The lymphocytes with or without exosome treatment were co-cultured with LPS-induced rat cardiomyocytes for 48 h. Cardiomyocyte apoptosis was detected using flow cytometry, and the expressions of apoptosis marker proteins and the PI3K/AKT pathway proteins were detected using Western blotting. The effects of human recombinant IL-38 protein on apoptosis and protein expressions in LPS-induced cardiomyocytes were examined. Results Compared with normal cardiomyocyte-derived exosomes, the exosomes from LPS-induced cardiomyocytes significantly enhanced proliferation and increased mRNA and protein expression levels of IL-38 in rat lymphocytes. Bioinformatics analysis suggested that miR-1275 in the exosome played a key role in LPS-induced cardiomyocyte injury, and in dual luciferase reporter gene assay, miR-1275 mimics significantly increased luciferase activity of WT-IL-38. Co-culture with lymphocytes treated with exosomes from LPS-induced cardiomyocytes significantly inhibited apoptosis of LPS-induced cardiomyocytes. Treatment with recombinant IL-38 also effectively lowered apoptosis rate of LPS-induced cardiomyocytes, reduced cellular expression of Bax protein, and increased the protein expression levels of Bcl-2, p-PI3K and p-AKT. Conclusion miR-1275 in exosomes derived from LPS-induced cardiomyocytes mediates IL-38 up-regulation expression in lymphocytes to activate the PI3K/AKT pathway and inhibit LPS-induced cardiomyocyte apoptosis.

Key words: exosomes, cardiomyocyte, miR-1275, IL-38, sepsis

Haimei BO, Xinying CAO, Pingchuan XING, Zhijun WANG. Exosome-derived miR-1275 mediates IL-38 upregulation in lymphocytes to suppress lipopolysaccharide-induced apoptosis of myocardial cells in vitro[J]. Journal of Southern Medical University, 2025, 45(8): 1608-1615.

Figures/Tables 6

Fig.1 Expression of miRNAs in exosomes detected by qRT-PCR. *P<0.05 vs miR-556 group.

Fig.2 LPS-induced cardiomyocyte injury model. A: Effect of LPS on proliferation of cardiomyocytes. B: Volcano map. C: Heat map. *P<0.05 vs control group.

Fig.3 Effect of exosomes containing miR-1275 on lymphocyte proliferation and mRNA and protein expression levels of IL-38. A: Western blotting for detecting expressions of exosome markers CD63 and TSG101. B: CCK-8 assay for assessing the effect of exosomes on lymphocyte proliferation. C: Expression level of IL-38 protein in lymphocytes detected by Western blotting. Group ① was the control group and group ② was the LPS-exosome group. D: Expression level of IL-38 mRNA in lymphocytes detected by qRT-PCR. E: Dual-luciferase reporter gene assay of the regulatory relationship between miR-1275 and IL-38. *P<0.05 vs control group.

Fig.4 Flow cytometry for assessing of the effect of LPS-induced cardiomyocyte-derived exosomes on cardiomyocyte apoptosis. *P<0.05 vs control group.

Fig.5 Effect of IL-38 on cardiomyocyte apoptosis detected by flow cytometry. *P<0.05 vs control group.

Fig.6 Western blotting for assessing the effect of IL-38-mediated PI3K/AKT pathway on exosomes-induced cardiomyocyte apoptosis. A: Expression levels of apoptosis-related proteins Bax and Bcl-2. B: Expression levels of proteins related to the PI3K/AKT pathway. ① Control group; ② Re-IL-38 group. *P<0.05 vs control group.

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