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Danhong Injection Up-regulates miR-125b in Endothelial Exosomes and Attenuates Apoptosis in Post-Infarction Myocardium

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Abstract

Objective

To investigate the involvement of endothelial cells (ECs)-derived exosomes in the anti-apoptotic effect of Danhong Injection (DHI) and the mechanism of DHI-induced exosomal protection against postinfarction myocardial apoptosis.

Methods

A mouse permanent myocardial infarction (MI) model was established, followed by a 14-day daily treatment with DHI, DHI plus GW4869 (an exosomal inhibitor), or saline. Phosphate-buffered saline (PBS)-induced ECs-derived exosomes were isolated, analyzed by miRNA microarray and validated by droplet digital polymerase chain reaction (ddPCR). The exosomes induced by DHI (DHI-exo), PBS (PBS-exo), or DHI+GW4869 (GW-exo) were isolated and injected into the peri-infarct zone following MI. The protective effects of DHI and DHI-exo on MI hearts were measured by echocardiography, Masson’s trichrome staining, and TUNEL apoptosis assay. The Western blotting and quantitative reverse transcription PCR (qRT-PCR) were used to evaluate the expression levels of miR-125b/p53-mediated pathway components, including miR-125b, p53, Bak, Bax, and caspase-3 activities.

Results

DHI significantly improved cardiac function and reduced infarct size in MI mice (P<0.01), which was abolished by the GW4869 intervention. DHI promoted the exosomal secretion in ECs (P<0.01). According to the results of exosomal miRNA microarray assay, 30 differentially expressed miRNAs in the DHI-exo were identified (28 up-regulated miRNAs and 2 down-regulated miRNAs). Among them, DHI significantly elevated miR-125b level in DHI-exo and DHI-treated ECs, a recognized apoptotic inhibitor impeding p53 signaling (P<0.05). Remarkably, treatment with DHI and DHI-exo attenuated apoptosis, elevated miR-125b expression level, inhibited capsase-3 activity, and down-regulated the expression levels of proapoptotic effectors (p53, Bak, and Bax) in post-MI hearts, whereas these effects were blocked by GW4869 (P<0.05 or P<0.01).

Conclusion

DHI and DHI-induced exosomes inhibited apoptosis, promoted the miR-125b expression level, and regulated the p53 apoptotic pathway in post-infarction myocardium.

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Author information

Authors and Affiliations

  1. Department of Cardiology, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, 100010, China

    Si-nai Li, Zi-hao Liu, Ming-xue Zhou, Wei-hong Liu, Xiao-lei Lai, Ping Li, Lei Zhang, Ju-ju Shang, Sheng-lei Qiu, Yan Lou, Yu-pei Tan, Wen-long Xing &  Hong-xu Liu

  2. Beijing Institute of Chinese Medicine, Beijing, 100010, China

    Si-nai Li, Ming-xue Zhou, Wei-hong Liu, Ping Li & Lei Zhang

Authors
  1. Si-nai Li
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  2. Zi-hao Liu
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Contributions

Li SN and Xing WL equally contributed to this work as co-first author. Liu HX made substantial contributions to the conception or design of the work. Li SN and Xing WL made contributions to the acquisition, analysis or interpretation of data for this work and drafted the work. Liu ZH, Zhou MX, LIU WH, Lai XL, Li P, and Zhang L made contributions to the data analysis in Western blot and RT-PCR. Shang JJ, Qiu SL, Lou Y, and Tan YP revised the work for important intellectual content. All authors have agreed with the content and approve of the manuscript for submission.

Corresponding author

Correspondence to Hong-xu Liu.

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The authors declare that there are no conflicts of interest regarding the publication of this paper.

Additional information

Supported by National Natural Science Foundation of China (No. 82174106 and No. 81703850)

Supplementary Materials

11655_2023_3647_MOESM1_ESM.pdf

Appendix 1. Total ion chromatograms of DHI in positive (A) and negative ion mode (B) based on UPLC-TOF MS. P1: Protocatechualdehyde; P2: Salvianolic acid D; P3: Kaempferide; P4: Salvianolic acid A; P5: Coumalic acid; P6: Rosmarinic acid; P7: Salvianolic acid B.

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Li, Sn., Liu, Zh., Zhou, Mx. et al. Danhong Injection Up-regulates miR-125b in Endothelial Exosomes and Attenuates Apoptosis in Post-Infarction Myocardium. Chin. J. Integr. Med. 29, 1099–1110 (2023). https://doi.org/10.1007/s11655-023-3647-7

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