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Autophagic degradation of MVBs in LSECs promotes Aldosterone induced-HSCs activation

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Abstract

Background and aims

The important role of extracellular vesicles (EVs) in liver fibrosis has been confirmed. However, EVs derived from liver sinusoidal endothelial cells (LSECs) in the activation of hepatic stellate cells (HSCs) and liver fibrosis is still unclear. Our previous work demonstrated that Aldosterone (Aldo) may have the potential to regulate EVs from LSECs via autophagy pathway. Thus, we aim to investigate the role of Aldo in the regulation of EVs derived from LSECs.

Approach and results

Using an Aldo-continuous pumping rat model, we observed that Aldo-induced liver fibrosis and capillarization of LSECs. In vitro, transmission electron microscopy (TEM) revealed that stimulation of Aldo led to the upregulation of autophagy and degradation of multivesicular bodies (MVBs) in LSECs. Mechanistically, Aldo upregulated ATP6V0A2, which promoted lysosomal acidification and subsequent autophagy in LSECs. Inhibiting autophagy with si-ATG5 adeno-associated virus (AAV) in LSECs effectively mitigated Aldo-induced liver fibrosis in rats. RNA sequencing and nanoparticle tracking (NTA) analyses of EVs derived from LSECs indicated that Aldo result in a decrease in both the quantity and quality of EVs. We also observed a reduction in the protective miRNA-342-5P in EVs derived from Aldo-treated LSECs, which may play a critical role in HSCs activation. Target knockdown of EV secretion with si-RAB27a AAV in LSECs led to the development of liver fibrosis and HSC activation in rats.

Conclusion

Aldo-induced Autophagic degradation of MVBs in LSECs promotes a decrease in the quantity and quality of EVs derived from LSECs, resulting in the activation of HSCs and liver fibrosis under hyperaldosteronism. Modulating the autophagy level of LSECs and their EV secretion may represent a promising therapeutic approach for treating liver fibrosis.

Graphical abstract

In a physiological state, LSECs transmit inhibitory signals to HSCs via the secretion of EVs that are rich in miR-342-5p. However, in pathological conditions, the elevated levels of serum aldosterone induce capillarization and excessive autophagy in LSECs. This autophagy leads to the degradation of MVBs in LSECs, resulting in a reduction of the number of EVs and miR-342-5p content within EVs. This reduction ultimately leads to a diminished inhibitory signal transmitted to HSCs, thereby activating HSCs and promoting the development of liver fibrosis.

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Data Availability

The RNA sequencing Data of EVs derived from LSECs with or without Aldo stimulation of this study are openly available in NCBI SRA, reference BioProject ID PRJNA982445.

Abbreviations

LSEC:

Liver sinusoidal endothelial cell

HSC:

Hepatic stellate cell

Aldo:

Aldosterone

Spi:

Spironolactone

MR:

Mineralocorticoid receptor

RAAS:

Renin–Angiotensin–Aldosterone system

NO:

Nitric oxide

MVB:

Multivesicular body

EV:

Extracellular vesicle

AAV:

Adeno-associated virus

STAB2:

Stabling 2

ATG5:

Autophagy-related 5

ATP6V0A2:

ATPase H + transporting V0 subunit a2

CD31:

Platelet endothelial cell adhesion molecule 1

LC3B:

Microtubule-associated protein 1 light chain 3 beta

TEM:

Transmission electron microscopy

vWF:

Von Willebrand Factor

Col I:

Collagen I

αSMA:

α-Smooth muscle actin

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Funding

This work was supported by the National Natural Science Foundation of China (grant number 81670556 and 82170641 to Xu Li), Guangdong Science and Technology Project (grant number 2017B020209003 and 2021A1515012595 to Xu Li), The Youth Fund of National Natural Science Foundation of China (grant number 82100660 to Yang Li, 81700541 to Zuowei Ning), and China postdoctoral Science Foundation (grant number 2021M691458 to Yang Li).

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Author notes

  1. Tingting Chen, Yan Zhang, Yijie Zhang and Zuowei Ning are contributed equally to this work.

Authors and Affiliations

  1. Department of Emergency Medicine, Nanfang Hospital, Southern Medical University, No. 1838, North of Guangzhou Avenue, Guangzhou, 510515, Guangdong, China

    Tingting Chen, Yan Zhang, Yijie Zhang, Qihan Xu, Ying Lin, Jiacheng Gong, Jierui Li, Zhuoer Chen, Yang Li & Xu Li

  2. Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Tingting Chen, Yan Zhang, Yijie Zhang, Zuowei Ning, Qihan Xu, Ying Lin, Jiacheng Gong, Jierui Li & Zhuoer Chen

  3. Department of Respiratory Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Ying Meng

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  1. Tingting Chen
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Contributions

XL, YL, and YM designed and supervised the study. XL, YL and ZN acquired funding. TC and YZ performed the experiments. TC, YZ, YZ, and YL analyzed the data. JW, JL, JG, and QX collected samples and clinical information. TC drafted the manuscript. XL, YL, and YM reviewed the manuscript.

Corresponding authors

Correspondence to Ying Meng, Yang Li or Xu Li.

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Conflict of interest

Tingting Chen, Yan Zhang, Yijie Zhang, Zuowei Ning, Qihan Xu, Ying Lin, Jiacheng Gong, Jierui Li, Zhuoer Chen, Ying Meng, Yang Li and Xu Li declares have no conflict of interest.

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Chen, T., Zhang, Y., Zhang, Y. et al. Autophagic degradation of MVBs in LSECs promotes Aldosterone induced-HSCs activation. Hepatol Int 18, 273–288 (2024). https://doi.org/10.1007/s12072-023-10559-0

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