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Catalpol inhibits hepatic stellate cell activation by reducing the formation and changing the contents of hepatocyte-derived extracellular vesicles

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Journal of Cell Communication and Signaling Aims and scope

Abstract

Hepatic stellate cell (HSC) activation is the central event in hepatic fibrosis. The cross-talk between HSCs and hepatocytes, which is mediated by extracellular vesicles (EVs), affects HSC activation. This study aimed to investigate whether Catalpol (CTP) attenuated hepatic fibrosis via modulating EVs. Mice were injected intraperitoneally with CCl4 for 4 weeks to induce hepatic fibrosis. They were gavaged with CTP daily. Mouse serum EVs were isolated and identified using nanoparticle tracking analysis and transmission electron microscopy. Mouse hepatocytes (AML12) and primary HSCs were used to investigate the cell-to-cell crosstalk. The autophagosome-autolysosome fusion was determined using the autophagic flux assay. Hepatic fibrosis was attenuated by CTP, with a decrease of the myofibroblast marker, alpha-smooth muscle actin. The CTP treatment lowered the serum EVs. The co-culture of HSCs and the EVs derived from the CTP-treated mice or hepatocytes reduced HSC proliferation and the expressions of ACTA2 and Col1a1. After the CCl4 treatment, the autophagosomes in AML12 cells were increased, while the autolysosomes were reduced. The decrease of autophagic cargo receptor SQSTM1 in the CTP group suggested that autophagic degradation was sustained. After inhibiting the endogenous Rac1-GTP of hepatocytes, the co-culture of EVs and HSCs reduced Rac1-GTP. The Rac1-GTP level in serum EVs from the CTP-treated mice was reduced in vivo. CTP inhibited autophagy in hepatocytes by reducing Rac1-GTP and thus affect the amount of Rac1-GTP in hepatocyte-derived EVs and the formation of EVs, which attenuated hepatic fibrosis via inhibiting HSC activation.

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Availability of data and material

The datasets used and/or analyzed during the current study are available from the corresponding author upon reasonable request.

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Funding

The study was supported by the National Natural Science Foundation of China [Grant No. 81860119 and 82260131], Key Research and Development Program of Jiangxi Provincial Department of Science and Technology [Grant No. 20203BBG73044], Key Program of Natural Science Foundation of Jiangxi Province of China [Grant No. 20212ACB206017], and Science and Technology Project Foundation of Education Department of Jiangxi Province, China [Grant No. GJJ200193].

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Authors and Affiliations

  1. Department of Gastroenterology, The Second Affiliated Hospital of Nanchang University, No.1 Mingde Road, Nanchang, 330006, Jiangxi Province, People’s Republic of China

    Zheng-Yuan Xie

  2. Medical College of Nanchang University, Nanchang, Jiangxi, China

    Heng-Wei Cao, Qing Wang, Hui Lu & Wen Du

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Correspondence to Zheng-Yuan Xie.

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This study was approved by the Institute Research Medical Ethics Committee of The Second Affiliated Hospital of Nanchang University. All animal experiments complied with the ARRIVE guidelines.

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Supplemental Figure 1

A AML12 cells were transfected with pmCherry-EGFP-LC3b (mouse) plasmid and the autophagic flux assay was performed. The number of autophagosomes (white arrow) and the number of autolysosomes (yellow arrow) were indicated. B CTP affects autophagy in hepatocytes by regulating Rac1 activation and thus affects the amount of activated Rac1 in hepatocyte derived EVs

Supplemental Figure 2

CCl4 (0.5 µl/g) was dissolved in olive oil at 25% (V/V). Balb/c mice were injected intraperitoneally with CCl4 twice a week for 4 weeks. Mice in the CTP group and the Mock group were daily gavaged with 10 mg/kg CTP or 0.9% normal saline after the first injection of CCl4 (n = 5 in each group). Mice were executed at 0 w, 2 w, and 4 w. Mouse liver tissues were fixed and stained using HE

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Xie, ZY., Cao, HW., Wang, Q. et al. Catalpol inhibits hepatic stellate cell activation by reducing the formation and changing the contents of hepatocyte-derived extracellular vesicles. J. Cell Commun. Signal. 17, 723–736 (2023). https://doi.org/10.1007/s12079-022-00716-9

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  • DOI: https://doi.org/10.1007/s12079-022-00716-9

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