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Autophagy of hepatic stellate cell induced by Clonorchis sinensis

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Abstract

Background

Clonorchis sinensis was a food-borne zoonotic parasite in the worldwide and also an important risk factor of hepatic fibrosis. Excretory/secretion products of C. sinensis (CsESPs) are involved in parasite-host interactions and contribute to the development of hepatic damage. The aim of the present study was to investigate whether CsESPs and CsTP (adult protein) could induce autophagy of hepatic stellate cells (HSCs) and further activate HSCs so as to participate in the pathogenesis of hepatic fibrosis.

Methods and results

The human hepatic stellate cell line LX-2 was stimulated by CsESPs and CsTP. CsESPs showed the effect on cell proliferation in methyl thiazolyl tetrazolium (MTT) assay while CsTP failed. Autophagosomes and autolysosomes were observed after the transmission mRFP-EGFP-LC3 plasmid into the LX-2 cells. CsESPs had more powerful to induce the accumulation of autophagosomes and autolysosomes to enhance autophagic flux compared with CsTP. Western-blotting analysis confirmed that the ratio of LC3-II/I in LX-2 cells was up-regulated after CsESPs treatment for 6 h, which further proved that CsESPs could induce autophagy in LX-2 cells. Meanwhile, q-PCR results showed that the mRNA levels of collagen I, collagen III and α-SMA decreased in LX-2 cells after treatment with autophagy inhibitor chloroquine, whereas they increased when combination with CsESPs.

Conclusions

These results suggested that CsESPs-induced autophagy might be involved in the activation of HSCs, and consequently participate in the pathogenesis of hepatic fibrosis caused by C. sinensis infection.

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Data availability

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

Code availability

Image analysis was performed using ImageJ software. Statistical analyses were performed using SPSS 22.0.

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Acknowledgements

This research was funded by the Natural Science Foundation of Guangxi (No. 2020GXNSFAA159068 and 2019GXNSFAA245069), Guangxi First-class Discipline Project for Basic Medicine (No. GXFCDP-BM-2018), Guangdong Province University Student Innovation and Entrepreneurship Training Programs (No. S201910570080), Guangzhou Medical University Student Science and Technology Innovation Programs (No. 2018A094) China

Funding

This research was funded by the Natural Science Foundation of Guangxi (No. 2020GXNSFAA159068 and 2019GXNSFAA245069), Guangxi First-class Discipline Project for Basic Medicine (No. GXFCDP-BM-2018), Guangdong Province University Student Innovation and Entrepreneurship Training Programs (No. S201910570080), Guangzhou Medical University Student Science and Technology Innovation Programs (No. 2018A094).

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Author notes

  1. Bao Zheng and Zhiyan Gao equally contributed to this study.

Authors and Affiliations

  1. Department of Pathogen Biology & Immunology, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, 511436, People’s Republic of China

    Jingying Mai & Changling Ma

  2. Department of Parasitology, School of Basic Medical Sciences, Guangxi Medical University, Nanning, 530021, People’s Republic of China

    Bao Zheng, Yunyu Lu & Yanwen Li

  3. Department of Morphology Experiment Center, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, 511436, People’s Republic of China

    Zhiyan Gao

  4. KingMed College of Laboratory Medicine, Guangzhou Medical University, Guangzhou, 511436, People’s Republic of China

    Liumei Liang, Yongting Kong, Wanting Chen, Keying Lin & Wanqi Chen

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  1. Bao Zheng
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Contributions

All authors contributed to the study. Conception, design, manuscript modification, and team coordination, etc. (CM and YL), Methodology (BZ, ZG, LL, YL, YK, and WC); Material preparation, data collection and analysis (KL, WC, JM); The first draft of the manuscript (BZ and ZG). All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yanwen Li or Changling Ma.

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All animal procedures in this study were approved by the Animal Care & Welfare Committee of Guangxi Medical University.

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Zheng, B., Gao, Z., Liang, L. et al. Autophagy of hepatic stellate cell induced by Clonorchis sinensis. Mol Biol Rep 49, 1895–1902 (2022). https://doi.org/10.1007/s11033-021-07001-9

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