Mechanism of action of sphingosine kinase 1 inhibitor in a rat model of liver fibrosis

Yan WANG; Li MA; Xiong XIAO; Dongmei LEI; Fengru ZHAO; Liyuan GUO; Xiaozhong WANG

doi:10.3969/j.issn.1001-5256.2023.08.018

Volume 39 Issue 8

Aug. 2023

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Article Navigation > Journal of Clinical Hepatology > 2023 > 39(8): 1886-1894

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Mechanism of action of sphingosine kinase 1 inhibitor in a rat model of liver fibrosis

DOI: 10.3969/j.issn.1001-5256.2023.08.018

1.
The Fifth People's Hospital Affiliated to Chengdu University of Traditional Chinese Medicine, Chengdu 611130, China
2.
Xinjiang Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Urumqi 830000, China
3.
Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region, Urumqi 830000, China

Research funding:

National Natural Science Foundtion of China (81860808)

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Corresponding author: WANG Yan, wy751105@126.com (ORCID: 0000-0001-9730-8279)
Received Date: 2022-11-25
Accepted Date: 2023-01-19
Published Date: 2023-08-20

Abstract

Abstract

Objective To investigate the therapeutic effect of sphingosine kinase 1 (SphK1) inhibitor in different liver fibrosis models. Methods A total of 170 Sprague-Dawley rats were randomly divided into normal group (30 rats), HFE group (45 rats given high-fat emulsion), CCl₄ group (45 rats induced by CCl₄), and CCl₄+HFE group (50 rats given HFE and CCl₄). After successful modeling confirmed by pathology and laboratory examination, the SphK1 inhibitor PF-543 was given, and the rats in the same group were given normal saline as control. Liver tissue samples were collected on days 1, 7, and 14 for Masson staining; the percentage of liver fibrosis area was compared; the formation of autophagosome was observed under a transmission electron microscope; real-time PCR was used to measure the mRNA expression levels of markers associated with liver fibrosis and autophagy. A one-way analysis of variance was used for comparison between multiple groups, and the LSD method was used for further comparison between two groups; a Spearman correlation analysis was also performed. Results Compared with the normal group, the CCl₄ group and the HFE+CCl₄ group had a significant increase in the percentage of liver fibrosis area (6.93±5.81/10.89±2.67 vs 0.57±0.13, both P < 0.01), the CCl₄ group and the HFE+CCl₄ group had a significant reduction in the percentage of liver fibrosis area on day 7 (both P < 0.01). Compared with the normal group, the HFE group, the CCl₄ group, and the HFE+CCl₄ group had a significant reduction in the expression level of SphK1 (all P < 0.01), and the HFE+CCl₄ group had significant increases in the mRNA expression levels of alpha-smooth muscle actin, transforming growth factor-β, and collagen type Ⅰ alpha 1 (all P < 0.01), while there were no significant changes in these indices after intervention (all P > 0.05). The expression levels of Atg5, Atg12, and Becn1 were negatively correlated with the area of liver fibrosis (r=-0.715, -0.640, and -0.632, all P < 0.01), and the expression level of SphK1 was positively correlated with the mRNA expression of Atg5, Atg12, Becn1, and Map1lc3a (r=0.603, 0.561, 0.510, and 0.498, all P < 0.01). Electron microscopy showed that the CCL₄ group had slight edema, abundant organelles, and mild swelling in liver tissue, mainly the expansion of rough endoplasmic reticulum, and nine autolysosome (ASS) structures were seen in this field, while no typical ASS structure was observed on day 7 of PF-543 intervention. The HFE+CCL₄ group had mild edema, unclear structure, and abundant rough endoplasmic reticulum with marked expansion and few ribosome particles attached to its surface in liver tissue, and 6 ASS structures were seen in this field, while no typical ASS structure was observed on day 7 of PF-543 intervention. Conclusion PF-543 significantly inhibits autophagy and is associated with a reduction in fibrosis area. It is suggested that targeting SphK1 can affect the level of liver autophagy, thereby alleviating the state of liver fibrosis in two liver fibrosis models.
- Hepatic Fibrosis,
- Sphingosine Kinase 1,
- Autophagy,
- Models, Animal

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References

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Mechanism of action of sphingosine kinase 1 inhibitor in a rat model of liver fibrosis

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Mechanism of action of sphingosine kinase 1 inhibitor in a rat model of liver fibrosis

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