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

[1]	DEVARAJ E, PERUMAL E, SUBRAMANIYAN R, et al. Liver fibrosis: Extracellular vesicles mediated intercellular communication in perisinusoidal space[J]. Hepatology, 2022, 76(1): 275-285. DOI: 10.1002/hep.32239.
[2]	XUE R, FAN JG. Brief introduction of an international expert consensus statement: A new definition of metabolic associated fatty liver disease[J]. J Clin Hepatol, 2020, 36(6): 1224-1227. DOI: 10.3969/j.issn.1001-5256.2020.06.007. 薛芮, 范建高. 代谢相关脂肪性肝病新定义的国际专家共识简介[J]. 临床肝胆病杂志, 2020, 36(6): 1224-1227. DOI: 10.3969/j.issn.1001-5256.2020.06.007.
[3]	ESCUDERO-CASAO M, CARDONA A, BELTRAN-DEBON R, et al. Fluorinated triazole-containing sphingosine analogues. Syntheses and in vitro evaluation as SPHK inhibitors[J]. Org Biomol Chem, 2018, 16(39): 7230-7235. DOI: 10.1039/c8ob01867g.
[4]	ANDERSON AK, LAMBERT JM, MONTEFUSCO DJ, et al. Depletion of adipocyte sphingosine kinase 1 leads to cell hypertrophy, impaired lipolysis, and nonalcoholic fatty liver disease[J]. J Lipid Res, 2020, 61(10): 1328-1340. DOI: 10.1194/jlr.RA120000875.
[5]	AVNI D, HARIKUMAR KB, SANYAL AJ, et al. Deletion or inhibition of SphK1 mitigates fulminant hepatic failure by suppressing TNFα-dependent inflammation and apoptosis[J]. FASEB J, 2021, 35(3): e21415. DOI: 10.1096/fj.202002540R.
[6]	LI L, WANG H, ZHANG J, et al. SPHK1 deficiency protects mice from acetaminophen-induced ER stress and mitochondrial permeability transition[J]. Cell Death Differ, 2020, 27(6): 1924-1937. DOI: 10.1038/s41418-019-0471-x.
[7]	GENG T, SUTTER A, HARLAND MD, et al. SphK1 mediates hepatic inflammation in a mouse model of NASH induced by high saturated fat feeding and initiates proinflammatory signaling in hepatocytes[J]. J Lipid Res, 2015, 56(12): 2359-2371. DOI: 10.1194/jlr.M063511.
[8]	LI Q, QIAN J, LI Y, et al. Generation of sphingosine-1-phosphate by sphingosine kinase 1 protects nonalcoholic fatty liver from ischemia/reperfusion injury through alleviating reactive oxygen species production in hepatocytes[J]. Free Radic Biol Med, 2020, 159: 136-149. DOI: 10.1016/j.freeradbiomed.2020.07.004.
[9]	CHEN Y, AZAD MB, GIBSON SB. Methods for detecting autophagy and determining autophagy-induced cell death[J]. Can J Physiol Pharmacol, 2010, 88(3): 285-295. DOI: 10.1139/Y10-010.
[10]	LAVIEU G, SCARLATTI F, SALA G, et al. Regulation of autophagy by sphingosine kinase 1 and its role in cell survival during nutrient starvation[J]. J Biol Chem, 2006, 281(13): 8518-8527. DOI: 10.1074/jbc.M506182200.
[11]	HUANG LS, NATARAJAN V. Sphingolipids in pulmonary fibrosis[J]. Adv Biol Regul, 2015, 57: 55-63. DOI: 10.1016/j.jbior.2014.09.008.
[12]	CHERESH P, KIM SJ, HUANG LS, et al. The sphingosine kinase 1 inhibitor, PF543, mitigates pulmonary fibrosis by reducing lung epithelial cell mtDNA damage and recruitment of fibrogenic monocytes[J]. Int J Mol Sci, 2020, 21(16): 5595. DOI: 10.3390/ijms21165595.
[13]	HA AW, SUDHADEVI T, EBENEZER DL, et al. Neonatal therapy with PF543, a sphingosine kinase 1 inhibitor, ameliorates hyperoxia-induced airway remodeling in a murine model of bronchopulmonary dysplasia[J]. Am J Physiol Lung Cell Mol Physiol, 2020, 319(3): L497-L512. DOI: 10.1152/ajplung.00169.2020.
[14]	IMBERT C, MONTFORT A, FRAISSE M, et al. Resistance of melanoma to immune checkpoint inhibitors is overcome by targeting the sphingosine kinase-1[J]. Nat Commun, 2020, 11(1): 437. DOI: 10.1038/s41467-019-14218-7.
[15]	SUKOCHEVA OA, FURUYA H, NG ML, et al. Sphingosine kinase and sphingosine-1-phosphate receptor signaling pathway in inflammatory gastrointestinal disease and cancers: A novel therapeutic target[J]. Pharmacol Ther, 2020, 207: 107464. DOI: 10.1016/j.pharmthera.2019.107464.
[16]	JIN LM, HUANG DS, LIU YX, et al. The effects of sphingosine kinase/1-phosphate sphingosine signaling on hepatic ischemia-reperfusion injury in rats[J]. Chin Exp Surg, 2020, 37(10): 1826-1829. DOI: 10.3760/cma.j.cn421213-20191031-00772. 金丽明, 黄东胜, 刘原兴, 等. 鞘氨醇激酶及1-磷酸鞘氨醇信号参与大鼠肝窦微循环保护[J]. 中华实验外科杂志, 2020, 37(10): 1826-1829. DOI: 10.3760/cma.j.cn421213-20191031-00772.
[17]	JIN LM, LIU YX, CHENG J, et al. The effect of SphK1/S1P signaling pathway on hepatic sinus microcirculation in rats with hepatic ischemia-reperfusion injury[J]. Hepatobiliary Pancreat Dis Int, 2022, 21(1): 94-98. DOI: 10.1016/j.hbpd.2021.06.003.
[18]	HUANG LS, SUDHADEVI T, FU P, et al. Sphingosine kinase 1/S1P signaling contributes to pulmonary fibrosis by activating Hippo/YAP pathway and mitochondrial reactive oxygen species in lung fibroblasts[J]. Int J Mol Sci, 2020, 21(6): 2064. DOI: 10.3390/ijms21062064.
[19]	KONO Y, NISHIUMA T, NISHIMURA Y, et al. Sphingosine kinase 1 regulates differentiation of human and mouse lung fibroblasts mediated by TGF-beta1[J]. Am J Respir Cell Mol Biol, 2007, 37(4): 395-404. DOI: 10.1165/rcmb.2007-0065OC.
[20]	CENCETTI F, BERNACCHIONI C, NINCHERI P, et al. Transforming growth factor-beta1 induces transdifferentiation of myoblasts into myofibroblasts via up-regulation of sphingosine kinase-1/S1P3 axis[J]. Mol Biol Cell, 2010, 21(6): 1111-1124. DOI: 10.1091/mbc.e09-09-0812.
[21]	LIAO YS, BAI LL. Role of exercise-induced autophagy in prevention and treatment of nonalcoholic fatty liver disease[J]. J Clin Hepatol, 2022, 38(5): 1156-1160. DOI: 10.3969/j.issn.1001-5256.2022.05.038. 廖粤生, 白莉莉. 运动诱导细胞自噬在非酒精性脂肪性肝病防治中的作用[J]. 临床肝胆病杂志, 2022, 38(5): 1156-1160. DOI: 10.3969/j.issn.1001-5256.2022.05.038.
[22]	LIU GJ, PENG L. Study on the role of autophagy in hepatic fibrosis[J]. Shandong Med J, 2018, 58(48): 108-110. DOI: 10.3969/j.issn.1002-266X.2018.48.031. 刘国菊, 彭雷. 自噬在肝纤维化中作用的研究进展[J]. 山东医药, 2018, 58(48): 108-110. DOI: 10.3969/j.issn.1002-266X.2018.48.031.
[23]	WANG X, WU R, LIU Y, et al. m6A mRNA methylation controls autophagy and adipogenesis by targeting Atg5 and Atg7[J]. Autophagy, 2020, 16(7): 1221-1235. DOI: 10.1080/15548627.2019.1659617.
[24]	KURAMOTO K, KIM YJ, HONG JH, et al. The autophagy protein Becn1 improves insulin sensitivity by promoting adiponectin secretion via exocyst binding[J]. Cell Rep, 2021, 35(8): 109184. DOI: 10.1016/j.celrep.2021.109184.
[25]	ALEXAKI A, GUPTA SD, MAJUMDER S, et al. Autophagy regulates sphingolipid levels in the liver[J]. J Lipid Res, 2014, 55(12): 2521-2531. DOI: 10.1194/jlr.M051862.

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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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