Abstract
The development of acute liver injury can result in liver cirrhosis, liver failure, and even liver cancer, yet there is currently no effective therapy for it. The purpose of this study was to investigate the protective effect and therapeutic mechanism of Lycium barbarum polysaccharides (LBPs) on acute liver injury induced by carbon tetrachloride (CCl4). To create a model of acute liver injury, experimental canines received an intraperitoneal injection of 1 mL/kg of CCl4 solution. The experimental canines in the therapy group were then fed LBPs (20 mg/kg). CCl4-induced liver structural damage, excessive fibrosis, and reduced mitochondrial density were all improved by LBPs, according to microstructure data. By suppressing Kelch-like epichlorohydrin (ECH)-associated protein 1 (Keap1), promoting the production of sequestosome 1 (SQSTM1)/p62, nuclear factor erythroid 2-related factor 2 (Nrf2), and phase II detoxification genes and proteins downstream of Nrf2, and restoring the activity of anti-oxidant enzymes like catalase (CAT), LBPs can restore and increase the antioxidant capacity of liver. To lessen mitochondrial damage, LBPs can also enhance mitochondrial respiration, raise tissue adenosine triphosphate (ATP) levels, and reactivate the respiratory chain complexes I–V. According to serum metabolomics, the therapeutic impact of LBPs on acute liver damage is accomplished mostly by controlling the pathways to lipid metabolism. 9-Hydroxyoctadecadienoic acid (9-HODE), lysophosphatidylcholine (LysoPC/LPC), and phosphatidylethanolamine (PE) may be potential indicators of acute liver injury. This study confirmed that LBPs, an effective hepatoprotective drug, may cure acute liver injury by lowering oxidative stress, repairing mitochondrial damage, and regulating metabolic pathways.
概要
急性肝损伤的发展可导致肝硬化、 肝衰竭, 甚至肝癌, 但目前尚无有效的治疗方法. 本研究旨在探讨枸杞多糖 (LBPs) 对四氯化碳 (CCl4) 诱导的急性肝损伤的保护作用及其治疗机制. 为了建立急性肝损伤模型, 以1 mL/kg的剂量对实验犬进行CCl4溶液的腹腔注射. 然后给治疗组的实验犬喂食枸杞多糖 (20 mg/kg). 微观结果显示, CCl4诱导的肝脏结构损伤、 过度纤维化和线粒体密度降低都可以通过给予LBPs得到改善. LBPs可以通过抑制Kelch样ECH相关蛋白1 (Keap1), 促进蛋白 (p62)、 核因子E2相关因子2 (Nrf2)和Nrf2下游的II期解毒基因和蛋白的产生, 以及恢复过氧化氢酶 (CAT) 等抗氧化酶的活性, 恢复并增加肝脏的抗氧化能力. LBPs还可以增强线粒体呼吸能力, 提高组织ATP水平, 以及重新激活呼吸链复合体I–V以减轻线粒体的损伤. 血清代谢组学结果显示, LBPs对急性肝损伤的治疗作用主要通过控制脂质代谢途径实现. 9-羟基十八碳二烯酸 (9-HODE)、 溶血磷脂酰胆碱 (LPC) 和磷脂酰乙醇胺 (PE) 可能是急性肝损伤的潜在标志物. 这项研究证实, LBPs是一种有效的保肝药物, 可以通过降低氧化应激、 修复线粒体损伤和调节代谢途径来治疗急性肝损伤.
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References
Balogun KA, Albert CJ, Ford DA, et al., 2013. Dietary omega-3 polyunsaturated fatty acids alter the fatty acid composition of hepatic and plasma bioactive lipids in C57BL/6 mice: a lipidomic approach. PLoS ONE, 8(11):e82399. https://doi.org/10.1371/journal.pone.0082399
Bellezza I, Giambanco I, Minelli A, et al., 2018. Nrf2-Keap1 signaling in oxidative and reductive stress. Biochim Biophys Acta Mol Cell Res, 1865(5):721–733. https://doi.org/10.1016/j.bbamcr.2018.02.010
Brand MD, Nicholls DG, 2011. Assessing mitochondrial dysfunction in cells. Biochem J, 435(2):297–312. https://doi.org/10.1042/BJ20110162
Caldez MJ, van Hul N, Koh HWL, et al., 2018. Metabolic remodeling during liver regeneration. Dev Cell, 47(4):425–438.e5. https://doi.org/10.1016/j.devcel.2018.09.020
Calzada E, Avery E, Sam PN, et al., 2019. Phosphatidylethanolamine made in the inner mitochondrial membrane is essential for yeast cytochrome bc1 complex function. Nat Commun, 10:1432. https://doi.org/10.1038/s41467-019-09425-1
Chen CF, Wang K, Zhang HF, et al., 2019. A unique SUMO-interacting motif of Trx2 is critical for its mitochondrial presequence processing and anti-oxidant activity. Front Physiol, 10:1089. https://doi.org/10.3389/fphys.2019.01089
Chen YP, Liu KH, Zhang JW, et al., 2020. c-Jun NH2-terminal protein kinase phosphorylates the Nrf2-ECH homology 6 domain of nuclear factor erythroid 2-related factor 2 and downregulates cytoprotective genes in acetaminophen-induced liver injury in mice. Hepatology, 71(5):1787–1801. https://doi.org/10.1002/hep.31116
Du X, Zhang JJ, Liu L, et al., 2022. A novel anticancer property of Lycium barbarum polysaccharide in triggering ferroptosis of breast cancer cells. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(4):286–299. https://doi.org/10.1631/jzus.B2100748
Elvas F, Stroobants S, Wyffels L, 2017. Phosphatidylethanol-amine targeting for cell death imaging in early treatment response evaluation and disease diagnosis. Apoptosis, 22(8):971–987. https://doi.org/10.1007/s10495-017-1384-0
Frazier AE, Vincent AE, Turnbull DM, et al., 2020. Assessment of mitochondrial respiratory chain enzymes in cells and tissues. Methods Cell Biol, 155:121–156. https://doi.org/10.1016/bs.mcb.2019.11.007
Gao J, Feng ZH, Wang XQ, et al., 2018. SIRT3/SOD2 maintains osteoblast differentiation and bone formation by regulating mitochondrial stress. Cell Death Differ, 25(2):229–240. https://doi.org/10.1038/cdd.2017.144
Gonzalez E, van Liempd S, Conde-Vancells J, et al., 2012. Serum UPLC-MS/MS metabolic profiling in an experimental model for acute-liver injury reveals potential biomarkers for hepatotoxicity. Metabolomics, 8(6):997–1011. https://doi.org/10.1007/s11306-011-0329-9
Gou ZX, Su XJ, Hu X, et al., 2020. Melatonin improves hypoxic-ischemic brain damage through the Akt/Nrf2/Gpx4 signaling pathway. Brain Res Bull, 163:40–48. https://doi.org/10.1016/j.brainresbull.2020.07.011
Guo Q, Zhang QQ, Chen JQ, et al., 2017. Liver metabolomics study reveals protective function of Phyllanthus urinaria against CCl4-induced liver injury. Chin J Nat Med, 15(7): 525–533. https://doi.org/10.1016/S1875-5364(17)30078-X
Haga S, Yimin, Ozaki M, 2017. Relevance of FXR-p62/SQSTM1 pathway for survival and protection of mouse hepatocytes and liver, especially with steatosis. BMC Gastroenterol, 17:9. https://doi.org/10.1186/s12876-016-0568-3
Hinkovska-Galcheva V, Treadwell T, Shillingford JM, et al., 2021. Inhibition of lysosomal phospholipase A2 predicts drug-induced phospholipidosis. J Lipid Res, 62:100089. https://doi.org/10.1016/j.jlr.2021.100089
Hu W, Dang XB, Wang G, et al., 2018. Peroxiredoxin-3 attenuates traumatic neuronal injury through preservation of mitochondrial function. Neurochem Int, 114:120–126. https://doi.org/10.1016/j.neuint.2018.02.004
Jain A, Lamark T, Sjøttem E, et al., 2010. p62/SQSTM1 is a target gene for transcription factor NRF2 and creates a positive feedback loop by inducing antioxidant response element-driven gene transcription. J Biol Chem, 285(29): 22576–22591. https://doi.org/10.1074/jbc.M110.118976
Jia R, Li Y, Cao LP, et al., 2019. Antioxidative, anti-inflammatory and hepatoprotective effects of resveratrol on oxidative stress-induced liver damage in tilapia (Oreochromis niloticus). Comp Biochem Physiol C Toxicol Pharmacol, 215: 56–66. https://doi.org/10.1016/j.cbpc.2018.10.002
Jardim FR, de Almeida FJS, Luckachaki MD, et al., 2020. Effects of sulforaphane on brain mitochondria: mechanistic view and future directions. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(4):263–279. https://doi.org/10.1631/jzus.B1900614
Jin Y, Huang ZL, Li L, et al., 2019. Quercetin attenuates toosendanin-induced hepatotoxicity through inducing the Nrf2/GCL/GSH antioxidant signaling pathway. Acta Pharmacol Sin, 40:75–85. https://doi.org/10.1038/s41401-018-0024-8
Johnson CH, Ivanisevic J, Siuzdak G, 2016. Metabolomics: beyond biomarkers and towards mechanisms. Nat Rev Mol Cell Biol, 17(7):451–459. https://doi.org/10.1038/nrm.2016.25
Kandel J, Angelin AA, Wallace DC, et al., 2016. Mitochondrial respiration is sensitive to cytoarchitectural breakdown. Integr Biol (Camb), 8(11):1170–1182. https://doi.org/10.1039/c6ib00192k
Larsen S, Nielsen J, Hansen CN, et al., 2012. Biomarkers of mitochondrial content in skeletal muscle of healthy young human subjects. J Physiol, 590(14):3349–3360. https://doi.org/10.1113/jphysiol.2012.230185
Lewis KN, Wason E, Edrey YH, et al., 2015. Regulation of Nrf2 signaling and longevity in naturally long-lived rodents. Proc Natl Acad Sci USA, 112(12):3722–3727. https://doi.org/10.1073/pnas.1417566112
Lin SY, Xu D, Du XX, et al., 2019. Protective effects of salidroside against carbon tetrachloride (CCl4)-induced liver injury by initiating mitochondria to resist oxidative stress in mice. Int J Mol Sci, 20(13):3187. https://doi.org/10.3390/ijms20133187
Liu BX, Zeng QW, Chen HM, et al., 2021. The hepatotoxicity of altrazine exposure in mice involves the intestinal microbiota. Chemosphere, 272:129572. https://doi.org/10.1016/j.chemosphere.2021.129572
Liu JZ, Wang X, Liu R, et al., 2014. Oleanolic acid co-administration alleviates ethanol-induced hepatic injury via Nrf-2 and ethanol-metabolizing modulating in rats. Chem Biol Interact, 221:88–98. https://doi.org/10.1016/j.cbi.2014.07.017
Liu WW, Zhou Y, Duan WZ, et al., 2021. Glutathione peroxidase 4-dependent glutathione high-consumption drives acquired platinum chemoresistance in lung cancer-derived brain metastasis. Clin Transl Med, 11(9):e517. https://doi.org/10.1002/ctm2.517
Lustgarten MS, Bhattacharya A, Muller FL, et al., 2012. Complex I generated, mitochondrial matrix-directed superoxide is released from the mitochondria through voltage dependent anion channels. Biochem Biophys Res Commun, 422(3):515–521. https://doi.org/10.1016/j.bbrc.2012.05.055
Ma J, Yu J, Su XR, et al., 2014. UPLC-MS-based serum metabonomics for identifying acute liver injury biomarkers in Chinese miniature pigs. Toxicol Lett, 225(3):358–366. https://doi.org/10.1016/j.toxlet.2014.01.008
Ma Q, 2010. Transcriptional responses to oxidative stress: pathological and toxicological implications. Pharmacol Ther, 125(3):376–393. https://doi.org/10.1016/j.pharmthera.2009.11.004
Maciejewska D, Drozd A, Skonieczna-Żydecka K, et al., 2020. Eicosanoids in nonalcoholic fatty liver disease (NAFLD) progression. Do serum eicosanoids profile correspond with liver eicosanoids content during NAFLD development and progression? Molecules, 25(9):2026. https://doi.org/10.3390/molecules25092026
Myers CR, 2012. The effects of chromium (VI) on the thioredoxin system: implications for redox regulation. Free Radic Biol Med, 52(10):2091–2107. https://doi.org/10.1016/j.freeradbiomed.2012.03.013
Osthues T, Zimmer B, Rimola V, et al., 2020. The lipid receptor G2A (GPR132) mediates macrophage migration in nerve injury-induced neuropathic pain. Cells, 9(7):1740. https://doi.org/10.3390/cells9071740
Qiao N, Yang YY, Liao JZ, et al., 2021. Metabolomics and transcriptomics indicated the molecular targets of copper to the pig kidney. Ecotoxicol Environ Saf, 218:112284. https://doi.org/10.1016/j.ecoenv.2021.112284
Raghunath A, Sundarraj K, Nagarajan R, et al., 2018. Antioxidant response elements: discovery, classes, regulation and potential applications. Redox Biol, 17:297–314. https://doi.org/10.1016/j.redox.2018.05.002
Ralph SJ, Moreno-Sánchez R, Neuzil J, et al., 2011. Inhibitors of succinate: quinone reductase/Complex II regulate production of mitochondrial reactive oxygen species and protect normal cells from ischemic damage but induce specific cancer cell death. Pharm Res, 28(11):2695–2730. https://doi.org/10.1007/s11095-011-0566-7
Rolin J, Al-Jaderi Z, Maghazachi AA, 2013. Oxidized lipids and lysophosphatidylcholine induce the chemotaxis and intracellular calcium influx in natural killer cells. Immunobiology, 218(6):875–883. https://doi.org/10.1016/j.imbio.2012.10.009
Saito K, Maekawa K, Ishikawa M, et al., 2014. Glucosylceramide and lysophosphatidylcholines as potential blood biomarkers for drug-induced hepatic phospholipidosis. Toxicol Sci, 141(2):377–386. https://doi.org/10.1093/toxsci/kfu132
Sharma S, Bhattarai S, Ara H, et al., 2020. SOD2 deficiency in cardiomyocytes defines defective mitochondrial bioenergetics as a cause of lethal dilated cardiomyopathy. Redox Biol, 37:101740. https://doi.org/10.1016/j.redox.2020.101740
Shelton P, Jaiswal AK, 2013. The transcription factor NF-E2-related factor 2 (Nrf2): a protooncogene? FASEB J, 27(2): 414–423. https://doi.org/10.1096/fj.12-217257
Sun HY, Hu YJ, Zhao XY, et al., 2015. Age-related changes in mitochondrial antioxidant enzyme Trx2 and TXNIP-Trx2-ASK1 signal pathways in the auditory cortex of a mimetic aging rat model: changes to Trx2 in the auditory cortex. FEBS J, 282(14):2758–2774. https://doi.org/10.1111/febs.13324
Tian XJ, Liang TS, Liu YL, et al., 2019. Extraction, structural characterization, and biological functions of Lycium barbarum polysaccharides: a review. Biomolecules, 9(9):389. https://doi.org/10.3390/biom9090389
Tu WJ, Wang H, Li S, et al., 2019. The anti-inflammatory and anti-oxidant mechanisms of the Keap1/Nrf2/ARE signaling pathway in chronic diseases. Aging Dis, 10(3):637–651. https://doi.org/10.14336/AD.2018.0513
Vance JE, 2015. Phospholipid synthesis and transport in mammalian cells. Traffic, 16(1):1–18. https://doi.org/10.1111/tra.12230
Venâncio C, Antunes L, Félix L, et al., 2013. Chronic ketamine administration impairs mitochondrial complex I in the rat liver. Life Sci, 93(12–14):464–470. https://doi.org/10.1016/j.lfs.2013.08.001
Wang M, Wang L, Han L, et al., 2017. The effect of carabrone on mitochondrial respiratory chain complexes in Gaeumannomyces graminis. J Appl Microbiol, 123(5): 1100–1110. https://doi.org/10.1111/jam.13554
Wu ZG, Han MF, Chen T, et al., 2010. Acute liver failure: mechanisms of immune-mediated liver injury. Liver Int, 30(6):782–794. https://doi.org/10.1111/j.1478-3231.2010.02262.x
Xin YF, Zhang S, Gu LQ, et al., 2011. Electrocardiographic and biochemical evidence for the cardioprotective effect of antioxidants in acute doxorubicin-induced cardiotoxicity in the beagle dogs. Biol Pharm Bull, 34(10):1523–1526. https://doi.org/10.1248/bpb.34.1523
Xu DW, Xu M, Jeong S, et al., 2019. The role of Nrf2 in liver disease: novel molecular mechanisms and therapeutic approaches. Front Pharmacol, 9:1428. https://doi.org/10.3389/fphar.2018.01428
Zhang JQ, Shi L, Xu XN, et al., 2014. Therapeutic detoxification of quercetin against carbon tetrachloride-induced acute liver injury in mice and its mechanism. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 15(12):1039–1047. https://doi.org/10.1631/jzus.B1400104
Zhang Q, Zhang W, Liu J, et al., 2021. Lysophosphatidylcholine promotes intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression in human umbilical vein endothelial cells via an orphan G protein receptor 2-mediated signaling pathway. Bioengineered, 12(1): 4520–4535. https://doi.org/10.1080/21655979.2021.1956671
Zhao YN, Lu J, Mao AK, et al., 2021. Autophagy inhibition plays a protective role in ferroptosis induced by alcohol via the p62-Keap1-Nrf2 pathway. J Agric Food Chem, 69(33):9671–9683. https://doi.org/10.1021/acs.jafc.1c03751
Acknowledgments
This work was supported by the Science and Technology Project of Shaoguan Science and Technology Bureau (No. 200811104530939). In addition, we would like to thank Na QIAO and Hanming CHEN from the Veterinary Medicine of South China Agricultural University for their help in the experiment and professional advice in the process of writing this manuscript.
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Jianjia HUANG: data curation, investigation, visualization, conceptualization, and writing original draft. Yuman BAI: formal analysis, investigation, and methodology. Wenting XIE: review and data curation. Rongmei WANG: funding acquisition. Wenyue QIU: investigation and conceptualization. Shuilian ZHOU: formal analysis. Zhaoxin TANG: resources. Jianzhao LIAO: writing, reviewing, and editing. Rongsheng SU: project administration, data curation, resources, supervision, writing, reviewing, and editing. All authors have read and approved the final manuscript, and therefore, have full access to all the data in the study and take responsibility for the integrity and security of the data.
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Jianjia HUANG, Yuman BAI, Wenting XIE, Rongmei WANG, Wenyue QIU, Shuilian ZHOU, Zhaoxin TANG, Jianzhao LIAO, and Rongsheng SU declare that they have no conflict of interest.
All institutional and national guidelines for the care and use of laboratory animals were followed. All experimental procedures and animal care were carried out in accordance with pertinent guidelines and regulations approved by the Institutional Animal Care and Use Committee of South China Agricultural University (No. 2020A132).
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Huang, J., Bai, Y., Xie, W. et al. Lycium barbarum polysaccharides ameliorate canine acute liver injury by reducing oxidative stress, protecting mitochondrial function, and regulating metabolic pathways. J. Zhejiang Univ. Sci. B 24, 157–171 (2023). https://doi.org/10.1631/jzus.B2200213
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DOI: https://doi.org/10.1631/jzus.B2200213
Key words
- Acute liver injury
- Oxidative stress
- Mitochondrial dysfunction
- Metabolomics
- Lycium barbarum polysaccharides