Unacylated ghrelin attenuates acute liver injury and hyperlipidemia via its anti-inflammatory and anti-oxidative activities

Document Type : Original Article

Authors

1 Department of Pharmacy, College of Chemical Engineering, Qingdao University of Science and Technology, Qingdao, China

2 Department of Pathology, Feicheng Hospital Affiliated to Shandong First Medical University, Qingdao, China

3 Department of Pharmacy, Qingdao Women and Children’s Hospital, Qingdao, China

4 Department of Pharmacy College of Chemical Engineering Qingdao University of Science and Technology Qingdao, 266042 China

Abstract

Objective(s): Liver injury and hyperlipidemia are major issues that have drawn more and more attention in recent years. The present study aimed to investigate the effects of unacylated ghrelin (UAG) on acute liver injury and hyperlipidemia in mice.
Materials and Methods: UAG was injected intraperitoneally once a day for three days. Three hours after the last administration, acute liver injury was induced by intraperitoneal injection of carbon tetrachloride (CCl4), and acute hyperlipidemia was induced by intraperitoneal injection of poloxamer 407, respectively. Twenty-four hours later, samples were collected for serum biochemistry analysis, histopathological examination, and Western blotting.
Results: In acute liver injury mice, UAG significantly decreased liver index, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α), reduced malondialdehyde (MDA) concentration and increased superoxide dismutase(SOD) in liver tissue. NF-kappa B (NF-κB) protein expression in the liver was down-regulated. In acute hyperlipidemia mice, UAG significantly decreased serum total cholesterol (TC), triglyceride (TG), ALT, and AST, as well as hepatic TG levels. Meanwhile, hepatic MDA decreased and SOD increased significantly. Moreover, UAG improved the pathological damage in the liver induced by CCl4 and poloxamer 407, respectively.
Conclusion: Intraperitoneal injection of UAG exhibited hepatoprotective and lipid-lowering effects on acute liver injury and hyperlipidemia, which is attributed to its anti-inflammatory and anti-oxidant activities. 

Keywords

Main Subjects


1. Domitrović R, Jakovac H, Milin C, Radosević-Stasić B. Dose- and time-dependent effects of luteolin on carbon tetrachloride-induced hepatotoxicity in mice. Exp Toxicol Pathol 2009; 61:581-589.
2. Mistry S, Dutt KR, Jena J. Protective effect of Sida cordata leaf extract against CCl4 induced acute liver toxicity in rats. Asian Pac J Trop Med 2013; 6:280-284.
3. Mundi MS, Velapati S, Patel J, Kellogg TA, Abu Dayyeh BK, Hurt RT. Evolution of NAFLD and its management. Nutr Clin Pract 2020; 35:72-84. 
4. Ravan AP, Bahmani M, Ghasemi Basir HR, Salehi I, Oshaghi EA. Hepatoprotective effects of Vaccinium arctostaphylos against CCl4-induced acute liver injury in rats. J Basic Clin Physiol Pharmacol 2017; 28:463-471.
5. Stewart J, McCallin T, Martinez J, Chacko S, Yusuf S. Hyperlipidemia. Pediatr Rev 2020; 41:393-402. 
6. Peng Q, Yao X, Xiang J, Wang Y, Lin X. Acupuncture for hyperlipidemia: protocol for a systematic review and meta-analysis. Medicine (Baltimore) 2018; 97:1-4.
7. Poornima IG, Indaram M, Ross JD, Agarwala A, Wild RA. Hyperlipidemia and risk for preclampsia. J Clin Lipidol 2022; 16:253-260. 
8. Wu Z, Han M, Chen T, Yan W, Ning Q. Acute liver failure: mechanisms of immune-mediated liver injury. Liv Int 2010; 30:782-794.
9. Liu B, Fang Y, Yi R, Zhao X. Preventive effect of blueberry extract on liver injury induced by carbon tetrachloride in mice. Foods 2019; 8:48-61.
10. Liu X, Guo Y, Li Z, Gong Y. The role of acylated ghrelin and unacylated ghrelin in the blood and hypothalamus and their interaction with nonalcoholic fatty liver disease. Iran J Basic Med Sci 2020; 23:1191-1196.
11. Ezquerro S, Mocha F, Frühbeck G, Guzmán-Ruiz R, Valentí V, Mugueta C, et al. Ghrelin reduces TNF-α-induced human hepatocyte apoptosis, autophagy, and pyroptosis: role in obesity-associated NAFLD. J Clin Endocrinol Metab 2019; 104:21-37. 
12. Chen CY, Asakawa A, Fujimiya M, Lee SD, Inui A. Ghrelin gene products and the regulation of food intake and gut motility. Pharmacol Rev 2009; 61:430-481.
13. Quiñones M, Fernø J, Al-Massadi O. Ghrelin and liver disease. Rev Endocr Metab Disord 2020; 21:45-56.
14. Raghay K, Akki R, Bensaid D, Errami M. Ghrelin as an anti-inflammatory and protective agent in ischemia/reperfusion injury. Peptides 2020; 124:170226-170267. 
15. Delhanty PJ, Van der Lely AJ. Ghrelin and glucose homeostasis. Peptides 2011; 32:2309-2318.
16. Lewiński A, Karbownik-Lewińska M, Wieczorek-Szukała K, Stasiak M, Stawerska R. Contribution of ghrelin to the pathogenesis of growth hormone deficiency. Int J Mol Sci 2021; 22:9066-9087. 
17. Hilgendorf KI, Johnson CT, Mezger A, Rice SL, Norris AM, Demeter J, et al. Omega-3 fatty acids activate ciliary FFAR4 to control adipogenesis. Cell 2019; 179:1289-1305. 
18. Jin T, Xu Q, Liu X, Huang J, Guo Y, Li Y, et al. Effect of calcium-sensitive receptor agonist R568 on gastric motility and the underlying mechanism. Neuroendocrinology 2022; 113:289-303.
19. Shimada T, Furuta H, Doi A, Ariyasu H, Kawashima H, Wakasaki H, et al. Des-acyl ghrelin protects microvascular endothelial cells from oxidative stress-induced apoptosis through sirtuin 1 signaling pathway. Metabolism 2014; 63:469-474.
20. Ugwu FN, Yu AP, Sin TK, Tam BT, Lai CW, Wong SC, et al. Protective effect of unacylated ghrelin on compression-induced skeletal muscle injury mediated by SIRT1-signaling. Front Physiol 2017; 8:962-975.
21. Alharbi S. Exogenous administration of unacylated ghrelin attenuates hepatic steatosis in high-fat diet-fed rats by modulating glucose homeostasis, lipogenesis, oxidative stress, and endoplasmic reticulum stress. Biomed Pharmacother 2022; 151:113095. 
22. Gortan Cappellari G, Barazzoni R. Ghrelin forms in the modulation of energy balance and metabolism. Eat Weight Disord 2019; 24:997-1013.
23. Ronchi G, Tos P, Angelino E, Muratori L, Reano S, Filigheddu N, et al. Effect of unacylated ghrelin on peripheral nerve regeneration. Eur J Histochem 2021; 65:3287-3294.
24. Au CC, Docanto MM, Zahid H, Raffaelli FM, Ferrero RL, Furness JB, et al. Des-acyl ghrelin inhibits the capacity of macrophages to stimulate the expression of aromatase in breast adipose stromal cells. J Steroid Biochem Mol Biol 2017; 170:49-53.
25. Ali M, Hussain H, Hussain A, Rauf A, Hussain W, Ullah M, et al. Hepatoprotective screening of seriphidium kurramense (Qazilb.) Y.R. Ling. BioMed Res Int 2021; 9026731:1-11.
26. Sila A, Kamoun Z, Ghlissi Z, Makni M, Nasri M, Sahnoun Z, et al. Ability of natural astaxanthin from shrimp by-products to attenuate liver oxidative stress in diabetic rats. Pharmacol Rep 2015; 67:310-316.
27. Xu L, Yu Y, Sang R, Li J, Ge B, Zhang X. Protective effects of taraxasterol against ethanol-induced liver injury by regulating CYP2E1/Nrf2/HO-1 and NF-κB signaling pathways in mice. Oxid Med Cell Longev 2018; 8284107:1-11.
28. Jing ZT, Liu W, Xue CR, Wu SX, Chen WN, Lin XJ, et al. AKT activator SC79 protects hepatocytes from TNF-α-mediated apoptosis and alleviates d-Gal/LPS-induced liver injury. Am J Physiol Gastrointest liver physiol 2019; 316:387-396.
29. Guo Y, Gao S, Jiang Z, Huang J, He X, Jin R, et al. Sun, calcium-sensing receptor (CaSR) agonist R568 inhibits small intestinal motility of mice through neural and non-neural mechanisms. Food Funct 2021; 12:11926-11937.
30. Shukla V, Kaushal JB, Sankhwar P, Manohar M, Dwivedi A. Inhibition of TPPP3 attenuates β-catenin/NF-κB/COX-2 signaling in endometrial stromal cells and impairs decidualization. J Endocrinol 2019; 240:417-429.
31. Somensi N, Rabelo TK, Guimarães AG, Quintans-Junior LJ, Souza Araújo AA, Moreira JCF, et al. Carvacrol suppresses LPS-induced pro-inflammatory activation in RAW 264.7 macrophages through ERK1/2 and NF-kB pathway. Int Immunopharmacol 2019; 75:105743-105750.
32. Yenmis G, Yaprak Sarac E, Besli N, Soydas T, Tastan C, Dilek Kancagi D, et al. Anti-cancer effect of metformin on the metastasis and invasion of primary breast cancer cells through mediating NF-kB activity. Acta Histochem 2021; 123:151709-151719.
33. Lu Y, Hu D, Ma S, Zhao X, Wang S, Wei G, et al. Protective effect of wedelolactone against CCl4-induced acute liver injury in mice. Int Immunopharmacol 2016; 34:44-52.
34. Verhelst K, Carpentier I, Beyaert R. Regulation of TNF-induced NF-κB activation by different cytoplasmic ubiquitination events. Cytokine Growth Factor Rev 2011; 22:277-286.
35. Bragg DA, Walling A. Metabolic syndrome: hyperlipidemia. FP Essent 2015; 435:17-23.
36. Zhang T, Zhao Q, Xiao X, Yang R, Hu D, Zhu X, et al. Modulation of lipid metabolism by celastrol. J Proteome Res 2019; 18:1133-1144. 
37. Younossi ZM, Koenig AB, Abdelatif D, Fazel Y, Henry L, Wymer M. Global epidemiology of nonalcoholic fatty liver disease-meta-analytic assessment of prevalence, incidence, and outcomes. Hepatology 2016; 64:73-84.
38. Omari-Siaw E, Wang Q, Sun C, Gu Z, Zhu Y, Cao X, et al. Tissue distribution and enhanced in vivo anti-hyperlipidemic-anti-oxidant effects of perillaldehyde-loaded liposomal nanoformulation against poloxamer 407-induced hyperlipidemia. Int J Pharm 2016; 513:68-77.
39. Bartelt A, Weigelt C, Cherradi ML, Niemeier A, Tödter K, Heeren J, et al. Effects of adipocyte lipoprotein lipase on de novo lipogenesis and white adipose tissue browning. Biochim Biophys Acta 2013; 1831:934-942.
40. Duivenvoorden I, Teusink B, Rensen PC, Romijn JA, Havekes LM, Voshol PJ. Apolipoprotein C3 deficiency results in diet-induced obesity and aggravated insulin resistance in mice. Diabetes 2005; 54:664-671.
41. Nolan CJ, Prentki M. Insulin resistance and insulin hypersecretion in the metabolic syndrome and type 2 diabetes: time for a conceptual framework shift. Diab Vasc Dis Res 2019; 16:118-127.