Lactoferrin Enhanced Apoptosis and Protected Against Thioacetamide-Induced Liver Fibrosis in Rats

Alyaa Hessin; Rehab Hegazy; Azza Hassan; Nemat Yassin; Sanaa Kenawy

doi:10.3889/oamjms.2015.038

Authors

Alyaa Hessin National Research Center, Medical Division, Pharmacology Department, Giza
Rehab Hegazy National Research Center, Medical Division, Pharmacology Department, Giza
Azza Hassan Cairo University, Faculty of Veterinary Medicine, Pathology Department, Giza
Nemat Yassin National Research Center, Medical Division, Pharmacology Department, Giza
Sanaa Kenawy Cairo University, Faculty of Pharmacy, Pharmacology and Toxicology Department, Cairo

DOI:

https://doi.org/10.3889/oamjms.2015.038

Keywords:

Liver fibrosis, Lactoferrin, Thioacetamide, Apoptosis, NF-Îº B, Alpha fetoprotein

Abstract

BACKGROUND: Liver fibrosis is the common pathologic consequence of all chronic liver diseases.

AIM: Lactoferrin (Lf) was investigated for its possible hepatoprotective effect against thioacetamide (TAA)-induced liver fibrosis rat model.

MATERIAL AND METHODS: Rats received TAA (200 mg/kg/biweekly, ip) for four successive weeks. Lf (200 mg/kg/day, p.o.) or vehicle (VHC) was administered for one month before and another month during TAA injection. Body weight and mortality rate were assessed during the month of TAA-intoxication. Thereafter, serum and liver tissues were analyzed for liver function, oxidative, fibrotic and apoptotic markers.

RESULTS: Lf conserved rats against TAA-induced body weight-loss and mortality. Preservation of serum albumin, alkaline phosphatase and total bilirubin levels was also observed. Lf also protected rats against TAA-induced decrease in reduced glutathione and increase in malondialdehyde liver contents. Normal liver contents of hydroxyproline, nuclear factor kappa B and alpha fetoprotein; as markers of fibrosis; were increased with TAA and conserved with Lf-TAA. Lf maintained the normal architecture of the liver and immunohistochemical findings revealed increase in apoptotic bodies compared to TAA that favored necrosis.

CONCLUSION: In conclusion, Lf improved liver function, reduced oxidative stress and liver fibrosis, and enhanced apoptosis in rats with liver fibrosis, suggesting it to have useful therapeutic potential in patients with liver fibrosis.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

Plum Analytics Artifact Widget Block

References

Liu T, Wang X, Karsdal MA, Leeming DJ, Genovese F. Molecular serum markers of liver fibrosis. Biomarker insights. 2012;7:105-17. DOI: https://doi.org/10.4137/BMI.S10009

Alshawsh MA, Abdulla MA, Ismail S, Amin ZA. Hepatoprotective Effects of Orthosiphon stamineus Extract on Thioacetamide-Induced Liver Cirrhosis in Rats. Evidence-based complementary and alternative medicine : eCAM. 2011;2011:103039. DOI: https://doi.org/10.1155/2011/103039

Mengoli M, Parmeggiani D, Mengoli MC, Grinzi G, Tolomelli S. [Drug-induced hepatotoxicity: clinical and biochemical features of 26 patients and a review of the literature]. Recenti progressi in medicina. 2011;102(6):253-60.

Papamichalis PA, Zachou K, Koukoulis GK, Veloni A, Karacosta EG, Kypri L, et al. The revised international autoimmune hepatitis score in chronic liver diseases including autoimmune hepatitis/overlap syndromes and autoimmune hepatitis with concurrent other liver disorders. Journal of autoimmune diseases. 2007;4:3. DOI: https://doi.org/10.1186/1740-2557-4-3

Przybylkowski A, Gromadzka G, Chabik G, Wierzchowska A, Litwin T, Czlonkowska A. Liver cirrhosis in patients newly diagnosed with neurological phenotype of Wilson's disease. Functional neurology. 2014:1-7. DOI: https://doi.org/10.11138/FNeur/2014.29.1.023

Moreira RK. Hepatic stellate cells and liver fibrosis. Archives of pathology & laboratory medicine. 2007;131(11):1728-34. DOI: https://doi.org/10.5858/2007-131-1728-HSCALF

Ramadori G, Moriconi F, Malik I, Dudas J. Physiology and pathophysiology of liver inflammation, damage and repair. Journal of physiology and pharmacology : an official journal of the Polish Physiological Society. 2008;59 Suppl 1:107-17.

Henderson NC, Iredale JP. Liver fibrosis: cellular mechanisms of progression and resolution. Clin Sci (Lond). 2007;112(5):265-80. DOI: https://doi.org/10.1042/CS20060242

Ahmad A, Ahmad R. Understanding the mechanism of hepatic fibrosis and potential therapeutic approaches. Saudi journal of gastroenterology : official journal of the Saudi Gastroenterology Association. 2012;18(3):155-67. DOI: https://doi.org/10.4103/1319-3767.96445

Harn HJ, Lin SZ, Hung SH, Subeq YM, Li YS, Syu WS, et al. Adipose-derived stem cells can abrogate chemical-induced liver fibrosis and facilitate recovery of liver function. Cell transplantation. 2012;21(12):2753-64. DOI: https://doi.org/10.3727/096368912X652959

Friedman SL. Evolving challenges in hepatic fibrosis. Nature reviews Gastroenterology & hepatology. 2010;7(8):425-36. DOI: https://doi.org/10.1038/nrgastro.2010.97

Zhao W, Li JJ, Cao DY, Li X, Zhang LY, He Y, et al. Intravenous injection of mesenchymal stem cells is effective in treating liver fibrosis. World journal of gastroenterology : WJG. 2012;18(10):1048-58. DOI: https://doi.org/10.3748/wjg.v18.i10.1048

Levay PF, Viljoen M. Lactoferrin: a general review. Haematologica. 1995;80(3):252-67.

Berlutti F, Pantanella F, Natalizi T, Frioni A, Paesano R, Polimeni A, et al. Antiviral properties of lactoferrin--a natural immunity molecule. Molecules. 2011;16(8):6992-7018. DOI: https://doi.org/10.3390/molecules16086992

Gonzalez-Chavez SA, Arevalo-Gallegos S, Rascon-Cruz Q. Lactoferrin: structure, function and applications. International journal of antimicrobial agents. 2009;33(4):301 e1-8. DOI: https://doi.org/10.1016/j.ijantimicag.2008.07.020

Velusamy SK, Poojary R, Ardeshna R, Alabdulmohsen W, Fine DH, Velliyagounder K. Protective effects of human lactoferrin during Aggregatibacter actinomycetemcomitans-induced bacteremia in lactoferrin-deficient mice. Antimicrobial agents and chemotherapy. 2014;58(1):397-404. DOI: https://doi.org/10.1128/AAC.00020-13

Carvalho CA, Sousa IP, Jr., Silva JL, Oliveira AC, Goncalves RB, Gomes AM. Inhibition of Mayaro virus infection by bovine lactoferrin. Virology. 2014;452-453:297-302. DOI: https://doi.org/10.1016/j.virol.2014.01.022

Fang B, Zhang M, Tian M, Jiang L, Guo HY, Ren FZ. Bovine lactoferrin binds oleic acid to form an anti-tumor complex similar to HAMLET. Biochimica et biophysica acta. 2014;1841(4):535-43. DOI: https://doi.org/10.1016/j.bbalip.2013.12.008

Shi H, Li W. Inhibitory effects of human lactoferrin on U14 cervical carcinoma through upregulation of the immune response. Oncology letters. 2014;7(3):820-6. DOI: https://doi.org/10.3892/ol.2013.1776

Gifford JL, Hunter HN, Vogel HJ. Lactoferricin: a lactoferrin-derived peptide with antimicrobial, antiviral, antitumor and immunological properties. Cellular and molecular life sciences : CMLS. 2005;62(22):2588-98. DOI: https://doi.org/10.1007/s00018-005-5373-z

Yin H, Cheng L, Holt M, Hail N, Jr., Maclaren R, Ju C. Lactoferrin protects against acetaminophen-induced liver injury in mice. Hepatology. 2010;51(3):1007-16. DOI: https://doi.org/10.1002/hep.23476

Vesce F, Giugliano E, Bignardi S, Cagnazzo E, Colamussi C, Marci R, et al. Vaginal Lactoferrin Administration before Genetic Amniocentesis Decreases Amniotic Interleukin-6 Levels. Gynecologic and obstetric investigation. 2014. DOI: https://doi.org/10.1159/000358877

Tung YT, Chen HL, Yen CC, Lee PY, Tsai HC, Lin MF, et al. Bovine lactoferrin inhibits lung cancer growth through suppression of both inflammation and expression of vascular endothelial growth factor. Journal of dairy science. 2013;96(4):2095-106. DOI: https://doi.org/10.3168/jds.2012-6153

Abramovitch S, Dahan-Bachar L, Sharvit E, Weisman Y, Ben Tov A, Brazowski E, et al. Vitamin D inhibits proliferation and profibrotic marker expression in hepatic stellate cells and decreases thioacetamide-induced liver fibrosis in rats. Gut. 2011;60(12):1728-37. DOI: https://doi.org/10.1136/gut.2010.234666

Bruck R, Genina O, Aeed H, Alexiev R, Nagler A, Avni Y, et al. Halofuginone to prevent and treat thioacetamide-induced liver fibrosis in rats. Hepatology. 2001;33(2):379-86. DOI: https://doi.org/10.1053/jhep.2001.21408

Togawa J, Nagase H, Tanaka K, Inamori M, Umezawa T, Nakajima A, et al. Lactoferrin reduces colitis in rats via modulation of the immune system and correction of cytokine imbalance. American journal of physiology Gastrointestinal and liver physiology. 2002;283(1):G187-95. DOI: https://doi.org/10.1152/ajpgi.00331.2001

Cocchetto DM, Bjornsson TD. Methods for vascular access and collection of body fluids from the laboratory rat. Journal of pharmaceutical sciences. 1983;72(5):465-92. DOI: https://doi.org/10.1002/jps.2600720503

Woessner JF, Jr. The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Archives of biochemistry and biophysics. 1961;93:440-7. DOI: https://doi.org/10.1016/0003-9861(61)90291-0

Carleton HM, Drury RAB, Wallington EA. Carleton's Histological technique. 5th ed ed. Oxford: Oxford University Press; 1980.

Klainguti M, Aigner S, Kilo J, Eppenberger HM, Mandinova A, Aebi U, et al. Lack of nuclear apoptosis in cardiomyocytes and increased endothelin-1 levels in a rat heart model of myocardial stunning. Basic research in cardiology. 2000;95(4):308-15. DOI: https://doi.org/10.1007/s003950070050

Kadir FA, Kassim NM, Abdulla MA, Yehye WA. Hepatoprotective Role of Ethanolic Extract of Vitex negundo in Thioacetamide-Induced Liver Fibrosis in Male Rats. Evidence-based complementary and alternative medicine : eCAM. 2013;2013:739850. DOI: https://doi.org/10.1155/2013/739850

Li X, Benjamin IS, Alexander B. Reproducible production of thioacetamide-induced macronodular cirrhosis in the rat with no mortality. Journal of hepatology. 2002;36(4):488-93. DOI: https://doi.org/10.1016/S0168-8278(02)00011-9

Kadir FA, Othman F, Abdulla MA, Hussan F, Hassandarvish P. Effect of Tinospora crispa on thioacetamide-induced liver cirrhosis in rats. Indian journal of pharmacology. 2011;43(1):64-8. DOI: https://doi.org/10.4103/0253-7613.75673

Sakka SG. Assessing liver function. Current opinion in critical care. 2007;13(2):207-14. DOI: https://doi.org/10.1097/MCC.0b013e328012b268

Al-Attar AM. Hepatoprotective influence of vitamin C on thioacetamide-induced liver cirrhosis in Wistar male rats. J Pharmacol Toxicol. 2011; 6(3):218-33. DOI: https://doi.org/10.3923/jpt.2011.218.233

Salama SM, Abdulla MA, Alrashdi AS, Hadi AH. Mechanism of Hepatoprotective Effect of Boesenbergia rotunda in Thioacetamide-Induced Liver Damage in Rats. Evidence-based complementary and alternative medicine : eCAM. 2013;2013:157456. DOI: https://doi.org/10.1155/2013/157456

Gomez HF, Ochoa TJ, Herrera-Insua I, Carlin LG, Cleary TG. Lactoferrin protects rabbits from Shigella flexneri-induced inflammatory enteritis. Infection and immunity. 2002;70(12):7050-3. DOI: https://doi.org/10.1128/IAI.70.12.7050-7053.2002

Hoekstra LT, de Graaf W, Nibourg GA, Heger M, Bennink RJ, Stieger B, et al. Physiological and biochemical basis of clinical liver function tests: a review. Annals of surgery. 2013;257(1):27-36. DOI: https://doi.org/10.1097/SLA.0b013e31825d5d47

Galisteo M, Suarez A, del Pilar Montilla M, del Pilar Utrilla M, Jimenez J, Gil A, et al. Antihepatotoxic activity of Rosmarinus tomentosus in a model of acute hepatic damage induced by thioacetamide. Phytotherapy research : PTR. 2000;14(7):522-6. DOI: https://doi.org/10.1002/1099-1573(200011)14:7<522::AID-PTR660>3.0.CO;2-K

Ueno H, Sato T, Yamamoto S, Tanaka K, Ohkawa S, Takagi H, et al. Randomized, double-blind, placebo-controlled trial of bovine lactoferrin in patients with chronic hepatitis C. Cancer science. 2006;97(10):1105-10. DOI: https://doi.org/10.1111/j.1349-7006.2006.00274.x

Yamaguchi M, Matsuura M, Kobayashi K, Sasaki H, Yajima T, Kuwata T. Lactoferrin protects against development of hepatitis caused by sensitization of Kupffer cells by lipopolysaccharide. Clinical and diagnostic laboratory immunology. 2001;8(6):1234-9. DOI: https://doi.org/10.1128/CDLI.8.6.1234-1239.2001

Guerra RR, Trotta MR, Aloia TPA, Dagli MLZ, Hernandez-Blazquez FJ. A novel chronic cirrhosis TAA-induced model in rats. Brazilian J Vet Pathol. 2010;3(1):9-16.

Al-Jumaily EF, Khaleel FM. The effect of chronic liver diseases on some biochemical parameters in patients serum. Curr Res J Biol Sci. 2012;4(5):638-42.

Shirin H, Sharvit E, Aeed H, Gavish D, Bruck R. Atorvastatin and rosuvastatin do not prevent thioacetamide induced liver cirrhosis in rats. World journal of gastroenterology : WJG. 2013;19(2):241-8. DOI: https://doi.org/10.3748/wjg.v19.i2.241

Stankova P, Kucera O, Lotkova H, Rousar T, Endlicher R, Cervinkova Z. The toxic effect of thioacetamide on rat liver in vitro. Toxicology in vitro : an international journal published in association with BIBRA. 2010;24(8):2097-103. DOI: https://doi.org/10.1016/j.tiv.2010.06.011

Pallottini V, Martini C, Bassi AM, Romano P, Nanni G, Trentalance A. Rat HMGCoA reductase activation in thioacetamide-induced liver injury is related to an increased reactive oxygen species content. Journal of hepatology. 2006;44(2):368-74. DOI: https://doi.org/10.1016/j.jhep.2005.06.011

Okuyama H, Nakamura H, Shimahara Y, Araya S, Kawada N, Yamaoka Y, et al. Overexpression of thioredoxin prevents acute hepatitis caused by thioacetamide or lipopolysaccharide in mice. Hepatology. 2003;37(5):1015-25. DOI: https://doi.org/10.1053/jhep.2003.50203

Hong SW, Jung KH, Zheng HM, Lee HS, Suh JK, Park IS, et al. The protective effect of resveratrol on dimethylnitrosamine-induced liver fibrosis in rats. Archives of pharmacal research. 2010;33(4):601-9. DOI: https://doi.org/10.1007/s12272-010-0415-y

Latorre D, Puddu P, Valenti P, Gessani S. Reciprocal interactions between lactoferrin and bacterial endotoxins and their role in the regulation of the immune response. Toxins. 2010;2(1):54-68. DOI: https://doi.org/10.3390/toxins2010054

Ogasawara Y, Imase M, Oda H, Wakabayashi H, Ishii K. Lactoferrin directly scavenges hydroxyl radicals and undergoes oxidative self-degradation: a possible role in protection against oxidative DNA damage. International journal of molecular sciences. 2014;15(1):1003-13. DOI: https://doi.org/10.3390/ijms15011003

Lv P, Luo HS, Zhou XP, Xiao YJ, Paul SC, Si XM, et al. Reversal effect of thalidomide on established hepatic cirrhosis in rats via inhibition of nuclear factor-kappaB/inhibitor of nuclear factor-kappaB pathway. Archives of medical research. 2007;38(1):15-27. DOI: https://doi.org/10.1016/j.arcmed.2006.09.006

Ribeiro PS, Cortez-Pinto H, Sola S, Castro RE, Ramalho RM, Baptista A, et al. Hepatocyte apoptosis, expression of death receptors, and activation of NF-kappaB in the liver of nonalcoholic and alcoholic steatohepatitis patients. The American journal of gastroenterology. 2004;99(9):1708-17. DOI: https://doi.org/10.1111/j.1572-0241.2004.40009.x

Elsharkawy AM, Mann DA. Nuclear factor-kappaB and the hepatic inflammation-fibrosis-cancer axis. Hepatology. 2007;46(2):590-7. DOI: https://doi.org/10.1002/hep.21802

Park JH, Kum YS, Lee TI, Kim SJ, Lee WR, Kim BI, et al. Melittin attenuates liver injury in thioacetamide-treated mice through modulating inflammation and fibrogenesis. Exp Biol Med (Maywood). 2011;236(11):1306-13. DOI: https://doi.org/10.1258/ebm.2011.011127

Breitkopf K, Sawitza I, Westhoff JH, Wickert L, Dooley S, Gressner AM. Thrombospondin 1 acts as a strong promoter of transforming growth factor beta effects via two distinct mechanisms in hepatic stellate cells. Gut. 2005;54(5):673-81. DOI: https://doi.org/10.1136/gut.2004.042911

Chen Y, Yang Y, Miller ML, Shen D, Shertzer HG, Stringer KF, et al. Hepatocyte-specific Gclc deletion leads to rapid onset of steatosis with mitochondrial injury and liver failure. Hepatology. 2007;45(5):1118-28. DOI: https://doi.org/10.1002/hep.21635

Bastway Ahmed M, Hasona N, Selemain A. Protective effects of extract from dates (phoenix dactylifera l.) And ascorbic acid on thioacetamide-induced hepatotoxicity in rats. Iranian journal of pharmaceutical research : IJPR. 2008;7(3):193-201.

Hori Y, Sato S, Yamate J, Kurasaki M, Nishihira J, Hosokawa T, et al. Immunohistochemical study of macrophage migration inhibitory factor in rat liver fibrosis induced by thioacetamide. European journal of histochemistry : EJH. 2003;47(4):317-24. DOI: https://doi.org/10.4081/842

Chavez E, Reyes-Gordillo K, Segovia J, Shibayama M, Tsutsumi V, Vergara P, et al. Resveratrol prevents fibrosis, NF-kappaB activation and TGF-beta increases induced by chronic CCl4 treatment in rats. Journal of applied toxicology : JAT. 2008;28(1):35-43. DOI: https://doi.org/10.1002/jat.1249

Ikeda M, Nozaki A, Sugiyama K, Tanaka T, Naganuma A, Tanaka K, et al. Characterization of antiviral activity of lactoferrin against hepatitis C virus infection in human cultured cells. Virus research. 2000;66(1):51-63. DOI: https://doi.org/10.1016/S0168-1702(99)00121-5

Iwasa M, Kaito M, Ikoma J, Takeo M, Imoto I, Adachi Y, et al. Lactoferrin inhibits hepatitis C virus viremia in chronic hepatitis C patients with high viral loads and HCV genotype 1b. The American journal of gastroenterology. 2002;97(3):766-7. DOI: https://doi.org/10.1111/j.1572-0241.2002.05573.x

Wei M, Xu Y, Zou Q, Tu L, Tang C, Xu T, et al. Hepatocellular carcinoma targeting effect of PEGylated liposomes modified with lactoferrin. European journal of pharmaceutical sciences: official journal of the European Federation for Pharmaceutical Sciences. 2012; 46(3):131-41. DOI: https://doi.org/10.1016/j.ejps.2012.02.007