MILK WHEY PROTEINS MODULATE ENDOTOXEMIA-INDUCED HEPATOTOXICITY IN RATS
Keywords:
Whey Protein, Alpha-Lactalbumin, Beta-Lactoglobulin, Lipopolysaccharide, Superoxide Dismutase, Total Antioxidant Capacity, Tumor Necrosis Factor-Alpha, Caspase-3, RatsAbstract
Objective: The therapeutic effects of whey protein isolate (WPI), alpha-lactalbumin (a-LA) and beta-lactoglobulin (β-LG) against endotoxemia-induced hepatotoxicity were investigated in female rats.
Methods: Endotoxemia was induced by intraperitoneal injection of Escherichia coli lipopolysaccharides (E. coli LPS) on the 1st day (4 mg/kg) and the 8th day (2 mg/kg) in two weeks-experiment during which two dose levels of WPI, a-LA and β-LG (100 and 200 mg/kg) were administered orally to female rats. At the end of the experiment, sera and liver tissues were collected for biochemical assessment of superoxide dismutase (SOD) activity, DNA-fragmentation, total antioxidant capacity (TAC) and tumor necrosis factor-alpha (TNF-α) level as well as immunohistochemical examination of caspase-3 activity and collagen deposition by Masson's trichome staining.
Results: Endotoxemia, induced by LPS, significantly decreased hepatic SOD enzymatic antioxidant activity as well as serum total antioxidant capacity (TAC). An elevation in hepatic DNA fragmentation and serum levels of tumor necrosis factor-alpha (TNF-α) was also detected. LPS showed strong activation of caspase-3 in perivascular and parenchymal hepatocytes. Abundant bridging of pericentral collagen fibers in hepatotoxic rats was evident in Masson's trichrome staining. Oral treatment with WPI, a-LA and β-LG (100 and 200 mg/kg) inhibited the decrease in hepatic SOD activity and abrogated hepatic DNA damage. Serum levels of TNF-α decreased significantly while serum TAC was improved by WPI, a-LA and β-LG treatment in hepatotoxic groups. These results were confirmed by immune histochemical examination of activated caspase-3 and Masson's trichrome staining of enhanced collagen deposition which were modulated by whey proteins (WPs) after two weeks of treatment.
Conclusion: Milk-derived WPI, a-LA and β-LG can be a tool in combating hepatotoxicity induced by invasive endotoxemia.
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Mehta NN, McGillicuddy FC, Anderson PD, Hinkle CC, Shah R, Pruscino L, et al. Experimental endotoxemia induces adipose inflammation and insulin resistance in humans. Diabetes 2010;59(1):172-81.
Karaa A, Kamoun WS, Clemens MG. Chronic ethanol sensitizes the liver to endotoxin via effects on endothelial nitric oxide synthase regulation. Shock 2005;24(5):447-54.
Dorresteijn MJ, Draisma A, van der Hoeven JG, Pickkers P. Lipopolysaccharide-stimulated whole blood cytokine production does not predict the inflammatory response in human endotoxemia. Innate Immun 2010;16(4):248-53.
Haegens A, Heeringa P, van Suylen RJ, Steele C, Aratani Y, O'Donoghue RJ, et al. Myeloperoxidase deficiency attenuates lipopolysaccharide-induced acute lung inflammation and subsequent cytokine and chemokine production. J Immunol 2009;182(12):7990-6.
Hietbrink F, Besselink MG, Renooij W, de Smet MB, Draisma A, van der Hoeven H, et al. Systemic inflammation increases intestinal permeability during experimental human endotoxemia. Shock 2009;32(4):374-8.
Srivastava A, Maggs JL, Antoine DJ, Williams DP, Smith DA, Park BK. Role of reactive metabolites in drug-induced hepatotoxicity. Handb Exp Pharmacol 2010(196):165-94.
Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. Oxford: Clarendon Press; 1989.
Iszard MB, Liu J, Klaassen CD. Effect of several metallothionein inducers on oxidative stress defense mechanisms in rats. Toxicol 1995;104(1-3):25-33.
Parodi PW. A role for milk proteins in cancer prevention. Aust J Dairy Technol 1998;53:37-47.
Bounous G, Papenburg R, Kongshavn PA, Gold P, Fleiszer D. Dietary whey protein inhibits the development of dimethylhydrazine induced malignancy. Clin Invest Med 1988;11(3):213-7.
Sukkar SG, Bounous UG. The role of whey protein in antioxidant defense. Rivista Italiana di Nutrizione Parenterale ed Enterale 2004;22(4):193-200.
Bounous G. Whey protein concentrate (WPC) and glutathione modulation in cancer treatment. Anticancer Res 2000;20(6C):4785-92.
Yoo YC, Watanabe S, Watanabe R, Hata K, Shimazaki K, Azuma I. Bovine lactoferrin and Lactoferricin inhibit tumor metastasis in mice. Adv Exp Med Biol 1998;443:285-91.
Low PP, Rutherfurd KJ, Gill HS, Cross ML. Effect of dietary whey protein concentrate on primary and secondary antibody responses in immunized BALB/c mice. Int Immunopharmacol 2003;3(3):393-401.
Ajello M, Greco R, Giansanti F, Massucci MT, Antonini G, Valenti P. Anti-invasive activity of bovine lactoferrin towards group A streptococci. Biochem Cell Biol 2002;80(1):119-24.
Weinberg ED. The role of iron in cancer. Eur J Cancer Prev 1996;5(1):19-36.
Kimball SR, Jefferson LS. Control of protein synthesis by amino acid availability. Curr Opin Clin Nutr Metab Care 2002;5(1):63-7.
Mercier A, Gauthier SF, Fliss I. Immunomodulating effects of whey proteins and their enzymatic digests. Int Dairy J 2004;14:175-83.
Gauthier SF, Pouliot Y, Saint-Sauveur D. Imunomodulatory peptides obtained by the enzymatic hydrolysis of whey proteins. Int Dairy J 2006;16:1315-23.
Pearson JM, Bailie MB, Fink GD, Roth RA. Neither platelet activating factor nor leukotrienes are critical mediators of liver injury after lipopolysaccharide administration. Toxicol 1997;121(3):181-9.
Cuzzocrea S, Mazzon E, Di Paola R, Esposito E, Macarthur H, Matuschak GM, et al. A role for nitric oxide-mediated peroxynitrite formation in a model of endotoxin-induced shock. J Pharmacol Exp Ther 2006;319(1):73-81.
Mansour DFS, Eldenshary ES, Nada SA, Omara EA, Ibrahim MIM. Therapeutic effectiveness of certain whey proteins on lipopolysaccharide-induced oxidative stress and histopathological changes in rat liver. J Appl Sci Res 2013;9(8):4983-92.
Kathem RA, El-Denshary ES, Nada SA, El-Yamany M, Mansour DF, Medhat D, et al. Antinociceptive and Anti-Inflammatory activity of bovine Milk-Derived-Lactoglobulin: study of combined effect with tramadol or indomethacin. Global J Pharmacol 2014;8(3):444-56.
Eliwa HA, El-Denshary ES, Nada SA, Elyamany MF, Omara EA, Asaaf N. Evaluation of the therapeutic effect of whey proteins on the hepatotoxicity induced by paracetamol and alcohol coadministration in rats. Int J Pharm Res Bio-Sci 2014;3(2):295-314.
Hamad EM, Taha SH, Abou Dawood AG, Sitohy MZ, Abdel-Hamid M. Protective effect of whey proteins against nonalcoholic fatty liver in rats. Lipids Health Dis 2011;10:57.
Yamaguchi M, Yoshida K, Uchida M. Novel functions of bovine milk-derived alpha-lactalbumin: anti-nociceptive and anti-inflammatory activity caused by inhibiting cyclooxygenase-2 and phospholipase A2. Biol Pharm Bull 2009;32(3):366-71.
Gad AS, Khadrawy YA, El-Nekeety AA, Mohamed SR, Hassan NS, Abdel-Wahhab MA. Antioxidant activity and hepatoprotective effects of whey protein and Spirulina in rats. Nutr 2011;27(5):582-9.
Abdel-Aziem SH, Hassan AM, Abdel-Wahhab MA. Dietary supplementation with whey protein and ginseng extract counteracts oxidative stress and DNA damage in rats fed an aflatoxin-contaminated diet. Mutat Res 2011;723(1):65-71.
De Carvalho-Silva LB, Pacheco MTB, Bertoldo R, Veloso CC, Teodoro LC, Pablo AP, et al. Anti-inflammatory activities of enzymatic (alcalase) hydrolysate of a whey protein concentrate. Afr J Biotechnol 2012;11(12):2993-9.
Cocchetto DM, Bjornsson TD. Methods for vascular access and collection of body fluids from the laboratory rat. J Pharm Sci 1983;72(5):465-92.
Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.
Marklund S, Marklund G. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 1974;47(3):469-74.
Burton K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J 1956;62(2):315-23.
Squier MK, Cohen JJ. Standard quantitative assays for apoptosis. Mol Biotechnol 2001;19(3):305-12.
Koracevic D, Koracevic G, Djordjevic V, Andrejevic S, Cosic V. Method for the measurement of antioxidant activity in human fluids. J Clin Pathol 2001;54(5):356-61.
Morgan RA, Manning PB, Coran AG, Drongowski RA, Till GO, Ward PD, et al. Oxygen free radical activity during live E. coli septic shock in the dog. Circ Shock 1988;25(4):319-23.
Jaeschke H. Reactive oxygen and mechanisms of inflammatory liver injury. J Gastroenterol Hepatol 2000;15(7):718-24.
Spolarics Z. Endotoxemia, pentose cycle, and the oxidant/antioxidant balance in the hepatic sinusoid. J Leukoc Biol 1998;63(5):534-41.
Halliwell B, Gutteridge JM. Role of free radicals and catalytic metal ions in human disease: an overview. Methods Enzymol 1990;186:1-85.
Macdonald J, Galley HF, Webster NR. Oxidative stress and gene expression in sepsis. Br J Anaesth 2003;90(2):221-32.
Zhao L, Chen YH, Wang H, Ji YL, Ning H, Wang SF, et al. Reactive oxygen species contribute to lipopolysaccharide-induced teratogenesis in mice. Toxicol Sci 2008;103(1):149-57.
Hsu DZ, Chiang PJ, Chien SP, Huang BM, Liu MY. Parenteral sesame oil attenuates oxidative stress after endotoxin intoxication in rats. Toxicol 2004;196(1-2):147-53.
Bounous G, Molson JH. The antioxidant system. Anticancer Res 2003;23(2B):1411-5.
Balbis E, Patriarca S, Furfaro AL, Millanta S, Sukkar SG, Marinari UM, et al. Whey proteins influence hepatic glutathione after CCl4 intoxication. Toxicol Ind Health 2009;25(4-5):325-8.
Morr CV, Ha EY. Whey protein concentrates and isolates: processing and functional properties. Crit Rev Food Sci Nutr 1993;33(6):431-76.
Parmely MJ, Wang F, Wright D. Gamma interferon prevents the inhibitory effects of oxidative stress on host responses to Escherichia coli infection. Infect Immune 2001;69(4):2621-9.
Kim YM, de Vera ME, Watkins SC, Billiar TR. Nitric oxide protects cultured rat hepatocytes from tumor necrosis factor-alpha-induced apoptosis by inducing heat shock protein 70 expression. J Biol Chem 1997;272(2):1402-11.
Kudo H, Takahara T, Yata Y, Kawai K, Zhang W, Sugiyama T. Lipopolysaccharide triggered TNF-alpha-induced hepatocyte apoptosis in a murine non-alcoholic steatohepatitis model. J Hepatol 2009;51(1):168-75.
Kent KD, Harper WJ, Bomser JA. Effect of whey protein isolate on intracellular glutathione and oxidant-induced cell death in human prostate epithelial cells. Toxicol In vitro 2003;17(1):27-33.
Middleton N, Reid JR, Coolbear T, Jelen P. Proliferation and intracellular glutathione in Jurkat T cells with concentrated whey protein products. Int Dairy J 2003;13(7):565–73.
Rutherfurd-Markwick KJ, Johnson D, Cross ML, Gill HS. Modified milk powder supplemented with immunostimulating whey protein concentrate (IMUCARE) enhances immune function in mice. Nutr Res 2005;25:192-203.
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. Clin Diagn Lab Immunol 2001;8(6):1234-9.
Shimohama S. Apoptosis in Alzheimer's disease--an update. Apoptosis 2000;5(1):9-16.
Yang F, Sun X, Beech W, Teter B, Wu S, Sigel J, et al. Antibody to caspase-cleaved actin detects apoptosis in differentiated neuroblastoma and plaque-associated neurons and microglia in Alzheimer's disease. Am J Pathol 1998;152(2):379-89.
Wolfe JT, Ross D, Cohen GM. A role for metals and free radicals in the induction of apoptosis in thymocytes. FEBS Lett 1994;352(1):58-62.
Ramachandran A, Lebofsky M, Weinman SA, Jaeschke H. The impact of partial manganese superoxide dismutase (SOD2)-deficiency on mitochondrial oxidant stress, DNA fragmentation and liver injury during acetaminophen hepatotoxicity. Toxicol Appl Pharmacol 2011;251(3):226-33.
Abd EL-Aziem SH, Saleh ZA, Farrag AA. Impact of whey protein on the genotoxic effects of aflatoxins in rats. Int J Dairy Sci 2007;2 (2):126-37.
Peng X, Xiong YL, Kong B. Antioxidant activity of peptide fractions from whey protein hydrolysates as measured by electron spin resonance. Food Chem 2009;113:196-201.
Zhong J, Deaciuc IV, Burikhanov R, de Villiers WJ. Lipopolysaccharide-induced liver apoptosis is increased in interleukin-10 knockout mice. Biochim Biophys Acta 2006;1762(4):468-77.
Liu D, Li C, Chen Y, Burnett C, Liu XY, Downs S, et al. Nuclear import of proinflammatory transcription factors is required for massive liver apoptosis induced by bacterial lipopolysaccharide. J Biol Chem 2004;279(46):48434-42.
Canbay A, Feldstein AE, Higuchi H, Werneburg N, Grambihler A, Bronk SF, et al. Kupffer cell engulfment of apoptotic bodies stimulates death ligand and cytokine expression. Hepatol 2003;38(5):1188-98.
Canbay A, Friedman S, Gores GJ. Apoptosis: the nexus of liver injury and fibrosis. Hepatol 2004;39(2):273-8