Heparin modulation on hepatic nitric oxide synthase in experimental steatohepatitis

Hassanin,Amal; Malek,Hala Abdel; Saleh,Dalia

doi:10.3892/etm.2014.1963

Heparin modulation on hepatic nitric oxide synthase in experimental steatohepatitis

Authors:
- Amal Hassanin
- Hala Abdel Malek
- Dalia Saleh
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Affiliations: Department of Clinical Pharmacology, Faculty of Medicine, Mansoura University, Mansoura 35516, Egypt, Department of Anatomy and Embryology, Faculty of Medicine, Mansoura University, Mansoura 35516, Egypt
Published online on: September 15, 2014 https://doi.org/10.3892/etm.2014.1963
Pages: 1551-1558

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Abstract

Nonalcoholic fatty liver disease (NAFLD) is considered to be a hepatic manifestation of metabolic syndrome, and has been etiologically associated with insulin resistance (IR). The histopathology of NAFLD ranges between simple steatosis and nonalcoholic steatohepatitis (NASH), with or without fibrosis. The aim of the present study was to examine the effect of heparin on steatohepatitis and hepatic‑induced nitric oxide synthase (iNOS) expression in mice. Male mice were divided into four groups, which included the normal basal diet (control), high fat (HF) diet, HF diet + heparin (treatment group) and heparin control groups. After eight weeks from the initiation of the experiment, blood was collected and livers were harvested for biochemical analysis and histological studies. Serum levels of aspartate aminotransferase, alanine aminotransferase, hepatic triglyceride (TG) and hydroxyproline, as well as the IR, superoxide anion generation and mRNA expression of the hepatic iNOS enzyme were evaluated. Liver specimens were processed for histopathological and immunohistopathological evaluation. Heparin administration decreased the levels of the liver enzymes, IR, superoxide generation, hepatic TG, hydroxyproline and iNOS expression when compared with the HF diet group. These changes were associated with an improvement in inflammation and fibrosis observed via histopathological examination. Therefore, heparin treatment attenuates hepatic injury in steatohepatitis.

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1	Brunt EM, Neuschwander-Tetri BA, Oliver D, Wehmeier KR and Bacon BR: Nonalcoholic steatohepatitis: histologic features and clinical correlations with 30 blinded biopsy specimens. Hum Pathol. 35:1070–1082. 2004. View Article : Google Scholar : PubMed/NCBI
2	Marchesini G, Brizi M, Morselli-Labate AM, Bianchi G, Bugianesi E, McCullough AJ, Forlani G and Melchionda N: Association of nonalcoholic fatty liver disease with insulin resistance. Am J Med. 107:450–455. 1999. View Article : Google Scholar : PubMed/NCBI
3	Diehl AM: Lessons from animal models of NASH. Hepatol Res. 33:138–144. 2005. View Article : Google Scholar : PubMed/NCBI
4	Kita Y, Takamura T, Misu H, Ota T, Kurita S, Takeshita Y, Uno M, Matsuzawa-Nagata N, et al: Metformin prevents and reverses inflammation in a non-diabetic mouse model of nonalcoholic steatohepatitis. PLoS One. 7:e430562012. View Article : Google Scholar : PubMed/NCBI
5	McCullough AJ: Thiazolidinediones for nonalcoholic steatohepatitis - promising but not ready for prime time. N Engl J Med. 355:2361–2363. 2006. View Article : Google Scholar : PubMed/NCBI
6	Shah B and Shah G: Antifibrotic effect of heparin on liver fibrosis model in rats. World J Gastrointest Pharmacol Ther. 3:86–92. 2012.PubMed/NCBI
7	Howell M, Mohun TJ and Hill CS: Xenopus Smad3 is specifically expressed in the chordoneural hinge, notochord and in the endocardium of the developing heart. Mech Dev. 104:147–150. 2001. View Article : Google Scholar : PubMed/NCBI
8	Saï P, Pogu S and Ouary M: Heparin attenuates low-dose streptozotocin-induced immune diabetes in mice and inhibits the beta-cell binding of T-splenocytes in vitro. Diabetologia. 34:212–217. 1991.PubMed/NCBI
9	Lieber CS, Leo MA, Mak KM, Xu Y, Cao Q, Ren C, Ponomarenko A and Decarli LM: Model of nonalcoholic steatohepatitis. Am J Clin Nutr. 79:502–509. 2004.PubMed/NCBI
10	Bligh EG and Dyer WJ: A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 37:911–917. 1959. View Article : Google Scholar : PubMed/NCBI
11	Bergman M and Loxley R: The determination of hydroxyproline in urine hydrolysates. Clin Chim Acta. 27:347–349. 1970. View Article : Google Scholar : PubMed/NCBI
12	Wang HD, Pagano PJ, Du Y, Cayatte AJ, Quinn MT, Brecher P and Cohen RA: Superoxide anion from the adventitia of the rat thoracic aorta inactivates nitric oxide. Circ Res. 82:810–818. 1998. View Article : Google Scholar : PubMed/NCBI
13	Thiemermann C, Ruetten H, Wu CC and Vane JR: The multiple organ dysfunction syndrome caused by endotoxin in the rat: attenuation of liver dysfunction by inhibitors of nitric oxide synthase. Br J Pharmacol. 116:2845–2851. 1995. View Article : Google Scholar : PubMed/NCBI
14	Ekstedt M, Franzén LE, Mathiesen UL, Thorelius L, Holmqvist M, Bodemar G and Kechagias S: Long-term follow-up of patients with NAFLD and elevated liver enzymes. Hepatology. 44:865–873. 2006. View Article : Google Scholar : PubMed/NCBI
15	Chitturi S and Farrell GC: Etiopathogenesis of nonalcoholic steatohepatitis. Semin Liver Dis. 21:27–41. 2001. View Article : Google Scholar : PubMed/NCBI
16	Jansen PL: Non-alcoholic steatohepatitis. Eur J Gastroenterol Hepatol. 16:1079–1085. 2004. View Article : Google Scholar
17	McClain CJ, Mokshagundam SP, Barve SS, et al: Mechanisms of non-alcoholic steatohepatitis. Alcohol. 34:67–79. 2004. View Article : Google Scholar : PubMed/NCBI
18	Bonnefont-Rousselot D, Ratziu V, Giral P, Charlotte F, Beucler I and Poynard T; Lido Study Group. Blood oxidative stress markers are unreliable markers of hepatic steatosis. Aliment Pharmacol Ther. 23:91–98. 2006. View Article : Google Scholar : PubMed/NCBI
19	Albano E: Free radical mechanisms in immune reactions associated with alcoholic liver disease. Free Radic Biol Med. 32:110–114. 2002. View Article : Google Scholar : PubMed/NCBI
20	Kim F, Pham M, Maloney E, Rizzo NO, Morton GJ, Wisse BE, Kirk EA, Chait A and Schwartz MW: Vascular inflammation, insulin resistance, and reduced nitric oxide production precede the onset of peripheral insulin resistance. Arterioscler Thromb Vasc Biol. 28:1982–1988. 2008. View Article : Google Scholar : PubMed/NCBI
21	Guilherme A, Virbasius JV, Puri V and Czech MP: Adipocyte dysfunctions linking obesity to insulin resistance and type 2 diabetes. Nat Rev Mol Cell Biol. 9:367–377. 2008. View Article : Google Scholar : PubMed/NCBI
22	Poniachik J, Mancilla C, Contreras J, Csendes A, Smok G, Cavada G, Rojas J, et al: Obesity: risk factor for steatohepatitis and hepatic fibrosis. Rev Med Chil. 130:731–736. 2002.(In Spanish).
23	Larter CZ, Yeh MM, Haigh WG, Williams J, Brown S, Bell-Anderson KS, Lee SP and Farrell GC: Hepatic free fatty acids accumulate in experimental steatohepatitis: role of adaptive pathways. J Hepatol. 48:638–647. 2008. View Article : Google Scholar : PubMed/NCBI
24	Laurent A, Nicco C, Tran Van Nhieu J, Borderie D, Chéreau C, Conti F, et al: Pivotal role of superoxide anion and beneficial effect of antioxidant molecules in murine steatohepatitis. Hepatology. 39:1277–1285. 2004. View Article : Google Scholar : PubMed/NCBI
25	von Montfort C, Matias N, Fernandez A, Fucho R, Conde de la Rosa L, Martinez-Chantar ML, et al: Mitochondrial GSH determines the toxic or therapeutic potential of superoxide scavenging in steatohepatitis. J Hepatol. 57:852–859. 2012.PubMed/NCBI
26	Böhm T, Berger H, Nejabat M, Riegler T, Kellner F, Kuttke M, Sagmeister S, Bazanella M, et al: Food-derived peroxidized fatty acids may trigger hepatic inflammation: a novel hypothesis to explain steatohepatitis. J Hepatol. 59:563–570. 2013.PubMed/NCBI
27	Liu VW and Huang PL: Cardiovascular roles of nitric oxide: a review of insights from nitric oxide synthase gene disrupted mice. Cardiovasc Res. 77:19–29. 2008.PubMed/NCBI
28	Tateya S, Rizzo NO, Handa P, et al: Endothelial NO/cGMP/VASP signaling attenuates Kupffer cell activation and hepatic insulin resistance induced by high-fat feeding. Diabetes. 60:2792–2801. 2011. View Article : Google Scholar : PubMed/NCBI
29	Friedmann SL: Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem. 275:2247–2250. 2000. View Article : Google Scholar : PubMed/NCBI
30	Boisclair J, Doré M, Beauchamp G, Chouinard L and Girard C: Characterization of the inflammatory infiltrate in canine chronic hepatitis. Vet Pathol. 38:628–635. 2001. View Article : Google Scholar : PubMed/NCBI
31	Bataller R and Brenner D: Liver fibrosis. J Clin Invest. 115:209–218. 2005. View Article : Google Scholar
32	Mekonnen GA, Ijzer J and Nederbragt H: Tenascin-C in chronic canine hepatitis: immunohistochemical localization and correlation with necro-inflammatory activity, fibrotic stage, and expression of alpha-smooth muscle actin, cytokeratin 7, and CD3⁺ cells. Vet Pathol. 44:803–813. 2007. View Article : Google Scholar
33	Bedossa P and Paradis V: Liver extracellular matrix in health and disease. J Pathol. 200:504–515. 2003. View Article : Google Scholar : PubMed/NCBI
34	Guyot C, Lepreux S, Combe C, Doudnikoff E, Bioulac-Sage P, Balabaud C and Desmoulière A: Hepatic fibrosis and cirrhosis: the (myo)fibroblastic cell subpopulations involved. Int J Biochem Cell Biol. 38:135–151. 2006.PubMed/NCBI
35	Hinz B, Phan SH, Thannickal VJ, Galli A, Bochaton-Piallat M and Gabbiani G: The myofibroblast: one function, multiple origins. Am J Pathol. 170:1807–1816. 2007. View Article : Google Scholar : PubMed/NCBI
36	Ijzer J, Roskams T, Molenbeek RF, Ultee T, Penning LC, Rothuizen J and van den Ingh TS: Morphological characterization of portal myofibroblasts and hepatic stellate cells in the normal dog liver. Comp Hepatol. 5:72006. View Article : Google Scholar : PubMed/NCBI
37	Lever R and Page CP: Non-anticoagulant effects of heparin: an overview. Handb Exp Pharmacol. 207:281–305. 2012. View Article : Google Scholar : PubMed/NCBI
38	Mu E, Ding R, An X, Li X, Chen S and Ma X: Heparin attenuates lipopolysaccharide-induced acute lung injury by inhibiting nitric oxide synthase and TGF-β/Smad signaling pathway. Thromb Res. 129:479–485. 2012.
39	Horstman DJ, Fischer LG, Kouretas PC, Hannan RL and Rich GF: Role of nitric oxide in heparin-induced attenuation of hypoxic pulmonary vascular remodeling. J Appl Physiol (1985). 92:2012–2018. 2002.PubMed/NCBI
40	Ding R, Zhao D, Guo B, Zhang Z and Ma X: Treatment with unfractionated heparin attenuates coagulation and inflammation in endotoxemic mice. Thromb Res. 128:e160–e165. 2011. View Article : Google Scholar : PubMed/NCBI
41	Tokuyama Y, Nozaki O and Kanatsuka A: A patient with subcutaneous-insulin resistance treated by insulin lispro plus heparin. Diabetes Res Clin Pract. 54:209–212. 2001. View Article : Google Scholar : PubMed/NCBI
42	Fiorucci S, Antonelli E, Distrutti E, Severino B, Fiorentina R, Baldoni M, et al: PAR1 antagonism protects against experimental liver fibrosis. Role of proteinase receptors in stellate cell activation. Hepatology. 39:365–375. 2004. View Article : Google Scholar : PubMed/NCBI