Effects of N Acetylcysteine on Expression and Localization of Some Lectins in The Liver of Experimental Hepatic Intoxication

Öz

The aim of this study was to investigate the effects of intraperitoneal applications NAC that is a GSH precursor,determine the effects of some lectins expression and localization of the liver. Liver toxicity of CCl₄in rats in order to create, intraperitoneal (i.p.) 1 ml/kg 1/1 ratio of olive oil in the form of solution injected 3 times with an interval of one day. 24 hours after CCl₄injection, liver tissue taken under ether anesthesia. In order to determine the protective effect of N Acetyl Cysteine, N Acetyl Cysteine application (i.p. 50 mg/kg/day) 3 days prior to CCl₄injection started and continued during the experimental period. Animals liver tissue were taken 24. hours after the last injection under the ether anesthesia. Galactose (EEL), Lactose (R II) and Mannose (GNL) specific oligosaccharides were determined using biotinylated labeled lectins in liver tissue sections. Evaluation of the reaction was performed by light microscopy. Histochemically, the reactions have been observed in centrilobular vein sinusoidal capillaries of liver tissue. No staining was detected in hepatocytes. Compared with controls, CCl₄group were more intense staining. NAC reduced the staining intensity. As a result, cells increase their oligosaccharide units to protect them self from the liver damage of CCl₄. It was observed that NAC had protective and therapeutic effects against damage induced by CCl₄in rats hepatic cells.

Anahtar Kelimeler

• CCl4,EEL,GNL,NAC,Rat,RCA I

Deneysel Karaciğer İntoksikasyonunda N Asetil Sisteinin Karaciğerdeki Bazı Lektinlerin Ekspresyonu ve Lokalizasyonu Üzerine Etkileri

Öz

Yapılan çalışmada karaciğer dokusunda, GSH öncüsü NAS intraperitonal uygulamalarının, karaciğerdeki bazı lektinlerin ekspresyonu ve lokalizasyonu üzerine etkilerinin belirlenmesi amaçlanmıştır. Karaciğer toksisitesi oluşturmak amacıyla, sıçanlara CCl_4,periton içi (i.p.) 1 ml/kg 1/1 oranında, zeytinyağındaki çözeltisi şeklinde, birer gün ara ile 3 defa enjekte edildi. CCl₄enjeksiyonundan 24 saat sonra eter anestezisi altında, karaciğer dokusu alındı. N Asetil Sisteinin koruyucu etkisinin belirlenmesi amacıyla, NAS uygulaması (periton içi 50 mg/kg/gün) CCl₄enjeksiyonundan 3 gün önce başladı ve deney süresince devam etti. Hayvanların sonenjeksiyondan 24 saat sonra eter anestezisi altında karaciğer dokuları alındı. Karaciğer dokusundan alınan kesitlerde Galaktoza (EEL), Laktoza (RCA I) ve Mannoza (GNL)spesifik oligosakkarit üniteleribiotinlenmişişaretli lektinlerin kullanılması ile belirlendi.Reaksiyonların değerlendirilmesi ışık mikroskobunda gerçekleştirildi. Histokimyasal olarak reaksiyonlar; sentralobüler damar ve karaciğer dokusunun, sinüsoidal kapillerlerinde gözlemlendi. Hepatositlerde boyanma tespit edilmedi. Kontrollere göre CCl₄grubunda, daha yoğun bir boyama tespit edildi. NAS ise boyama yoğunluğunu azalttı. Sonuç olarak, hücrelerin CCl₄’e bağlı karaciğer hasarından kendilerini korumak için oligosakkarit ünitelerini artırdığı görüldü. NAS’ın, sıçanların hepatik hücrelerinde CCl₄ile oluşturulan hasara karşı koruyucu ve tedavi edici etkileri olduğu gözlemlendi.

Anahtar Kelimeler

• CCl4,EEL,GNL,NAS,Rat,RCA I

Kaynakça

1. Akşit H, Akşit D, Bildik A, Kara H, Yavuz Ö, Seyrek K (2015). Deneysel karaciğer intoksikasyonunda N-Asetil Sistein’in glutatyon metabolizması ve lipid peroksidasyonuna etkileri. Ankara Üniv Vet Fak Derg, 62, 1-5. Bacigalupo ML, Manzi M, Rabinovich GA, Troncoso MF (2013). Hierarchical and selective roles of galectins in hepatocarcinogenesis, liver fibrosis and inflammation of hepatocellular carcinoma. World J Gastroenterology, 19 (47), 8831-8849. Baenziger JU, Fiete D (1979). Structural determinants of Ricinus communis agglutinin and toxin specificity for oligosaccharides. J Biol Chem, 254, 9795–9799. Bampton JLM, Shirlaw PJ, Topley S, Weller P, Wilton JM (1991). Human junctional epithelium: Demonstration of a new marker, its growth in vitro and characterization by lectin reactivity and keratin expression. J Invest Dermatol, 96, 708-717. Basu D, Nair JV, Appukuttan PS (1987). Oligosaccharide structure determination of glycoconjugates using lectins. J Biosci, 11(1-4), 41-46. Bulgakow AA, Park KI, Choi KS, Lim HK, Cho M (2004). Purification and characterisation of a lectin isolated from the manila clam ruditapes philippinarum in krea. Fish Shellfish Immun, 16, 487-499. Even SWB, Pusztai A (1999). Effect of diets containing genetically modified potatoes expressing Galanthus Nivalis Lectin on rat small intestine. Lancet, 9187, 1353-1354. Franz H (1986). Mistletoe lectins and their a and b chains. Oncology, (43), 23-34. George S, Oh Y, Lindblom S, Vilain S, Rosa AJM, Françis DH, Brözel VS, Kaushik RS (2007). Lectin binding profile of the small intestine of five-week old pigs in response to the use of chlortetracycline as a growth promotant and under gnotobiotic conditions. J Anim Sci, 85, 1640-1650. Handa SS, Sharma A (1990). Hepatoprotective activity of andrographolide from Andrographis paniculata against carbontetrachloride. Indian J Med Res, 92, 276-283. Harrison FL (1991). Soluble vertebrate lectins: Ubiquitous but inscrutable proteins. J Cell Sci, 100, 9-14. Hormia M, Virtanen I (1989). Saccharide residues in human gingiva as revealed with fluorochrome-coupled lectins. J Periodont Res, 2, 137–145. Howard RJMW, Blake DR, Pall H, Williams A, Green ID (1987). Allopurinol/N-Acetylcysteine for carbon monoxide poisoning. Lancet, 2, 628-629. Kayalı H (1992). Özel Histoloji. Cerrahpaşa Tıp Fakültesi, İstanbul, Türkiye. Kelly GS (1998). Clinical applications of N-Acetylcysteine. Altern Med Rev, 3, 114-127. Kilpatrick DC (2002). Animal lectins: A historical introduction and overview. Biochim Biophys Acta, (2-3), 187-197. Kurşunlu SF (2011). Deneysel diyabet oluşturulan ratlarda dişeti dokusunun ekstrasellüler matriks ve hücre yüzeyinde bulunan glikokonjugatların yapısı ve lokalizasyonunun incelenmesi. Sağlık Bilimleri Enstitüsü, Biyokimya Anabilim Dalı, Doktora tezi, Aydın, Adnan Menderes Üniversitesi. Kus I, Ogeturk M, Oner H, Sahin S, Yekeler H, Sarsilmaz M (2005). Protective effects of melatonin against carbon tetrachloride-induced hepatotoxicity in rats: A light microscopic and biochemical study. Cell Biochem Funct, 3, 169-174. Liener I, Sharon N, Goldstein IJ (1986). The Lectins: Properties, Functions and Applications in Biology and Medicine, San Diego, Academic Press Inc. Lis H, Sharon N (1986). Lectins as molecules and as tools. Ann Rev Biochem, 55, 35-67. Mayanski DN, Schwartz YSH, Kutina SN, Zubakhin AA, Mayanskaya NN, Tsyrendorjiev DD (1993). Mononuclear phagocyte system responsiveness in CCl4-induced liver cirrhosis. Int J Exp Pathol, 74 (3), 229-236. Murnane RD, Ahern-Rindell AJ, Prieur DJ (1989). Lectin histochemistry of an ovine lysosomal storage disease with deficiencies of β-Galactosidase and α-Neuraminidase. Am J Pathol, 4, 135. Qua CS, Goh KL (2011). Liver cirrhosis in Malaysia: Peculiar epidemiology in a multiracial Asian country. J Gastroenterol Hepatol, 26, 1333-1337. Rojkind M (1973). Inhibition of liver fibrosis by-L Azetidine-2-carboxylic acid in rats treated with carbon tetrachloride. J Clin Invest, 52, 2451-2456. Scillitani GS, Zizza GE, Liquori GE, Ferri D (2007). Lectin histochemistry of gastrointestinal glycoconjugates in the greater horseshoe bat, Rhinolophus ferrumequinum. Acta Histochem, 109, 347-357. Sener G, Tosun O, Sehirli AO, Kacmaz A, Arbak S, Ersoy Y (2003). Melatonin and N-Acetylcysteine have beneficial effects during hepatic ischemia and reperfusion. Life Sci, 72, 2707-2718. Setshedi M, Longato L, Petersen DR, Ronis M, Chen WC, Wands JR, de la Monte SM (2011). Limited therapeutic effect of N-acetylcysteine on hepatic insulin resistance in an experimental model of alcohol-induced steatohepatitis. Alcohol Clin Exp Res, 35 (12), 2139-2151. Van Damme EJM, Allen AK, Peumans WJ (1987). İsolation and characterization of a lectin with exclusive specificity towards mannose from snowdrop (Galanthus nivalis) bulbs. FEBS Lett, 215, 140-144. Van Damme EJM, Peunians WJ (1988). Related mannose-specific lectins from different species of the family Amaryllidaceae. Plant Physiol Plant, 73, 52-57. Wieser R, Brunner G (1982). Interaktions- und Regulationsmechanismen der Zelle: Membranlektine- Membran glykomoleküle Biologie in unserer Zeit. 12, 97-107. Zaccone G, Fasula S, Locascio P, Licata A, Ainis L, Affronte R (1987). Lectin binding pattern on the surface epidermis of Ambystoma tigrinim larvae. Histochem, 87, 431-438. Zafarullah M, Li WQ, Sylvester J, Ahmad M (2003). Molecular mechanisms of N-Acetylcysteine actions. Cell Mol Life Sci, 60, 6-20.