Research Article
Hepatoprotective Effect Of Silymarin Against Diethylnitrosamine Induced Oxidative-Nitrosative Stress in Rats
Abstract
Liver cancer is the third most common cancer-related death in the World. Hepatocellular carcinoma (HCC) constitutes %70-85 of liver malignant primary tumors. In this study; we aimed to investigate the role of diethylnitrosamine (DEN) in the experimental liver cancer model and to examine the hepatoprotective effect of silymarin in detail using oxidative-nitrosative stress markers with immunohistochemical methods. For this purpose, 50 Wistar Albino male rats were divided into 5 groups with 10 rats. Animals in DEN group received DEN at a dose of 50 mg/kg once a week for 20 weeks. The rats in the DEN +Silymarin group were Silymarin intraperitoneally for 21 weeks 3 times a week at a dose of 100 mg/kg, starting 1 week prior to the administration of DEN. Silymarin was administered to rats in the group of Silymarin 3 times a week intraperitoneally for 20 weeks at a dose of 100 mg/kg. The rats in the control group were fed pellet feed and drinking water for 20 weeks. Animals in Sham group were treated with propylene glycol dissolved in 0,9 % NaCl intraperitoneally 3 times a week for 20 weeks. At the end of 21 weeks, macroscopically different size and number of nodular structures were detected in the DEN and DEN+Silymarin group. The microscopic examination revealed that the nodules were HCC. As a result, DEN induced liver cancer; however, silymarin did not show a statistically significant protective effect in terms of iNOS and Nitrotyrosine expressions.
Project Number
2016-TS-08
References
- Referans1Ahn B, Han BS, Kim DJ, Ohshima H (1999). Immunohistochemical localization of inducible nitric oxide synthase and 3-nitrotyrosine in rat liver tumors induced by N-nitrosodiethylamine. Carcinogenesis, 20 (7), 1337-1344.
- Referans2Ali F, Rahul, Naz F, Jyoti S, Siddique YH (2014). Protective effect of apigenin against N-nitrosodiethylamine (NDEA)-induced hepatotoxicity in albino rats. Mutat Res Genet Toxicol Environ Mutagen, 767, 13-20.
- Referans3Aparicio-Bautista DI, Pérez-Carreón JI, Gutiérrez-Nájera N, et al. (2013). Comparative proteomic analysis of thiol proteins in the liver after oxidative stress induced by diethylnitrosamine. Biochim Biophys Acta, 1834 (12), 2528-2538.
- Referans4Bishayee A, Barnes KF, Bhatia D, Darvesh AS, Carroll RT (2010). Resveratrol suppresses oxidative stress and inflammatory response in diethylnitrosamine-initiated rat hepatocarcinogenesis. Cancer Prev Res (Phila), 3 (6), 753-763.
- Referans5Bishayee A, Thoppil RJ, Darvesh AS, Ohanyan V, Meszaros JG, Bhatia D (2013). Pomegranate phytoconstituents blunt the inflammatory cascade in a chemically induced rodent model of hepatocellular carcinogenesis. J Nutr Biochem, 24 (1), 178-187.
- Referans6Cinel İ, Oral U (2000). Peroksinitrit ve Myokardium. Mersin Üniversitesi Tıp Fakültesi Dergisi 2, 163-168.
- Referans7Domitrović R, Jakovac H, Blagojević G (2011). Hepatoprotective activity of berberine is mediated by inhibition of TNF-α, COX-2, and iNOS expression in CCl(4)-intoxicated mice. Toxicology, 280 (1-2), 33-43.
- Referans8Ghosh D, Choudhury ST, Ghosh S, et al. (2012). Nanocapsulated curcumin: oral chemopreventive formulation against diethylnitrosamine induced hepatocellular carcinoma in rat. Chem Biol Interact, 195 (3), 206-214.
- Referans9Gupta P, Bansal MP, Koul A (2013). Lycopene modulates initiation of N-nitrosodiethylamine induced hepatocarcinogenesis: studies on chromosomal abnormalities, membrane fluidity and antioxidant defense system. Chem Biol Interact, 206 (2), 364-374.
- Referans10Kaviarasan S, Sundarapandiyan R, Anuradha CV (2008). Protective action of fenugreek (Trigonella foenum graecum) seed polyphenols against alcohol-induced protein and lipid damage in rat liver. Cell Biol Toxicol, 24 (5), 391-400.
- Referans11Kaya S, Eskiocak S, Tezel HA, et al. (2012). Kolorektal Kanserli Olgularda Oksidatif ve Nitrozatif Stres. Türk Klinik Biyokimya Derg 10 (2), 57-63.
- Referans12Kayalı R, Çakatay U (2004). Protein Oksidasyonunun Ana Mekanizmaları. Cerrahpaşa Tıp Dergisi 35 (2), 83-89
- Referans13Kim H, Hong MK, Choi H, Moon HS, Lee HJ (2015). Chemopreventive effects of korean red ginseng extract on rat hepatocarcinogenesis. J Cancer, 6 (1),1-8.
- Referans14Koç A (2011). Raw 264.7 Makrofaj Hücrelerinde Nitrik Oksit Sentezi Üzerine L-karnitinin Etkisi. Doktora Tezi. T.C. Ankara Üniversitesi Sağlık Bilimleri Enstitüsü, Ankara.
- Referans15Kumar AK, Vijayalakshmi K (2015). Protective effect of Punica granatum peel and Vitis vinifera seeds on DEN-induced oxidative stress and hepatocellular damage in rats. Appl Biochem Biotechnol, 175 (1), 410-420.
- Referans16Madrigal-Santillán E, Madrigal-Bujaidar E, Álvarez-González I, et al. (2014). Review of natural products with hepatoprotective effects. World J Gastroenterol, 20 (40),14787-14804.
- Referans17Mis L, Çilingir Yeltekin A (2014). Apoptosis. BEÜ Fen Bilimleri Dergisi, 3 (1), 102-112.
- Referans18Özen H, Kamber U, Karaman M, et al. (2014). Histopathologic, biochemical and genotoxic investigations on chronic sodium nitrite toxicity in mice. Exp Toxicol Pathol, 66 (8), 367-375.
- Referans19Özkan M, Yüksekol İ (2003). Nitrik Oksit ve Akciğerler. Toraks Dergisi, 4 (1), 88-94.
- Referans20Razavi-Azarkhiavi K, Ali-Omrani M, Solgi R, et al. (2014). Silymarin alleviates bleomycin-induced pulmonary toxicity and lipid peroxidation in mice. Pharm Biol, 52 (10),1267-1271.
- Referans21Sayılan Özgün G (2013). Streptozotosin ile Diyabet Geliştirilmiş Sıçanlarda Nitrik Oksit Metabolizmasının İncelenmesi. Doktora Tezi. T.C. Trakya Üniversitesi, Sağlık Bilimleri Enstitüsü Tıbbi Biyokimya Anabilim Dalı Doktora Programı, Edirne.
- Referans22Sayılan Özgün G, Özgün E, Eskiocak S, Süt N (2014). Streptozotosin ile diyabet oluşturulmuş sıçanlarda L-karnitinin nitric oksit metabolizması üzerine etkisi. Turk J Biochem, 39 (4), 416–421.
- Referans23Shaarawy SM, Tohamy AA, Elgendy SM, et al. (2009). Protective effects of garlic and silymarin on NDEA-induced rats hepatotoxicity. Int J Biol Sci, 5 (6), 549-557.
- Referans24Shirakami Y, Gottesman ME, Blaner WS (2012). Diethylnitrosamine-induced hepatocarcinogenesis is suppressed in lecithin:retinol acyltransferase-deficient mice primarily through retinoid actions immediately after carcinogen administration. Carcinogenesis, 33 (2), 268-274.
- Referans25Surai PF (2015). Silymarin as a Natural Antioxidant: An Overview of the Current Evidence and Perspectives. Antioxidants (Basel), 4 (1), 204-247.
- Referans26Thuy le TT, Morita T, Yoshida K, et al. (2011). Promotion of liver and lung tumorigenesis in DEN-treated cytoglobin-deficient mice. Am J Pathol, 179 (2), 1050-1060.
- Referans27Tunca R, Sozmen M, Citil M, Karapehlivan M, Erginsoy S, Yapar K (2009). Pyridine induction of cytochrome P450 1A1, iNOS and metallothionein in Syrian hamsters and protective effects of silymarin. Exp Toxicol Pathol, 61 (3), 243-255.
- Referans28Zhao X, Zhang JJ, Wang X, Bu XY, Lou YQ, Zhang GL (2008). Effect of berberine on hepatocyte proliferation, inducible nitric oxide synthase expression, cytochrome P450 2E1 and 1A2 activities in diethylnitrosamine- and phenobarbital-treated rats. Biomed Pharmacother, 62 (9), 567-572.
Ratlarda Dietilnitrozamin Uygulanarak Oluşturulan Oksidatif ve Nitrozatif Strese Karşı Silimarin’in Karaciğer Koruyucu Etkisi
Abstract
Karaciğer kanseri dünyada kanserle ilişkili ölümlerde üçüncü sırada yer almaktadır. Hepatoselüler karsinom (HSK) primer karaciğer kanserlerinin %70-85’ini oluşturmaktadır. Bu çalışmada; deneysel karaciğer kanseri modelinde Dietilnitrozamin (DEN)'in rolünü araştırmayı ve silimarinin hepatoprotektif etkisini oksidatif-nitrozatif stres markerlerı kullanarak immunohistokimyasal yöntemlerle ayrıntılı olarak incelemeyi amaçladık. Bu amaçla 50 adet Wistar Albino cinsi erkek rat 10’arlı 5 gruba ayrıldı. DEN grubundaki hayvanlara 50 mg/kg dozunda haftada 1 kez deneme süresince DEN uygulaması yapıldı. DEN+silimarin grubundaki ratlara DEN uygulamasından 1 hafta önce başlayacak şekilde 100 mg/kg dozunda haftada 3 kez deneme süresince intraperitoneal yolla silimarin (propilen glikolde çözdürülmüş) uygulandı. Kontrol grubundaki ratlara deneme süresince pelet yem ve içme suyu verildi. Sham grubundaki hayvanlara % 0.9 NaCl içerisinde çözdürülmüş propilen glikol intraperitoneal yolla haftada 3 kez deneme süresince verildi. Silimarin grubundaki ratlara 100 mg/kg dozunda deneme süresince intraperitoneal yolla haftada 3 kez silimarin uygulandı. 21 haftanın sonunda DEN ve DEN+silimarin gruplarında makroskobik olarak farklı büyüklük ve boyutlarda nodüler yapılar tespit edildi. Mikroskobik incelemede nodüllerin HSK oldukları belirlendi. Sonuç olarak DEN’in karaciğer kanserini indüklediği; silimarinin ise iNOS ve Nitrotirozin ekspresyonları açısından istatistiksel olarak anlamlı bir koruyucu etki göstermediği belirlenmiştir.
Supporting Institution
KAÜ Bilimsel Araştırma Projeleri Koordinatörlüğü
Project Number
2016-TS-08
Thanks
Bu çalışma KAÜ Bilimsel Araştırma Projeleri Koordinatörlüğü tarafından desteklenmiştir. Proje No: 2016-TS-08
References
- Referans1Ahn B, Han BS, Kim DJ, Ohshima H (1999). Immunohistochemical localization of inducible nitric oxide synthase and 3-nitrotyrosine in rat liver tumors induced by N-nitrosodiethylamine. Carcinogenesis, 20 (7), 1337-1344.
- Referans2Ali F, Rahul, Naz F, Jyoti S, Siddique YH (2014). Protective effect of apigenin against N-nitrosodiethylamine (NDEA)-induced hepatotoxicity in albino rats. Mutat Res Genet Toxicol Environ Mutagen, 767, 13-20.
- Referans3Aparicio-Bautista DI, Pérez-Carreón JI, Gutiérrez-Nájera N, et al. (2013). Comparative proteomic analysis of thiol proteins in the liver after oxidative stress induced by diethylnitrosamine. Biochim Biophys Acta, 1834 (12), 2528-2538.
- Referans4Bishayee A, Barnes KF, Bhatia D, Darvesh AS, Carroll RT (2010). Resveratrol suppresses oxidative stress and inflammatory response in diethylnitrosamine-initiated rat hepatocarcinogenesis. Cancer Prev Res (Phila), 3 (6), 753-763.
- Referans5Bishayee A, Thoppil RJ, Darvesh AS, Ohanyan V, Meszaros JG, Bhatia D (2013). Pomegranate phytoconstituents blunt the inflammatory cascade in a chemically induced rodent model of hepatocellular carcinogenesis. J Nutr Biochem, 24 (1), 178-187.
- Referans6Cinel İ, Oral U (2000). Peroksinitrit ve Myokardium. Mersin Üniversitesi Tıp Fakültesi Dergisi 2, 163-168.
- Referans7Domitrović R, Jakovac H, Blagojević G (2011). Hepatoprotective activity of berberine is mediated by inhibition of TNF-α, COX-2, and iNOS expression in CCl(4)-intoxicated mice. Toxicology, 280 (1-2), 33-43.
- Referans8Ghosh D, Choudhury ST, Ghosh S, et al. (2012). Nanocapsulated curcumin: oral chemopreventive formulation against diethylnitrosamine induced hepatocellular carcinoma in rat. Chem Biol Interact, 195 (3), 206-214.
- Referans9Gupta P, Bansal MP, Koul A (2013). Lycopene modulates initiation of N-nitrosodiethylamine induced hepatocarcinogenesis: studies on chromosomal abnormalities, membrane fluidity and antioxidant defense system. Chem Biol Interact, 206 (2), 364-374.
- Referans10Kaviarasan S, Sundarapandiyan R, Anuradha CV (2008). Protective action of fenugreek (Trigonella foenum graecum) seed polyphenols against alcohol-induced protein and lipid damage in rat liver. Cell Biol Toxicol, 24 (5), 391-400.
- Referans11Kaya S, Eskiocak S, Tezel HA, et al. (2012). Kolorektal Kanserli Olgularda Oksidatif ve Nitrozatif Stres. Türk Klinik Biyokimya Derg 10 (2), 57-63.
- Referans12Kayalı R, Çakatay U (2004). Protein Oksidasyonunun Ana Mekanizmaları. Cerrahpaşa Tıp Dergisi 35 (2), 83-89
- Referans13Kim H, Hong MK, Choi H, Moon HS, Lee HJ (2015). Chemopreventive effects of korean red ginseng extract on rat hepatocarcinogenesis. J Cancer, 6 (1),1-8.
- Referans14Koç A (2011). Raw 264.7 Makrofaj Hücrelerinde Nitrik Oksit Sentezi Üzerine L-karnitinin Etkisi. Doktora Tezi. T.C. Ankara Üniversitesi Sağlık Bilimleri Enstitüsü, Ankara.
- Referans15Kumar AK, Vijayalakshmi K (2015). Protective effect of Punica granatum peel and Vitis vinifera seeds on DEN-induced oxidative stress and hepatocellular damage in rats. Appl Biochem Biotechnol, 175 (1), 410-420.
- Referans16Madrigal-Santillán E, Madrigal-Bujaidar E, Álvarez-González I, et al. (2014). Review of natural products with hepatoprotective effects. World J Gastroenterol, 20 (40),14787-14804.
- Referans17Mis L, Çilingir Yeltekin A (2014). Apoptosis. BEÜ Fen Bilimleri Dergisi, 3 (1), 102-112.
- Referans18Özen H, Kamber U, Karaman M, et al. (2014). Histopathologic, biochemical and genotoxic investigations on chronic sodium nitrite toxicity in mice. Exp Toxicol Pathol, 66 (8), 367-375.
- Referans19Özkan M, Yüksekol İ (2003). Nitrik Oksit ve Akciğerler. Toraks Dergisi, 4 (1), 88-94.
- Referans20Razavi-Azarkhiavi K, Ali-Omrani M, Solgi R, et al. (2014). Silymarin alleviates bleomycin-induced pulmonary toxicity and lipid peroxidation in mice. Pharm Biol, 52 (10),1267-1271.
- Referans21Sayılan Özgün G (2013). Streptozotosin ile Diyabet Geliştirilmiş Sıçanlarda Nitrik Oksit Metabolizmasının İncelenmesi. Doktora Tezi. T.C. Trakya Üniversitesi, Sağlık Bilimleri Enstitüsü Tıbbi Biyokimya Anabilim Dalı Doktora Programı, Edirne.
- Referans22Sayılan Özgün G, Özgün E, Eskiocak S, Süt N (2014). Streptozotosin ile diyabet oluşturulmuş sıçanlarda L-karnitinin nitric oksit metabolizması üzerine etkisi. Turk J Biochem, 39 (4), 416–421.
- Referans23Shaarawy SM, Tohamy AA, Elgendy SM, et al. (2009). Protective effects of garlic and silymarin on NDEA-induced rats hepatotoxicity. Int J Biol Sci, 5 (6), 549-557.
- Referans24Shirakami Y, Gottesman ME, Blaner WS (2012). Diethylnitrosamine-induced hepatocarcinogenesis is suppressed in lecithin:retinol acyltransferase-deficient mice primarily through retinoid actions immediately after carcinogen administration. Carcinogenesis, 33 (2), 268-274.
- Referans25Surai PF (2015). Silymarin as a Natural Antioxidant: An Overview of the Current Evidence and Perspectives. Antioxidants (Basel), 4 (1), 204-247.
- Referans26Thuy le TT, Morita T, Yoshida K, et al. (2011). Promotion of liver and lung tumorigenesis in DEN-treated cytoglobin-deficient mice. Am J Pathol, 179 (2), 1050-1060.
- Referans27Tunca R, Sozmen M, Citil M, Karapehlivan M, Erginsoy S, Yapar K (2009). Pyridine induction of cytochrome P450 1A1, iNOS and metallothionein in Syrian hamsters and protective effects of silymarin. Exp Toxicol Pathol, 61 (3), 243-255.
- Referans28Zhao X, Zhang JJ, Wang X, Bu XY, Lou YQ, Zhang GL (2008). Effect of berberine on hepatocyte proliferation, inducible nitric oxide synthase expression, cytochrome P450 2E1 and 1A2 activities in diethylnitrosamine- and phenobarbital-treated rats. Biomed Pharmacother, 62 (9), 567-572.
There are 28 citations in total.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Journal Section | Araştırma Makaleleri |
| Authors | |
| Project Number | 2016-TS-08 |
| Publication Date | July 3, 2020 |
| Submission Date | January 27, 2020 |
| Acceptance Date | April 30, 2020 |
| Published in Issue | Year 2020 Volume: 31 Issue: 2 |
Cite
Accepted papers are licensed under Creative Commons Attribution-NonCommercial 4.0 International License