Öz
Aim: Exposure of the liver to diverse chemicals induce hepatic damage. Carbon tetrachloride (CCl4) is widely used toxin to investigate hepatic injury. In our study, the effects of naringin (NRG), a flavanone abundant in citrus fruits, on endoplasmic reticulum (ER) stress and stress-mediated apoptosis in CCl4- induced liver injury were investigated.
Materials and Methods: THLE-3 cells were exposed to varying concentrations of CCl4 for 24 hours and then treated with different doses of NRG for 4 hours. The effects of varying concentrations on cell viability were determined. Then, protein expressions of ER stress markers were detected in hepatocytes. Finally, Bcl2 active / inactive cell ratios were determined by flow cytometry.
Results: NRG treatment (5 and 10 µM) resulted in a significant increase in cell viability, which decreased with CCl4 administration. Similarly, the increased levels of ER stress markers as a result of
CCl4 application were significantly reduced with NRG treatment. Finally, NRG prevented apoptosis by significantly reducing ratio of Bcl2 inactive cells.
Conclusion: NRG treatment is effective in suppressing ER stress in CCl4-induced hepatocyte damage and preventing ER stress-induced apoptosis.
Anahtar Kelimeler
Liver damage naringin endoplasmic reticulum stress apoptosis
Kaynakça
- Novo E, Cannito S, Paternostro C, Bocca C, Miglietta A, Parola M. Cellular and molecular mechanisms in liver fibrogenesis. Arch Biochem Biophys. 2014; 548: 20-37.
- Tuñón MJ, Miguel BS, Crespo I, Jorquera F, Santamaría E, Alvarez M, et al. Melatonin attenuates apoptotic liver damage in fulminant hepatic failure induced by the rabbit hemorrhagic disease virus. J Pineal Res. 2011;50 (1): 38-45.
- Auzinger G, Wendon J. Intensive care management of acute liver failure. Current opinion in critical care. 2008;14(2):179-88.
- Yen F-L, Wu T-H, Lin L-T, Cham T-M, Lin C-C. Naringenin-loaded nanoparticles improve the physicochemical properties and the hepatoprotective effects of naringenin in orally-administered rats with CCl4-induced acute liver failure. Pharm Res. 2009; 26 (4): 893-902.
- Malhi H, Kaufman RJ. Endoplasmic reticulum stress in liver disease. J Hepatol. 2011; 54 (4): 795-809.
- Lin L, Que R, Shen Y, Chen Y, Yan N, Li Y. Saikosaponin‑d alleviates carbon‑tetrachloride induced acute hepatocellular injury by inhibiting oxidative stress and NLRP3 inflammasome activation in the HL‑7702 cell line. Molecular medicine reports. 2018; 17 (6): 7939-46.
- San‐Miguel B, Crespo I, Sánchez DI, González‐Fernández B, Ortiz de Urbina JJ, Tuñón MJ, et al. Melatonin inhibits autophagy and endoplasmic reticulum stress in mice with carbon tetrachloride‐induced fibrosis. J Pineal Res. 2015; 59 (2): 151-62.
- Hotamisligil GS. Endoplasmic reticulum stress and the inflammatory basis of metabolic disease. Cell. 2010; 140 (6): 900-17.
- Chen Q, Wu H, Tao J, Liu C, Deng Z, Liu Y, et al. Effect of naringin on gp120-induced injury mediated by P2X7 receptors in rat primary cultured microglia. PLoS One. 2017; 12 (8): e0183688.
- El-Desoky AH, Abdel-Rahman RF, Ahmed OK, El-Beltagi HS, Hattori M. Anti-inflammatory and antioxidant activities of naringin isolated from Carissa carandas L.: In vitro and in vivo evidence. Phytomedicine. 2018;42: 126-34.
- Shirani K, Yousefsani BS, Shirani M, Karimi G. Protective effects of naringin against drugs and chemical toxins induced hepatotoxicity: a review. Phytotherapy Research. 2020; 34 (8): 1734-44.
- Liu Y, Wu H, Nie Y-c, Chen J-l, Su W-w, Li P-b. Naringin attenuates acute lung injury in LPS-treated mice by inhibiting NF-κB pathway. International Immunopharmacology. 2011; 11 (10): 1606-12.
- Suzuki T, Motohashi H, Yamamoto M. Toward clinical application of the Keap1–Nrf2 pathway. Trends in pharmacological sciences. 2013; 34 (6): 340-6.
- Dong D, Xu L, Yin L, Qi Y, Peng J. Naringin prevents carbon tetrachloride-induced acute liver injury in mice. Journal of Functional Foods. 2015; 12: 179-91.
- Lu Q, Yang L, Zhao H-Y, Jiang J-G, Xu X-L. Protective effect of compounds from the flowers of Citrus aurantium L. var. amara Engl against carbon tetrachloride-induced hepatocyte injury. Food and chemical toxicology. 2013; 62: 432-5.
- Kim H-J, Song JY, Park HJ, Park H-K, Yun DH, Chung J-H. Naringin protects against rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells. The Korean Journal of Physiology & Pharmacology. 2009; 13 (4): 281-5.
- Stanca CM, Babar J, Singal V, Ozdenerol E, Odin JA. Pathogenic role of environmental toxins in immunemediated liver diseases. J Immunotoxicol. 2008; 5 (1): 59-68.
- Adams L, Angulo P. Treatment of non-alcoholic fatty liver disease. Postgrad Med J. 2006; 82 (967): 315-22.
- Ustuner D, Kolac UK, Ustuner MC, Tanrikut C, Ozdemir Koroglu Z, Burukoglu Donmez D, et al. Naringenin Ameliorate Carbon Tetrachloride-Induced Hepatic Damage Through Inhibition of Endoplasmic Reticulum Stress and Autophagy in Rats. J Med Food. 2020; 23 (11): 1192-200.
- Kim H-R, Lee G-H, Cho EY, Chae S-W, Ahn T, Chae H-J. Bax inhibitor 1 regulates ER-stress-induced ROS accumulation through the regulation of cytochrome P450 2E1. J Cell Sci. 2009; 122 (8): 1126-33.
- Wong FW-Y, Chan W-Y, Lee SS-T. Resistance to carbon tetrachloride-induced hepatotoxicity in mice which lack CYP2E1 expression. Toxicol Appl Pharmacol. 1998; 153 (1): 109-18.
- Lee G-H, Bhandary B, Lee E-M, Park J-K, Jeong K-S, Kim I-K, et al. The roles of ER stress and P450 2E1 in CCl4-induced steatosis. Int J Biochem Cell Biol. 2011; 43 (10): 1469-82.
- Kisseleva T, Brenner DA. Role of hepatic stellate cells in fibrogenesis and the reversal of fibrosis. Journal of gastroenterology and hepatology. 2007; 22: S73-S8.
- Atzori L, Poli G, Perra A. Hepatic stellate cell: a star cell in the liver. Int J Biochem Cell Biol. 2009; 41 (8-9): 1639-42.
- Bravo R, Parra V, Gatica D, Rodriguez AE, Torrealba N, Paredes F, et al. Endoplasmic reticulum and the unfolded protein response: dynamics and metabolic integration. International review of cell and molecular biology. 301: Elsevier; 2013. p. 215-90.
- Marumoto Y, Terai S, Urata Y, Matsumoto T, Mizunaga Y, Yamamoto N, et al. Continuous high expression of XBP1 and GRP78 is important for the survival of bone marrow cells in CCl4-treated cirrhotic liver. Biochem Biophys Res Commun. 2008; 367 (3): 546-52.
- Üstüner M, Tanrikut C, Üstüner D, Kolaç U, Köroğlu ZÖ, Burukoğlu D, et al. The effect of baicalein on endoplasmic reticulum stress and autophagy on liver damage. Human & Experimental Toxicology. 2021; 40 (10): 1624-33.
- Mukherjee PK. Quality control of herbal drugs: an approach to evaluation of botanicals: Business Horizons; 2002.
- Meng X, Tang G-Y, Liu P-H, Zhao C-J, Liu Q, Li H-B. Antioxidant activity and hepatoprotective effect of 10 medicinal herbs on CCl4-induced liver injury in mice. World J Gastroenterol. 2020; 26 (37): 5629.
- Chen L, Teng H, Zhang KY, Skalicka-Woźniak K, Georgiev MI, Xiao J. Agrimonolide and desmethylagrimonolide induced HO-1 expression in HepG2 cells through Nrf2-transduction and p38 inactivation. Frontiers in Pharmacology. 2017; 7: 513.
- Hu H, Tian M, Ding C, Yu S. The C/EBP homologous protein (CHOP) transcription factor functions in endoplasmic reticulum stress-induced apoptosis and microbial infection. Frontiers in immunology. 2019; 9: 3083.
- Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nature reviews Molecular cell biology. 2012; 13 (2): 89-102.
- Iurlaro R, Muñoz‐Pinedo C. Cell death induced by endoplasmic reticulum stress. The FEBS journal. 2016; 283 (14): 2640-52.
Öz
Amaç: Karaciğerin çeşitli kimyasallara maruz kalması hepatik hasar oluşmasına neden olur. Karbon tetraklorür (CCl4), kimyasal toksin kaynaklı hepatik hasarı araştırmak için yaygın şekilde
kullanılmaktadır. Çalışmamızda turunçgillerde bol bulunan bir flavanon olan naringinin (NRG), CCl4 ile indüklenen karaciğer hasarında endoplazmik retikulum (ER) stresi ve stres aracılı apoptoz üzerine etkileri incelenmiştir.
Gereç ve Yöntem: THLE-3 hücreleri 24 saat boyunca değişen konsantrasyonlarda CCl4’ e maruz bırakıldıktan sonra farklı dozlarda NRG ile 4 saat tedavi edildi. Konsantrasyonların hücre canlılığı
üzerindeki etkileri belirlendi. Ardından hepatositlerde ER stres belirteçlerinin protein ifadeleri tespit edildi. Son olarak akış sitometri ile Bcl2 aktif / inaktif hücre oranları belirlendi.
Bulgular: NRG tedavisi (5 ve 10 µM), CCl4 uygulaması ile azalan hücre canlılıklarında anlamlı bir artış sağladı. Benzer biçimde ER stres belirteçlerinde CCl4 uygulaması sonucu artan seviyeler NRG
tedavisiyle anlamlı biçimde azaldı. Son olarak NRG, Bcl2 inaktif hücre oranını anlamlı seviyede düşürerek apoptozun önüne geçilmesinde faydalı oldu.
Sonuç: NRG tedavisi CCl4 ile indüklenen hepatosit hasarında ER stresinin bastırılmasında ve ER stresi kaynaklı apoptozun önlenmesinde etkilidir.
Anahtar Kelimeler
Karaciğer hasarı naringin endoplazmik retikulum stresi apoptoz
Kaynakça
- Novo E, Cannito S, Paternostro C, Bocca C, Miglietta A, Parola M. Cellular and molecular mechanisms in liver fibrogenesis. Arch Biochem Biophys. 2014; 548: 20-37.
- Tuñón MJ, Miguel BS, Crespo I, Jorquera F, Santamaría E, Alvarez M, et al. Melatonin attenuates apoptotic liver damage in fulminant hepatic failure induced by the rabbit hemorrhagic disease virus. J Pineal Res. 2011;50 (1): 38-45.
- Auzinger G, Wendon J. Intensive care management of acute liver failure. Current opinion in critical care. 2008;14(2):179-88.
- Yen F-L, Wu T-H, Lin L-T, Cham T-M, Lin C-C. Naringenin-loaded nanoparticles improve the physicochemical properties and the hepatoprotective effects of naringenin in orally-administered rats with CCl4-induced acute liver failure. Pharm Res. 2009; 26 (4): 893-902.
- Malhi H, Kaufman RJ. Endoplasmic reticulum stress in liver disease. J Hepatol. 2011; 54 (4): 795-809.
- Lin L, Que R, Shen Y, Chen Y, Yan N, Li Y. Saikosaponin‑d alleviates carbon‑tetrachloride induced acute hepatocellular injury by inhibiting oxidative stress and NLRP3 inflammasome activation in the HL‑7702 cell line. Molecular medicine reports. 2018; 17 (6): 7939-46.
- San‐Miguel B, Crespo I, Sánchez DI, González‐Fernández B, Ortiz de Urbina JJ, Tuñón MJ, et al. Melatonin inhibits autophagy and endoplasmic reticulum stress in mice with carbon tetrachloride‐induced fibrosis. J Pineal Res. 2015; 59 (2): 151-62.
- Hotamisligil GS. Endoplasmic reticulum stress and the inflammatory basis of metabolic disease. Cell. 2010; 140 (6): 900-17.
- Chen Q, Wu H, Tao J, Liu C, Deng Z, Liu Y, et al. Effect of naringin on gp120-induced injury mediated by P2X7 receptors in rat primary cultured microglia. PLoS One. 2017; 12 (8): e0183688.
- El-Desoky AH, Abdel-Rahman RF, Ahmed OK, El-Beltagi HS, Hattori M. Anti-inflammatory and antioxidant activities of naringin isolated from Carissa carandas L.: In vitro and in vivo evidence. Phytomedicine. 2018;42: 126-34.
- Shirani K, Yousefsani BS, Shirani M, Karimi G. Protective effects of naringin against drugs and chemical toxins induced hepatotoxicity: a review. Phytotherapy Research. 2020; 34 (8): 1734-44.
- Liu Y, Wu H, Nie Y-c, Chen J-l, Su W-w, Li P-b. Naringin attenuates acute lung injury in LPS-treated mice by inhibiting NF-κB pathway. International Immunopharmacology. 2011; 11 (10): 1606-12.
- Suzuki T, Motohashi H, Yamamoto M. Toward clinical application of the Keap1–Nrf2 pathway. Trends in pharmacological sciences. 2013; 34 (6): 340-6.
- Dong D, Xu L, Yin L, Qi Y, Peng J. Naringin prevents carbon tetrachloride-induced acute liver injury in mice. Journal of Functional Foods. 2015; 12: 179-91.
- Lu Q, Yang L, Zhao H-Y, Jiang J-G, Xu X-L. Protective effect of compounds from the flowers of Citrus aurantium L. var. amara Engl against carbon tetrachloride-induced hepatocyte injury. Food and chemical toxicology. 2013; 62: 432-5.
- Kim H-J, Song JY, Park HJ, Park H-K, Yun DH, Chung J-H. Naringin protects against rotenone-induced apoptosis in human neuroblastoma SH-SY5Y cells. The Korean Journal of Physiology & Pharmacology. 2009; 13 (4): 281-5.
- Stanca CM, Babar J, Singal V, Ozdenerol E, Odin JA. Pathogenic role of environmental toxins in immunemediated liver diseases. J Immunotoxicol. 2008; 5 (1): 59-68.
- Adams L, Angulo P. Treatment of non-alcoholic fatty liver disease. Postgrad Med J. 2006; 82 (967): 315-22.
- Ustuner D, Kolac UK, Ustuner MC, Tanrikut C, Ozdemir Koroglu Z, Burukoglu Donmez D, et al. Naringenin Ameliorate Carbon Tetrachloride-Induced Hepatic Damage Through Inhibition of Endoplasmic Reticulum Stress and Autophagy in Rats. J Med Food. 2020; 23 (11): 1192-200.
- Kim H-R, Lee G-H, Cho EY, Chae S-W, Ahn T, Chae H-J. Bax inhibitor 1 regulates ER-stress-induced ROS accumulation through the regulation of cytochrome P450 2E1. J Cell Sci. 2009; 122 (8): 1126-33.
- Wong FW-Y, Chan W-Y, Lee SS-T. Resistance to carbon tetrachloride-induced hepatotoxicity in mice which lack CYP2E1 expression. Toxicol Appl Pharmacol. 1998; 153 (1): 109-18.
- Lee G-H, Bhandary B, Lee E-M, Park J-K, Jeong K-S, Kim I-K, et al. The roles of ER stress and P450 2E1 in CCl4-induced steatosis. Int J Biochem Cell Biol. 2011; 43 (10): 1469-82.
- Kisseleva T, Brenner DA. Role of hepatic stellate cells in fibrogenesis and the reversal of fibrosis. Journal of gastroenterology and hepatology. 2007; 22: S73-S8.
- Atzori L, Poli G, Perra A. Hepatic stellate cell: a star cell in the liver. Int J Biochem Cell Biol. 2009; 41 (8-9): 1639-42.
- Bravo R, Parra V, Gatica D, Rodriguez AE, Torrealba N, Paredes F, et al. Endoplasmic reticulum and the unfolded protein response: dynamics and metabolic integration. International review of cell and molecular biology. 301: Elsevier; 2013. p. 215-90.
- Marumoto Y, Terai S, Urata Y, Matsumoto T, Mizunaga Y, Yamamoto N, et al. Continuous high expression of XBP1 and GRP78 is important for the survival of bone marrow cells in CCl4-treated cirrhotic liver. Biochem Biophys Res Commun. 2008; 367 (3): 546-52.
- Üstüner M, Tanrikut C, Üstüner D, Kolaç U, Köroğlu ZÖ, Burukoğlu D, et al. The effect of baicalein on endoplasmic reticulum stress and autophagy on liver damage. Human & Experimental Toxicology. 2021; 40 (10): 1624-33.
- Mukherjee PK. Quality control of herbal drugs: an approach to evaluation of botanicals: Business Horizons; 2002.
- Meng X, Tang G-Y, Liu P-H, Zhao C-J, Liu Q, Li H-B. Antioxidant activity and hepatoprotective effect of 10 medicinal herbs on CCl4-induced liver injury in mice. World J Gastroenterol. 2020; 26 (37): 5629.
- Chen L, Teng H, Zhang KY, Skalicka-Woźniak K, Georgiev MI, Xiao J. Agrimonolide and desmethylagrimonolide induced HO-1 expression in HepG2 cells through Nrf2-transduction and p38 inactivation. Frontiers in Pharmacology. 2017; 7: 513.
- Hu H, Tian M, Ding C, Yu S. The C/EBP homologous protein (CHOP) transcription factor functions in endoplasmic reticulum stress-induced apoptosis and microbial infection. Frontiers in immunology. 2019; 9: 3083.
- Hetz C. The unfolded protein response: controlling cell fate decisions under ER stress and beyond. Nature reviews Molecular cell biology. 2012; 13 (2): 89-102.
- Iurlaro R, Muñoz‐Pinedo C. Cell death induced by endoplasmic reticulum stress. The FEBS journal. 2016; 283 (14): 2640-52.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Sağlık Kurumları Yönetimi |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 15 Mart 2023 |
| Gönderilme Tarihi | 21 Haziran 2022 |
| Yayımlandığı Sayı | Yıl 2023 |