Araştırma Makalesi
BibTex RIS Kaynak Göster

Aspirinin insan kaynaklı hepatoma hücrelerinde paraoksonaz enzimlerinin protein düzeylerine ve arilesteraz aktivitesine etkisi

Yıl 2019, , 295 - 302, 20.09.2019
https://doi.org/10.19161/etd.610836

Öz

Amaç: Bu çalışmanın amacı, aspirinin
insan kaynaklı hepatoma hücrelerinde paraoksonaz-1, paraoksonaz-2 ve paraoksonaz-3
protein düzeylerine ve arilesteraz aktivitesine etkisini araştırmaktır.

Gereç ve Yöntem: HepG2
hücreleri kontrol, 0.25 mM aspirin, 0.5 mM aspirin ve 1 mM aspirin olmak üzere
4 gruba ayrıldı ve hücreler 48 saat boyunca sırasıyla 0, 0.25, 0.5 ve 1 mM
aspirin ile inkübe edildi. Hücre canlılığı
3-(4,5-Dimetil-2-tiazolil)-2,5-difenil-2H-tetrazolium
bromür testi ile ölçüldü. Paraoksonaz-1, paraoksonaz-2 ve paraoksonaz-3 protein
düzeyleri western blot yöntemiyle ölçüldü. Arilesteraz aktivitesi substrat
olarak fenilasetat kullanılarak spektrofotometrik olarak ölçüldü.

Bulgular: 0.5 mM ve 1 mM aspirin
hücre canlılığında anlamlı azalmaya yol açtı. Aspirin konsantrasyonları
paraoksonaz-1 ve paraoksonaz-2 protein düzeylerini anlamlı olarak değiştirmedi.
0.5 ve 1 mM aspirin paraoksonaz-3 protein düzeylerini anlamlı olarak arttırdı.
0.25 mM, 0.5 mM ve 1 mM aspirin, arilesteraz aktivitesini anlamlı olarak
arttırdı.







Sonuç: Çalışmamız aspirinin
insan kaynaklı hepatoma hücrelerinde paraoksonaz-1 ve paraoksonaz-2 protein
düzeylerini değiştirmediğini, paraoksonaz-3 protein düzeylerini ve arilesteraz
aktivitesini arttırdığını gösterdi.

Kaynakça

  • Gaglia MA Jr, Clavijo L. Cardiovascular pharmacology core reviews: aspirin. J Cardiovasc Pharmacol Ther 2013; 18 (6): 505-13.
  • Schrör K, Rauch BH. Aspirin and lipid mediators in the cardiovascular system. Prostaglandins Other Lipid Mediat 2015; 121 (Pt A): 17-23.
  • She ZG, Chen HZ, Yan Y, Li H, Liu DP. The human paraoxonase gene cluster as a target in the treatment of atherosclerosis. Antioxid Redox Signal 2012; 16 (6): 597-632.
  • Chistiakov DA, Melnichenko AA, Orekhov AN, et al. Paraoxonase and atherosclerosis-related cardiovascular diseases. Biochimie 2017; 132: 19-27.
  • Précourt LP, Amre D, Denis MC, et al. The three-gene paraoxonase family: physiologic roles, actions and regulation. Atherosclerosis 2011; 214 (1): 20-36.
  • Ozgun E, Ozgun GS, Tabakcıoğlu K, Gokmen SS, Sut N, Eskıocak S. Effect of lipoic acid on paraoxonase-1 and paraoxonase-3 protein levels, mRNA expression and arylesterase activity in liver hepatoma cells. Gen Physiol Biophys 2017; 36 (4): 465-70.
  • Sayilan Ozgun G, Ozgun E, Tabakcıoğlu K, Suer Gokmen S, Eskiocak S, Cakir E. Caffeine increases apolipoprotein A-1 and paraoxonase-1 but not paraoxonase-3 protein levels in human-derived liver (HepG2) cells. Balkan Med J 2017; 34 (6): 534-9.
  • Draganov DI, Teiber JF, Speelman A, Osawa Y, Sunahara R, La Du BN. Human paraoxonases (PON1, PON2, and PON3) are lactonases with overlapping and distinct substrate specificities. J Lipid Res 2005; 46 (6): 1239-47.
  • Jaichander P, Selvarajan K, Garelnabi M, Parthasarathy S. Induction of paraoxonase 1 and apolipoprotein AI gene expression by aspirin. J Lipid Res 2008; 49 (10): 2142-48.
  • Santanam N, Parthasarathy S. Aspirin is a substrate for paraoxonase-like activity: implications in atherosclerosis. Atherosclerosis 2007; 191 (2): 272-75.
  • Bahar FG, Imai T. Aspirin hydrolysis in human and experimental animal plasmas and the effect of metal cations on their hydrolase activities. Drug Metab Dispos 2013; 41 (7): 1450-6.
  • Bouma ME, Rogier E, Verthier N, Labarre C, Feldmann G. Further cellular investigation of the human hepatoblastoma-derived cell line HepG2: morphology and immunocytochemical studies of hepatic-secreted proteins. In Vitro Cell Dev Biol 1989; 25 (3 Pt 1): 267-75.
  • Blatter-Garin MC, Kalix B, De Pree S, James RW. Aspirin use is associated with higher serum concentrations of the anti-oxidant enzyme, paraoxonase-1. Diabetologia 2003; 46 (4): 594-5.
  • Borthwick GM, Johnson AS, Partington M, Burn J, Wilson R, Arthur HM. Therapeutic levels of aspirin and salicylate directly inhibit a model of angiogenesis through a Cox-independent mechanism. FASEB J 2006; 20 (12): 2009-16.
  • Ahmadian S, Barar J, Saei AA, Fakhree, MA Omidi Y. Cellular toxicity of nanogenomedicine in MCF-7 cell line: MTT assay. J Vis Exp 2009; 26: 1191.
  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193: 265-75.
  • Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012; 9 (7): 671-5.
  • Gan KN, Smolen A, Eckerson HW, La Du BN. Purification of human serum paraoxonase/arylesterase. Evidence for one esterase catalyzing both activities. Drug Metab Dispos 1991;19(1):100-06.
  • Miao R, Xu X, Wang Z, Liu S, Qu K, Chen W, Liu C. Synergistic effect of nutlin-3 combined with aspirin in hepatocellular carcinoma HepG2 cells through activation of Bcl-2/Bax signaling pathway. Mol Med Rep 2018; 17 (3): 3735-43.
  • Liu YX, Feng JY, Sun MM, et al. Aspirin inhibits the proliferation of hepatoma cells through controlling GLUT1-mediated glucose metabolism. Acta Pharmacol Sin doi:10.1038/s41401-018-0014-x.
  • Kamble P, Selvarajan K, Aluganti Narasimhulu C, et al. Aspirin may promote mitochondrial biogenesis via the production of hydrogen peroxide and the induction of Sirtuin1/PGC-1α genes. Eur J Pharmacol. 2013 Jan 15; 699 (1-3): 55-61.
  • Aviram M, Rosenblat M, Bisgaier CL, et al. Paraoxonase inhibits high-density lipoprotein oxidation and preserves its functions. A possible peroxidative role for paraoxonase. J Clin Invest 1998;101(8):1581-90.
  • Shi X, Ding M, Dong Z, et al. Antioxidant properties of aspirin: characterization of the ability of aspirin to inhibit silica-induced lipid peroxidation, DNA damage, NF-κB activation, and TNF-α production. Mol Cell Biochem 1999; 199 (1-2): 93-102.
  • Ames PR, Batuca JR, Muncy IJ, et al. Aspirin insensitive thromboxane generation is associated with oxidative stress in type 2 diabetes mellitus. Thromb Res 2012; 130 (3): 350-4.
  • Kurban S, Mehmetoglu I. Effects of acetylsalicylic acid on serum paraoxonase activity, Ox-LDL, coenzyme Q10 and other oxidative stress markers in healthy volunteers. Clin Biochem. 2010; 43 (3): 287-90.
  • Draganov DI, Stetson PL, Watson CE, Billecke SS, La Du BN. Rabbit serum paraoxonase 3 (PON3) is a high density lipoprotein-associated lactonase and protects low density lipoprotein against oxidation. J Biol Chem 2000; 275 (43): 33435-42.

Effect of aspirin on protein levels of paraoxonase enzymes and arylesterase activity in human-derived hepatoma cells

Yıl 2019, , 295 - 302, 20.09.2019
https://doi.org/10.19161/etd.610836

Öz

Aim: The aim of this study is to investigate the effect of aspirin on
paraoxonase-1, paraoxonase-2 and paraoxonase-3 protein levels and arylesterase
activity in human-derived hepatoma cells.

Materials and Methods: HepG2 cells were divided into four groups: control,
0.25 mM aspirin, 0.5 mM aspirin and 1 mM aspirin and cells were incubated with
0, 0.25, 0.5 and 1 mM aspirin, respectively. Cell viability was evaluated by
3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay.
Paraoxonase-1, paraoxonase-2 and paraoxonase-3 protein levels were measured by
western blotting. Arylesterase activity was measured spectrophotometrically by
using phenylacetate as substrate.

Results: 0.5 mM and 1 mM aspirin caused a significant decrease on cell viability.
Aspirin concentrations did not significantly change paraoxonase-1 and
paraoxonase-2 protein levels. 0.5 mM and 1 mM aspirin significantly increased
paraoxonase-3 protein levels. 0.25 mM, 0.5 mM and 1 mM aspirin significantly
increased arylesterase activity.







Conclusion: Our study showed that
aspirin does not change paraoxonase-1 and paraoxonase-2 protein levels but increases
paraoxonase-3 protein levels and arylesterase activity in human-derived
hepatoma cells

Kaynakça

  • Gaglia MA Jr, Clavijo L. Cardiovascular pharmacology core reviews: aspirin. J Cardiovasc Pharmacol Ther 2013; 18 (6): 505-13.
  • Schrör K, Rauch BH. Aspirin and lipid mediators in the cardiovascular system. Prostaglandins Other Lipid Mediat 2015; 121 (Pt A): 17-23.
  • She ZG, Chen HZ, Yan Y, Li H, Liu DP. The human paraoxonase gene cluster as a target in the treatment of atherosclerosis. Antioxid Redox Signal 2012; 16 (6): 597-632.
  • Chistiakov DA, Melnichenko AA, Orekhov AN, et al. Paraoxonase and atherosclerosis-related cardiovascular diseases. Biochimie 2017; 132: 19-27.
  • Précourt LP, Amre D, Denis MC, et al. The three-gene paraoxonase family: physiologic roles, actions and regulation. Atherosclerosis 2011; 214 (1): 20-36.
  • Ozgun E, Ozgun GS, Tabakcıoğlu K, Gokmen SS, Sut N, Eskıocak S. Effect of lipoic acid on paraoxonase-1 and paraoxonase-3 protein levels, mRNA expression and arylesterase activity in liver hepatoma cells. Gen Physiol Biophys 2017; 36 (4): 465-70.
  • Sayilan Ozgun G, Ozgun E, Tabakcıoğlu K, Suer Gokmen S, Eskiocak S, Cakir E. Caffeine increases apolipoprotein A-1 and paraoxonase-1 but not paraoxonase-3 protein levels in human-derived liver (HepG2) cells. Balkan Med J 2017; 34 (6): 534-9.
  • Draganov DI, Teiber JF, Speelman A, Osawa Y, Sunahara R, La Du BN. Human paraoxonases (PON1, PON2, and PON3) are lactonases with overlapping and distinct substrate specificities. J Lipid Res 2005; 46 (6): 1239-47.
  • Jaichander P, Selvarajan K, Garelnabi M, Parthasarathy S. Induction of paraoxonase 1 and apolipoprotein AI gene expression by aspirin. J Lipid Res 2008; 49 (10): 2142-48.
  • Santanam N, Parthasarathy S. Aspirin is a substrate for paraoxonase-like activity: implications in atherosclerosis. Atherosclerosis 2007; 191 (2): 272-75.
  • Bahar FG, Imai T. Aspirin hydrolysis in human and experimental animal plasmas and the effect of metal cations on their hydrolase activities. Drug Metab Dispos 2013; 41 (7): 1450-6.
  • Bouma ME, Rogier E, Verthier N, Labarre C, Feldmann G. Further cellular investigation of the human hepatoblastoma-derived cell line HepG2: morphology and immunocytochemical studies of hepatic-secreted proteins. In Vitro Cell Dev Biol 1989; 25 (3 Pt 1): 267-75.
  • Blatter-Garin MC, Kalix B, De Pree S, James RW. Aspirin use is associated with higher serum concentrations of the anti-oxidant enzyme, paraoxonase-1. Diabetologia 2003; 46 (4): 594-5.
  • Borthwick GM, Johnson AS, Partington M, Burn J, Wilson R, Arthur HM. Therapeutic levels of aspirin and salicylate directly inhibit a model of angiogenesis through a Cox-independent mechanism. FASEB J 2006; 20 (12): 2009-16.
  • Ahmadian S, Barar J, Saei AA, Fakhree, MA Omidi Y. Cellular toxicity of nanogenomedicine in MCF-7 cell line: MTT assay. J Vis Exp 2009; 26: 1191.
  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193: 265-75.
  • Schneider CA, Rasband WS, Eliceiri KW. NIH Image to ImageJ: 25 years of image analysis. Nat Methods 2012; 9 (7): 671-5.
  • Gan KN, Smolen A, Eckerson HW, La Du BN. Purification of human serum paraoxonase/arylesterase. Evidence for one esterase catalyzing both activities. Drug Metab Dispos 1991;19(1):100-06.
  • Miao R, Xu X, Wang Z, Liu S, Qu K, Chen W, Liu C. Synergistic effect of nutlin-3 combined with aspirin in hepatocellular carcinoma HepG2 cells through activation of Bcl-2/Bax signaling pathway. Mol Med Rep 2018; 17 (3): 3735-43.
  • Liu YX, Feng JY, Sun MM, et al. Aspirin inhibits the proliferation of hepatoma cells through controlling GLUT1-mediated glucose metabolism. Acta Pharmacol Sin doi:10.1038/s41401-018-0014-x.
  • Kamble P, Selvarajan K, Aluganti Narasimhulu C, et al. Aspirin may promote mitochondrial biogenesis via the production of hydrogen peroxide and the induction of Sirtuin1/PGC-1α genes. Eur J Pharmacol. 2013 Jan 15; 699 (1-3): 55-61.
  • Aviram M, Rosenblat M, Bisgaier CL, et al. Paraoxonase inhibits high-density lipoprotein oxidation and preserves its functions. A possible peroxidative role for paraoxonase. J Clin Invest 1998;101(8):1581-90.
  • Shi X, Ding M, Dong Z, et al. Antioxidant properties of aspirin: characterization of the ability of aspirin to inhibit silica-induced lipid peroxidation, DNA damage, NF-κB activation, and TNF-α production. Mol Cell Biochem 1999; 199 (1-2): 93-102.
  • Ames PR, Batuca JR, Muncy IJ, et al. Aspirin insensitive thromboxane generation is associated with oxidative stress in type 2 diabetes mellitus. Thromb Res 2012; 130 (3): 350-4.
  • Kurban S, Mehmetoglu I. Effects of acetylsalicylic acid on serum paraoxonase activity, Ox-LDL, coenzyme Q10 and other oxidative stress markers in healthy volunteers. Clin Biochem. 2010; 43 (3): 287-90.
  • Draganov DI, Stetson PL, Watson CE, Billecke SS, La Du BN. Rabbit serum paraoxonase 3 (PON3) is a high density lipoprotein-associated lactonase and protects low density lipoprotein against oxidation. J Biol Chem 2000; 275 (43): 33435-42.
Toplam 26 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Sağlık Kurumları Yönetimi
Bölüm Araştırma Makaleleri
Yazarlar

Eray Özgün 0000-0002-6744-1519

Gülben Sayılan Özgün 0000-0001-6990-3484

Yayımlanma Tarihi 20 Eylül 2019
Gönderilme Tarihi 11 Ekim 2018
Yayımlandığı Sayı Yıl 2019

Kaynak Göster

Vancouver Özgün E, Sayılan Özgün G. Aspirinin insan kaynaklı hepatoma hücrelerinde paraoksonaz enzimlerinin protein düzeylerine ve arilesteraz aktivitesine etkisi. ETD. 2019;58(3):295-302.

1724617243172472652917240      26515    

 26507    26508 26517265142651826513

2652026519