Aspirinin insan kaynaklı hepatoma hücrelerinde paraoksonaz enzimlerinin protein düzeylerine ve arilesteraz aktivitesine etkisi
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.
Anahtar Kelimeler
Aspirin paraoksonaz-1 paraoksonaz-2 paraoksonaz-3 arilesteraz.
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
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
Anahtar Kelimeler
Aspirin paraoxonase-1 paraoxonase-2 paraoxonase-3 arylesterase.
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.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Konular | Sağlık Kurumları Yönetimi |
| Bölüm | Araştırma Makaleleri |
| Yazarlar | |
| Yayımlanma Tarihi | 20 Eylül 2019 |
| Gönderilme Tarihi | 11 Ekim 2018 |
| Yayımlandığı Sayı | Yıl 2019 |