Effect of palmitate-induced steatosis on paraoxonase-1 and paraoxonase-3 enzymes in human-derived liver (HepG2) cells

Year 2019, , 118 - 121, 01.12.2019

Gülben Sayılan Özgün , Eray Özgün Kıymet Tabakçıoğlu Selma Süer Gökmen Sevgi Eskiocak

https://doi.org/10.25000/acem.623975

Cited By: 1

Abstract

Aim: Palmitate is one of
the most abundant fatty acid in both liver of healthy individuals and in
patients with non-alcoholic fatty liver disease. Palmitate-induced
steatosis in HepG2 cells is an in vitro non-alcoholic fatty
liver disease model to investigate acute harmful effects of fat
overaccumulation in the liver. Non-alcoholic fatty liver disease is strongly
associated with atherosclerosis. Paraoxonase-1 and paraoxonase-3 are
anti-atherosclerotic enzymes which are bound to high density lipoprotein in
circulation and they are primarily synthesized by liver. There is no study that investigated the effect of palmitate-induced
steatosis on paraoxonase-1 and paraoxonase-3 enzymes. The aim of present
study was to investigate the effect of
palmitate-induced steatosis on paraoxonase-1 and paraoxonase-3 enzymes in HepG2 cells.

Methods: To induce steatosis, cells were incubated
with 0.4, 0.7 and 1 mM palmitate for 24 hours. Cell viability was
evaluated by 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium
bromide assay. Cells were stained with oil red O and triglyceride levels
were measured. Paraoxonase-1 and paraoxonase-3 protein levels were
measured by western blotting, their mRNA expression were measured by
quantitative PCR and arylesterase activity was measured spectrophotometrically.

Results: All palmitate
concentrations caused a significant increase on paraoxonase-1 mRNA levels.
Palmitate concentrations did not cause a significant change on paraoxonase-1 and paraoxonase-3 protein levels, paraoxonase-3 mRNA levels and
arylesterase activities.

Conclusion:
Our study showed that palmitate-induced steatosis
up-regulates paraoxonase-1
mRNA, has no effect on paraoxonase-1 and paraoxonase-3
protein levels, paraoxonase-3 mRNA
and arylesterase activity in HepG2 cells.

Keywords

Palmitate, paraoxonase-1, paraoxonase-3, arylesterase, HepG2, non-alcoholic fatty liver disease

Supporting Institution

No funding to declare.

Thanks

None

İnsan kaynaklı karaciğer (HepG2) hücrelerinde palmitat ile oluşturulan yağlanmanın paraoksonaz-1 ve paraoksonaz-3 enzimlerine etkisi

Abstract

Amaç: Palmitat, hem
sağlıklı bireylerin hem de non-alkolik karaciğer yağlanması hastalarının
karaciğerinde en fazla bulunan yağ asitlerinden biridir. HepG2 hücrelerinde
palmitat ile oluşturulan yağlanma, karaciğerdeki yağ birikiminin akut zararlı
etkilerinin araştırılmasında kullanılan in vitro non-alkolik yağlı karaciğer
hastalığı modelidir. Non-alkolik yağlı karaciğer hastalığı ateroskleroz ile
yakından ilişkilidir. Paraoksonaz-1 ve paraoksonaz-3 dolaşımda yüksek dansiteli
lipoproteine bağlı anti-aterosklerotik enzimlerdir ve esas olarak karaciğerde
sentezlenirler. Palmitat ile oluşturulan yağlanmanın paraoksonaz-1 ve
paraoksonaz-3 enzimleri üzerine etkisini araştıran bir çalışma bulunmamaktadır.
Bu çalışmanın amacı HepG2
hücrelerinde palmitat ile oluşturulan yağlanmanın paraoksonaz-1 ve
paraoksonaz-3 enzimlerine etkisini araştırmaktır.

Yöntemler: Yağlanma oluşturmak için hücreler 0.4, 0.7 ve
1 mM palmitat ile 24 saat inkübe edildi. Hücre canlılığı 3-(4,5-Dimetil-2-tiazolil)-2,5-difenil-2H-tetrazolium
bromür testi ile değerlendirildi. Hücreler oil red O ile boyandı ve trigliserit
düzeyleri ölçüldü. Paraoksonaz-1 ve paraoksonaz-3 protein düzeyleri western
blot ile, mRNA’ları ise kantitatif PCR ile ve arilesteraz aktivitesi
spektrofotometrik olarak ölçüldü.

Bulgular: Tüm palmitat konsantrasyonları
paraoksonaz-1 mRNA düzeylerinde anlamlı bir artışa yol açtı. Palmitat
konsantrasyonları paraoksonaz-1 ve paraoksonaz-3 protein düzeylerinde,
paraoksonaz-3 mRNA düzeylerinde ve arilesteraz aktivitesinde anlamlı bir
değişime yol açmadı.

Sonuç: Çalışmamız,
HepG2 hücrelerinde palmitat ile oluşturulan yağlanmanın paraoksonaz-1 mRNA
düzeyini arttırdığını, paraoksonaz-1 ve paraoksonaz-3
protein düzeylerine, paraoksonaz-3 mRNA düzeylerine ve arilesteraz aktivitesine
etkisi olmadığını gösterdi.

Keywords

Palmitat, paraoksonaz-1, paraoksonaz-3, arilesteraz, HepG2, non-alkolik yağlı karaciğer hastalığı

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