Curcumin Regulated of Methotrexate-Induced Cell Damage in HEK-293 Cell Line

Öz

Objective: Methotrexate (MTX), a conventional anti-folate, is an anti-cancer agent that is widely used for treatment of various types of cancers and it has been researched due to its side effects. Unfortunately, toxic effects of MTX is not limited just with tumour cells. It has also a toxic effect on healthy cells and organs. That is why it is an indispensability that MTX should be combined with other agents which can reduce the cellular toxicity of it at the same time not reducing its anti-cancer effectiveness. Exhaustive investigations have been performed to reveal the usage of natural antioxidants that can be reduce the unwanted side effects of anti-cancer drugs. It is postulated that Curcumin (CUR) has a protective effect on cellular toxicity in various types of cells due to its antioxidant, anti-inflammatory and anti-cancer effects. This study was designed presuming that cellular toxicity induced by MTX can be reduced by Curcumin. In this study the regulatory role of curcumin, an antioxidant agent, was investigated on MTX treated Human Embryonic Kidney Cell lines (HEK-293). Materials and Methods: HEK-293 cells were divided into four groups as Control, CUR, MTX and MTX+CUR. Cells in the control group were incubated in cell culture medium for 48 hours without any application. After the cells in the other groups were incubated in cell culture medium for 24 hours cells in CUR group was treated with 10 μM CUR, cells in MTX group treated with 5 μM MTX and cells in the MTX+CUR group were incubated with 5 μM MTX + 10 μM CUR. After the treatments, the cells were kept incubated 24 hours in cell culture medium. Results: MTX-induced lipid peroxidation (Lip-Px) activity (according to method of Placer et al.,), glutathione (GSH- according to Sedlak and Lindsay method) and glutathione peroxidase (GSH-Px - according to the Lawrence and Burk method) levels were measured spectrophotometrically (UV-1800) in HEK-293 cells. It has been determined that Lip-Px activity increased and GSH and GSH-Px activities decreased significantly in cells incubated with MTX. It is established that CUR treatment caused a significant decrease on the Lip-Px activity while causing a significant increment on GSH and GSH-Px activities. Conclusion: Consequently, these findings showed that curcumin (CUR) administration, triggering the antioxidant mechanisms, can be an effective adjuvant therapy for MTX chemotherapy by regulating MTX-induced cellular stress and toxicity.

Anahtar Kelimeler

• Curcumin
• Glutathione peroxidase
• Methotrexate
• Lipid peroxidation.

Kurkumin HEK-293 Hücre Hattında Metotreksat Kaynaklı Hücresel Hasarı Düzenledi

Öz

Klasik bir antifolat olan metotreksat (MTX), çeşitli kanserlerin tedavisinde yaygın olarak kullanılan ve yan etkilerinden dolayı üzerinde çalışılan antikanser ajanlardan biridir. Ne yazık ki, MTX'in hücre üzerine toksik etkisi, sadece tümör hücreleri ile sınırlı olmayıp diğer hayati organları da etkilemektedir. Bu durum MTX’in antikanser etkinliğini azaltmadan, hücresel toksik etkilerini azaltabilecek başka ajanlarla birlikte kullanımını zorunlu kılmaktadır. Antikanser ilaçların istenmeyen yan etkilerini azaltabilecek doğal antioksidanların kullanımıyla ilgili kapsamlı araştırmalar yapılmaktadır. Yapılan çalışmalarda, kurkuminin (KUR) çeşitli dokularda meydana gelen hücresel toksisite üzerindeki koruyucu etkileri, onun antioksidan, antienflamatuar ve antikanser etkilerinin olmasına atfedilebilir. Bu çalışma, MTX’in neden olduğu hücresel toksisitenin, KUR ile azaltılabileceği varsayılarak yapıldı. MTX’e maruz bırakılan insan embriyo böbrek (HEK-293) hücre serisinde, antioksidan bir ajan olan KUR’un düzenleyici rolü araştırıldı. HEK-293 hücreleri, Kontrol, KUR, MTX ve MTX+KUR olarak dört gruba ayrıldı. Kontrol grubundaki hücrelere herhangi bir uygulama yapılmadan, kültür ortamında 48 saat boyunca tutuldu. Diğer gruplardaki hücreler kültür ortamında 24 saat tutulduktan sonra, KUR grubundaki hücrelere 10 μM KUR, MTX grubundaki hücrelere 5 μM MTX ve MTX+KUR grubundaki hücrelere ise 5 μM MTX ve 10 μM KUR uygulandı. Uygulamaları takiben hücreler 24 saat boyunca kültür ortamında tutuldu. HEK-293 hücrelerindeki MTX kaynaklı lipit peroksidasyon (Lip-Px) aktivitesi Placer ve arkadaşlarının yöntemine göre, glutatyon (GSH) seviyeleri Sedlak ve Lindsay yöntemine göre ve glutatyon peroksidaz (GSH-Px) seviyeleri Lawrence ve Burk yöntemine göre spektrofotometrik (UV-1800) olarak ölçüldü. MTX ile inkübe edilen hücrelerde Lip-Px aktivitesinin arttığı, GSH ve GSH-Px aktivitelerinin ise önemli ölçüde azaldığı belirlenmiştir. KUR uygulamasının ise Lip-Px aktivitesini önemli ölçüde azaltırken, GSH ve GSH-Px aktivitelerini önemli ölçüde artmıştır. Bu sonuçlar, KUR uygulamasının MTX kaynaklı hücresel stres ve toksisiteyi, antioksidan mekanizmalarla düzenleyerek, MTX kemoterapisine etkili bir yardımcı ajan olabileceğini göstermektedir.

Anahtar Kelimeler

• Kurkumin
• Metotreksat
• Lipit peroksidasyon

Kaynakça

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