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Investigaton of chemopreventive properties of Urtica dioica L., in MCF-7 and MDA231 breast cancer cell lines

Urtica dioica L., MCF-7 and MDA231 hücre hatları üzerindeki kemopreventif özelliklerinin araştırılması

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  1. Tez No: 305074
  2. Yazar: ELİF GÜLER
  3. Danışmanlar: PROF. DR. MESUDE İŞCAN
  4. Tez Türü: Doktora
  5. Konular: Biyoloji, Biology
  6. Anahtar Kelimeler: Neoplazmlar, Neoplasms
  7. Yıl: 2011
  8. Dil: İngilizce
  9. Üniversite: Orta Doğu Teknik Üniversitesi
  10. Enstitü: Fen Bilimleri Enstitüsü
  11. Ana Bilim Dalı: Biyoloji Bölümü
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: Belirtilmemiş.

Özet

Cancer is a major health problem in developing world with mostly unsufficient treatment. Cancer prevention through dietary modification appears to be a practical and cost effective possibility. The aim of present study is to investigate the chemical components of ?Urtica diocia,L (U. diocia) grown in Turkey? and the possible protective potential of its aqueous extract against breast cancer cell lines.U. diocia was extracted by maceration method which was performed for 6,12, 24, and 36 hours, at 50°C, 37°C, and 25°C for optimization of maceration condition. Total phenol and flavon contents, and radical scavenging activity (RSA) were determined. RSA was determined by DPPH (1,1diphenyl-2-picryl-hydrazil). RSA varied from %15 to %65 in different extracts prepared under different conditions. The highest RSA was found in 12 hour at 25°C extract with %65 RSA value and the (IC50) Inhibitory concentration of 0.30 mg/mL The extract obtained by maceration for 12h at 25°C had the richest flavon content 28.9 ±0.93 mg quercetin equivalent/g lyophilized extract. Phenol content of extracts in terms of gallic acid equivalent (GaE) varied from 28.8± 0.57 to 74.2 ± 2.24mg/g lyophilized extract. 12h at 25°C U. dioica extract had richest phenolic content (74.2± 2.24mg GaE /g lyophilized extract).The air dried aerial parts of the plant were extracted with methanol for 4 h at 400 C in sonicator. After evaporation of the combined methanol extracts of U.dioica under vacuum, the extract was fractionated with, chloroform, ethyl acetate and n-butanol by partitioning into solvents of increasing polarity. HPLC analyses of phenolic, compounds in n-butanol and ethyl acetate fractions of U.dioica were carried out in Varian ProStar HPLC. HPLC analyses of 12 h at 25°C U. dioica extract (14.4mg/mL) revealed that the extract contained 7.10 -3 mg/mL caffeic acid and 5.10-3 mg/mL quercetin. 3x 10-2mg/mL gallic acid was found in n-butanol fraction of U.dioica. Caffeic and chlorogenic acid were found at 1.9X 10-2 mg/mL in acid macerated extract of urtica dioica L.Cytotoxic effect of urtica dioica L in MCF-7 and MDA-231 cellls were investigated by using XTT Cell Proliferation assays which showed that urtica diocia L treatment for 48 hours caused a concentration dependent decrease in viable cell numbers. In vitro effects of U. diocia extracts on crude sheep liver cytosolic glutathione- S-transferase was also studied and have shown effective inhibition on cytosolic GST activity, with an IC50 of 75 nmoles/min/g protein.Total phenol and flavon content, and radical scavenging activity (RSA) were also determined to survey antioxidant potential of U. diocia acid extracts.Cancer is a major health problem in developing world with mostly unsufficient treatment. Cancer prevention through dietary modification appears to be a practical and cost effective possibility. The aim of present study is to investigate the chemical components of ?Urtica diocia,L (U. diocia) grown in Turkey? and the possible protective potential of its aqueous extract against breast cancer cell lines.U. diocia was extracted by maceration method which was performed for 6,12, 24, and 36 hours, at 50°C, 37°C, and 25°C for optimization of maceration condition. Total phenol and flavon contents, and radical scavenging activity (RSA) were determined. RSA was determined by DPPH (1,1diphenyl-2-picryl-hydrazil). RSA varied from %15 to %65 in different extracts prepared under different conditions. The highest RSA was found in 12 hour at 25°C extract with %65 RSA value and the (IC50) Inhibitory concentration of 0.30 mg/mL The extract obtained by maceration for 12h at 25°C had the richest flavon content 28.9 ±0.93 mg quercetin equivalent/g lyophilized extract. Phenol content of extracts in terms of gallic acid equivalent (GaE) varied from 28.8± 0.57 to 74.2 ± 2.24mg/g lyophilized extract. 12h at 25°C U. dioica extract had richest phenolic content (74.2± 2.24mg GaE /g lyophilized extract).The air dried aerial parts of the plant were extracted with methanol for 4 h at 400 C in sonicator. After evaporation of the combined methanol extracts of U.dioica under vacuum, the extract was fractionated with, chloroform, ethyl acetate and n-butanol by partitioning into solvents of increasing polarity. HPLC analyses of phenolic, compounds in n-butanol and ethyl acetate fractions of U.dioica were carried out in Varian ProStar HPLC. HPLC analyses of 12 h at 25°C U. dioica extract (14.4mg/mL) revealed that the extract contained 7.10 -3 mg/mL caffeic acid and 5.10-3 mg/mL quercetin. 3x 10-2mg/mL gallic acid was found in n-butanol fraction of U.dioica. Caffeic and chlorogenic acid were found at 1.9X 10-2 mg/mL in acid macerated extract of urtica dioica L.Cytotoxic effect of urtica dioica L in MCF-7 and MDA-231 cellls were investigated by using XTT Cell Proliferation assays which showed that urtica diocia L treatment for 48 hours caused a concentration dependent decrease in viable cell numbers. In vitro effects of U. diocia extracts on crude sheep liver cytosolic glutathione- S-transferase was also studied and have shown effective inhibition on cytosolic GST activity, with an IC50 of 75 nmoles/min/g protein.Total phenol and flavon content, and radical scavenging activity (RSA) were also determined to survey antioxidant potential of U. diocia acid extracts.Gelişmekte olan dünyanın en büyük sağlık problemi tedavide yetersiz kalınan kanser hastalığıdır. Kanserin önlenebilmesi beslenmede yapılacak düzenlemelerle uygulanabilir ve ekonomik açıdan mümkün olabilecektir. Çalışmanın amacı Türkiye'de yetişen ısırgan otunun var olabilecek kanser oluşumunu engelleyici özelliklerinin ve etki mekanizmalarının araştırılmasıdır.Bitki özütleri 6 ve 12, 24, 36. saatler ve 50°C, 37°C, 25°C sıcaklıkta maserasyon metoduyla hazırlanmıştır. Isırgan bitki özütünde, antioksidan aktivite tayin metodlarından toplam fenol, flavon ve serbest radikal süpürücü aktivitesi çalışılmıştır. Farklı koşullarda hazırlanan bitki özütlerinde yüzde radikal süpürücü etki (RSA) değerleri %15 ile %65 arasında değişmektedir. En yüksek RSA değeri 25°C'de, 12 saatte elde edilen bitki özütünde % 65 olarak bulundu. Bu özütün % 50 radikal süpürücü konsantrasyonu (IC50) 0.30 mg/mL olarak hesaplandı. En yüksek flavon içeriği 12 saat, 25 0C'de elde edilen örnekte 28.9±0.93 mg kuarsetin eşdeğeri/g kurutulmuş özüt olarak saptandı. Özütlerde fenol miktarının gallik asit eşdeğeri (GaE) olarak 28.8 ile 74.2 mg/g kurutulmuş özüt arasında değişmektedir. En yüksek fenol içeriği 12 saat, 25 0C'de hazırlanan özütte (74.2± 2.24 mg GaE/g kurutulmuş özüt) saptandı.U. dioica bitkisinde bulunan flavon ve fenolikler grubu aktif maddelerden kuarsetin ve kafeik asit miktarları; 12 saat, 25 0C de suyla hazırlanan özüt de, 7.10-3 mg/mL kafeik asit ve 5.10-3 mg/mL kuarsetin saptandı. 3x 10-2mg/mL Gallic acid n-butanol özütünde, 1.9X 10-2 mg/mL kafeik ve klorojenik asit, U. dioica bitkisinin asitle hazırlanan özütünde saptanmıştır.Isırgan otunun toprak üstü kısmından metanol ekstresi 4 saatte ve 40ºC'de sonikatör içerisinde hazırlanmıştır. Isırgan otu iki defa methanolde ekstre edilerek ve polarite farkından yararlanarak kloroform, etil asetat, n-butanol içerisinde fraksiyonlarına ayrılmıştır. Etilasetat ve n-butanol fraksiyonlarında içerik analizi, Varian Prostar HPLC kullanılarak yapılmıştır.U. dioica özütünün, 48 saat süreyle, artan konsantrasyonlarda MCF7 ve MDA-231 hücre kültürleri üzerindeki sitotoksik etkisi XTT yöntemi ile tespit edildi. Isırgan sulu özütün koyun karaciğer sitozolünde in vitro olarak GST enzim aktivitelerine bakıldı enzim aktivite inhibisyonu IC50 of 75 nmoles/min/g protein değeriyle ifade edildi.Asidik özütlerin total fenol, total flavon miktarları ve radikal süpürücü etkileri (RSA) tayin edilerek sulu özüt ile karşılaştırıldı.

Özet (Çeviri)

Cancer is a major health problem in developing world with mostly unsufficient treatment. Cancer prevention through dietary modification appears to be a practical and cost effective possibility. The aim of present study is to investigate the chemical components of ?Urtica diocia,L (U. diocia) grown in Turkey? and the possible protective potential of its aqueous extract against breast cancer cell lines.U. diocia was extracted by maceration method which was performed for 6,12, 24, and 36 hours, at 50°C, 37°C, and 25°C for optimization of maceration condition. Total phenol and flavon contents, and radical scavenging activity (RSA) were determined. RSA was determined by DPPH (1,1diphenyl-2-picryl-hydrazil). RSA varied from %15 to %65 in different extracts prepared under different conditions. The highest RSA was found in 12 hour at 25°C extract with %65 RSA value and the (IC50) Inhibitory concentration of 0.30 mg/mL The extract obtained by maceration for 12h at 25°C had the richest flavon content 28.9 ±0.93 mg quercetin equivalent/g lyophilized extract. Phenol content of extracts in terms of gallic acid equivalent (GaE) varied from 28.8± 0.57 to 74.2 ± 2.24mg/g lyophilized extract. 12h at 25°C U. dioica extract had richest phenolic content (74.2± 2.24mg GaE /g lyophilized extract).The air dried aerial parts of the plant were extracted with methanol for 4 h at 400 C in sonicator. After evaporation of the combined methanol extracts of U.dioica under vacuum, the extract was fractionated with, chloroform, ethyl acetate and n-butanol by partitioning into solvents of increasing polarity. HPLC analyses of phenolic, compounds in n-butanol and ethyl acetate fractions of U.dioica were carried out in Varian ProStar HPLC. HPLC analyses of 12 h at 25°C U. dioica extract (14.4mg/mL) revealed that the extract contained 7.10 -3 mg/mL caffeic acid and 5.10-3 mg/mL quercetin. 3x 10-2mg/mL gallic acid was found in n-butanol fraction of U.dioica. Caffeic and chlorogenic acid were found at 1.9X 10-2 mg/mL in acid macerated extract of urtica dioica L.Cytotoxic effect of urtica dioica L in MCF-7 and MDA-231 cellls were investigated by using XTT Cell Proliferation assays which showed that urtica diocia L treatment for 48 hours caused a concentration dependent decrease in viable cell numbers. In vitro effects of U. diocia extracts on crude sheep liver cytosolic glutathione- S-transferase was also studied and have shown effective inhibition on cytosolic GST activity, with an IC50 of 75 nmoles/min/g protein.Total phenol and flavon content, and radical scavenging activity (RSA) were also determined to survey antioxidant potential of U. diocia acid extracts.

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