Lallemantia canescens (L) Fisch & Mey bitkisinin ve kallus doku kültürünün antioksidan aktivitesi

Abstract

Lallemantia canescens (L) Fisch & Mey Azerbeycan, Ermenistan, İran ve Türkiye’yi kapsayan geniş bir coğrafyada yetişmektedir. Çalışmamızda, Lallemantia canescens ve aynı bitkinin in vitro kallus kültürünün hem taze hem de metanol özütlerinin kararlı bir radikal olan 2,2-difenil-1- pikrilhidrazil (DPPH) ile reaktif oksijen türleri olan hidroksil (·OH), süperoksit (O2 .-) radikalleri ve lipid peroksidasyon inhibisyon özellikleri çalışılarak, antioksidan aktiviteleri araştırılmıştır. Deneylerde standart olarak bütillenmiş hidroksi toluen (BHT), kurkumin ve askorbik asit kullanılmıştır. Lallemantia canescens ile in vitro kallus kültürünün metanol özütlerinin toplam fenol, toplam flavonoid ve toplam antioksidan aktivitelerinin nicel olarak belirlenmesi için spektrofotometrik yöntemler kullanılmıştır. Özütlerin toplam fenol içerikleri Folin-Ciocalteu belirteciyle, toplam flavonoid miktarları aliminyum klorür metoduyla ve toplam antioksidan aktiviteleri ise fosfomolibden yöntemi ile tayin edilmiştir. Tüm Lallemantia canescens örneklerinin, kararlı bir radikal olan DPPH ile, Fe3-askorbat-EDTA – H2O2 sistemi ile oluşturulan hidroksil radikalini (·OH) temizledikleri saptanmıştır. Lallemantia canescens ve kallus doku kültürü metanol özütlerinin, ksantin-ksantin oksidaz sistemi ile oluşturulan süperoksit radikalini (O2 .-) ve sıçan karaciğer homojenatında tiyobarbütirik asit reaktif substratları (TBARS)’nın oluşumu ile belirlenen lipid peroksidasyonunu da inhibe ettikleri bulunmuştur. Bu sonuçlar doğrultusunda, Lallemantia canescens ile in vitro kallus kültürünün hem taze hemde metanol özütlerinin radikalleri temizleyerek ve lipid peroksidasyonunu inhibe ederek antioksidan aktiviteye sahip oldukları saptanmıştır. Lallemantia canescens ve kallus doku kültürü metanol özütlerinin yüksek toplam flavonoid ve toplam fenol içerikleri nedeniyle, biyoaktif moleküller için iyi birer kaynak olabilecekleri gözlenmiştir.

Lallemantia canescens (L) Fisch & Mey has spread out in wide location of Armenia, Azerbaijan, Iran and Turkey. The aim of the present work was to assay the antioxidant activities of fresh Lallemantia canescens and methanol extract, in addition in vitro callus tissue culture and callus tissue culture methanol extracts of Lallemantia canescens. BHT, curcumin and ascorbic acid was used as positive standards. It was determined that all the Lallemantia canescens examples scavenge hydroxyl radicals (·OH) generated by Fe3+-ascorbate-EDTA-H2O2 system. When the examples hydroxyl radical scavenging properties were examined based on IC50 which scavenging concentration %50 of the hydroxyl radical, it was found that decreases as plant methanol extract> fresh plant>callus tissue culture>callus tissue culture methanol extract respectively. It was seen that all examples had lower hydroxyl radical scavenging properties than the curcumin being used as a positive control. Whereas the other positive control BHT was found to have a lower hydroxyl radical scavenging properties than the fresh plant and plant methanol extract, but a higher scavenging property than the callus tissue culture and callus tissue culture methanol extract. Because the askorbic acid was in a experiment of hydroxyl radical environment, it was not used as a positive standard in hydroxyl radical scavenging inhibition When the DPPH radicals reducing properties were examined based on the IC50 values it was found that all the Lallemantia canescens examples reduced the stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH); plant methanol extract>fresh plant>callus tissue culture methanol extract>kallus tissue culture, respectively. It was seen that the ascorbic acid which of the positive control, has a higher DPPH radical reducing activity than all the examples. Curcumin had a higher than all examples except plant methanol extract whereas BHT had a lower than all examples except for the callus tissue culture, DPPH radical reducing properties. Plant methanol extract and callus tissue culture methanol extract have been found to both scavenged superoxide radicals (O2¯ ) generated by a xanthine and xanthine oxidase system and Fe(II) induced lipid peroxidation in rat liver homogenates, as indicated by decreased thiobarbituric acid-reactive substances (TBARS) formation. It was seen that the plant methanol extract was more active than the callus tissue culture methanol extract in superoxide radical scavenging and the lipid peroxidation inhibition. When the results were compared to positive controls it was found that curcumin and BHT are more active in superoxide scavenging and lipid peroxidation inhibiting than the plant methanol extract and callus tissue culture methanol extract samples. Because the askorbic acid was in a experiment lipid peroxidation environment, it was not used as a positive standard in lipid peroxidation inhibition but was found that its superoxide radical scavenging activity was lower than the other controls and extracts. In addition, spectrophotometric assay for the quantative determination of total antioxidant capacity, of plant methanol extract and callus tissue culture methanol extract was carried out. The extracts were analysed for the determination of total flavonoid by using the aluminium chloride method. Total phenol estimation was determined by the colorimetric method the Folin-Ciocaltes reagent. It was found that total phenolic, total flavonoid and total antioxidant activity properties were higher in plant methanol extract than the callus tissue culture methanol extract. Using these results it was determined that both fresh and methanol extracts of Lallemantia canescens in addition the in vitro callus tissue culture and callus tissue culture methanol extract has antioxidant activities by scavenging radicals and lipid peroxidation inhibition. It was observed that the Lallemantia canescens and the callus tissue culture methanol extracts could be a good source for bioactive molecules because of the high flavanoid and phenol quantities.