Kenevir tohumu yağının esansiyel yağ asitlerinin belirlenmesi ve akciğer kanser hücresi üzerinde in vitro olarak antikanserojenik özelliğinin araştırılması
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Dosyalar
Tarih
2022
Yazarlar
Dergi Başlığı
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Cilt Başlığı
Yayıncı
İnönü Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Kenevir (Cannabis), ana vatanı Orta Asya olarak bilinen ilk kültüre edilmiş bitkidir. Bitkinin tüm kısımları kullanım kolaylığı sağladığından endüstriyel bir bitki olarak tanınmaktadır. Kenevirin meyvesi olarak bilinen tohum kısmı ise "süper gıda" olarak kabul edilir. İçerisinde yüksek oranda yağ (% 35,5), karbonhidrat (% 27,6) ve protein (% 24,8) bulunmaktadır. Oranı % 35,5 olan yağlar çoğunlukla presleme yöntemiyle elde edilmektedir. Kenevir tohumu yağı içeriğinde yağ asitler (FA), esansiyel amino asitlerce zengin proteinler, karbonhidratlar, vitaminler (E, C, B1, B2, B3, B6) ve mineraller (P, K, Mg, S, Ca, Fe, Zn) bulunmaktadır. Kenevir tohumu yağı, esansiyel yağ asitleri ve amino asit içeriği bakımından çok değerlidir. Tohum yağındaki esansiyel yağ asitlerinden (EFA) ise çoklu doymamış yağ asitleri olarak bilinen linoleik asit (omega (ω) 3) ve α-linolenik asit (omega (ω) 6) ideal oranda (3:1) bulunmaktadır. Kenevir tohumu yağı içeriğindeki yağ asidi sayesinde, çeşitli metabolik hastalığın (kanser, nörolojik bozukluk, göz fonksiyonları, kardiyoloji vb.) tedavisi için olanak sağlamaktadır. Bu çalışmada, kenevir tohumundan soğuk presleme yöntemiyle kenevir tohumu yağı eldesi yapılmıştır. Elde edilen saf yağ, 10 ml n-hekzan ve 0.5 ml 2 N metanol içeren KOH çözeltisi ile muamele edilerek elde ettiğimiz metil ester türevli yağın GC-kromatogramı ile içeriğindeki esansiyel yağ asitleri tespit edilmiştir. Elimizde bulunan kenevir tohumu yağı örneğinin sonucunu "Supelco Fame mix 37" standardında belirtilen referans noktalı karşılaştırıldığında 19 adet yağ asidi belirlenmiştir. Analizde elde edilen sonuçlarda ise en yüksek yağ asitleri oranları sırasıyla; linoleik asit (LA) % 55,70, α-linolenik asit (ALA) % 18,19, oleik asit % 14,12, palmitik asit % 6,49 ve stearik asit % 2,79 olmuş diğer yağ asitler ise % 0,75'in altında kalmıştır. Bu yağ asitlerden LA ve ALA'nın hücre büyümesini düzenlediği, lipid peroksidasyonu, hücre proliferasyonu ve apoptoza sürükleme gibi antikanser etkileri olduğu ve oleik asidin ise kanser insidansını azaltmasıyla bilinmektedir. Ayrıca çalışmada, kenevir tohumu yağının akciğer kanser hücresi (A549) ve sağlıklı akciğer hücresi (BEAS-2B) üzerindeki etkisi araştırılmıştır. A549 ve BEAS-2B hücreleri kültür şişelerinde besiyer ortamında uygun yoğunluğa gelene kadar inkübatörde bekletildi. Kültür şişesindeki hücreler uygun yoğunluğuna ulaştıktan sonra 96 kuyucuklu plaklarda faklı yağ konsantrasyolarına (1 mg/ml, 5 mg/ml, 10 mg/ml, 25 mg/ ml, 50 mg/ml) maruz bırakılarak sonuçlar incelenmiştir. Plak okuyucuda okunan plaklar kontrol grupları ile karşılaştırmaları yapıldığında A549 ve BEAS-2B hücrelerinin proliferasyonunda ciddi bir düşüş görülmüştür. Bu düşün anlamlı bir düşüş olduğunu belirlemek için yapılan istatistik testi sonucunda A549 hücrelerine uygulanılan tüm konsantrasyonlarda p<0,05 olduğu saplanmış ve anlamlı olarak kabul edilmiştir. Ayrıca hücre proliferasyonundaki düşüşün apoptoz inüklenmesini analiz etmek için ise Flow sitometri (akan ölçer) yönteminde A549 hücre hattına 10 mg/ml ve 25 mg/ml yağ konsantrasyonu uygulandı ve hücre canlılığın oranın en düşük olduğu konsantrasyon 25 mg/ml (% 60,65) olmuştur. Analiz sonucunda akan sitometrik grafiği sonuçlarına bakıldığında kenevir tohumu yağı kanser hücrelerini apoptoza sürüklediği görülmüştür. Bu sonuçlara bakılarak kenevir tohumu yağının içeriğinde 19 esansiyel yağ asidi içeriği belirlenmiş ve kenevir tohumu yağının, akciğer kanser hücrelerinin hücre proliferasyonunu düşürdüğü ve hücre hatlarını apoptoza sürüklediği yani antikanserojenik etki yaptığı saptanmıştır.
Hemp (Cannabis) is the first cultivated plant, the homeland of which is known as Central Asia. It is recognized as an industrial plant, as all parts of the plant provide ease of use. The seed part, known as the fruit of hemp, is considered a "superfood". It has a high content of fats (35.5%), carbohydrates (27.6%) and proteins (24.8%). Oils with a ratio of 35.5% are most often obtained by pressing. The content of hemp seed oil includes fatty acids (FA), proteins rich in essential amino acids, carbohydrates, vitamins (E, C, B1, B2, B3, B6) and minerals (P, K, Mg, S, Ca, Fe, Zn). Hemp seed oil is very valuable for its content of essential fatty acids and amino acids. Seed oil, essential fatty acids (EFA) are known as polyunsaturated fatty acids, linoleic acid (omega (ω) 3) and α-linolenic acid (omega (ω) 6) the ratio (3:1) are there. Thanks to the fatty acid contained in hemp seed oil, it is possible to treat various metabolic diseases (cancer, neurological disorder, eye functions, cardiology, etc.) provides facilities for treatment. In this study, hemp seed oil was obtained from hemp seed by cold pressing method. The obtained pure oil was treated with 10 ml of n-hexane and 0.5 ml of 2 N methanol-containing KOH solution, and the essential fatty acids in its content were determined by GC-chromatogram of the methyl ester-derived oil we obtained. As a result of the hemp seed oil sample we have, 19 fatty acids have been determined compared to the reference points specified in the "Supelco Fame mix 37" standard. In the results obtained in the analysis, the highest ratios of fatty acids, respectively; linoleic acid (LA) was 55.70%, α-linolenic acid (ALA) was 18.19%, oleic acid was 14.12%, palmitic acid was 6.49% and stearic acid was 2.79%, while other fatty acids remained below 0.75%. Of these fatty acids, LA and ALA are known to regulate cell growth, have anticancer effects such as lipid peroxidation, cell proliferation, and drift to apoptosis, and oleic acid is known to reduce the incidence of cancer. In addition, the effect of hemp seed oil on lung cancer cells (A549) and healthy lung cells (BEAS-2B) was investigated in the study. A549 and BEAS-2B cells were kept in culture bottles in the incubator until they reached the appropriate density in the medium. After the cells in the culture bottle reached their proper density, the results were examined by exposing them to different oil concentrations (1 mg/ml, 5 mg/ml, 10 mg/ml, 25 mg/ml, 50 mg/ml) in 96-well plates. A significant decrease in the proliferation of A549 and BEAS-2B cells was observed when the plaques read in the plaque reader were compared with the control groups. As a result of the statistical test conducted to determine that there was a significant decrease in this value, it was found that p<0.05 was found at all concentrations applied to A549 cells and was considered significant. The decrease in cell proliferation, apoptosis inuklenme also for analyzing flow cytometry (flow meter) cell line A549 in the method 10 mg/mL and 25 mg/mL to the concentration of oil was applied, and the cell viability ratio is the lowest concentration of 25 mg/ml (% 60,65) has been. As a result of the analysis, when looking at the results of the flow cytometric graph, it was found that hemp seed oil leads cancer cells to apoptosis. Based on these results, the content of 19 essential fatty acids in the hemp seed oil was determined and it was found that hemp seed oil reduces the cell proliferation of lung cancer cells and causes cell lines to apoptosis, that is, has an anticarcinogenic effect.
Hemp (Cannabis) is the first cultivated plant, the homeland of which is known as Central Asia. It is recognized as an industrial plant, as all parts of the plant provide ease of use. The seed part, known as the fruit of hemp, is considered a "superfood". It has a high content of fats (35.5%), carbohydrates (27.6%) and proteins (24.8%). Oils with a ratio of 35.5% are most often obtained by pressing. The content of hemp seed oil includes fatty acids (FA), proteins rich in essential amino acids, carbohydrates, vitamins (E, C, B1, B2, B3, B6) and minerals (P, K, Mg, S, Ca, Fe, Zn). Hemp seed oil is very valuable for its content of essential fatty acids and amino acids. Seed oil, essential fatty acids (EFA) are known as polyunsaturated fatty acids, linoleic acid (omega (ω) 3) and α-linolenic acid (omega (ω) 6) the ratio (3:1) are there. Thanks to the fatty acid contained in hemp seed oil, it is possible to treat various metabolic diseases (cancer, neurological disorder, eye functions, cardiology, etc.) provides facilities for treatment. In this study, hemp seed oil was obtained from hemp seed by cold pressing method. The obtained pure oil was treated with 10 ml of n-hexane and 0.5 ml of 2 N methanol-containing KOH solution, and the essential fatty acids in its content were determined by GC-chromatogram of the methyl ester-derived oil we obtained. As a result of the hemp seed oil sample we have, 19 fatty acids have been determined compared to the reference points specified in the "Supelco Fame mix 37" standard. In the results obtained in the analysis, the highest ratios of fatty acids, respectively; linoleic acid (LA) was 55.70%, α-linolenic acid (ALA) was 18.19%, oleic acid was 14.12%, palmitic acid was 6.49% and stearic acid was 2.79%, while other fatty acids remained below 0.75%. Of these fatty acids, LA and ALA are known to regulate cell growth, have anticancer effects such as lipid peroxidation, cell proliferation, and drift to apoptosis, and oleic acid is known to reduce the incidence of cancer. In addition, the effect of hemp seed oil on lung cancer cells (A549) and healthy lung cells (BEAS-2B) was investigated in the study. A549 and BEAS-2B cells were kept in culture bottles in the incubator until they reached the appropriate density in the medium. After the cells in the culture bottle reached their proper density, the results were examined by exposing them to different oil concentrations (1 mg/ml, 5 mg/ml, 10 mg/ml, 25 mg/ml, 50 mg/ml) in 96-well plates. A significant decrease in the proliferation of A549 and BEAS-2B cells was observed when the plaques read in the plaque reader were compared with the control groups. As a result of the statistical test conducted to determine that there was a significant decrease in this value, it was found that p<0.05 was found at all concentrations applied to A549 cells and was considered significant. The decrease in cell proliferation, apoptosis inuklenme also for analyzing flow cytometry (flow meter) cell line A549 in the method 10 mg/mL and 25 mg/mL to the concentration of oil was applied, and the cell viability ratio is the lowest concentration of 25 mg/ml (% 60,65) has been. As a result of the analysis, when looking at the results of the flow cytometric graph, it was found that hemp seed oil leads cancer cells to apoptosis. Based on these results, the content of 19 essential fatty acids in the hemp seed oil was determined and it was found that hemp seed oil reduces the cell proliferation of lung cancer cells and causes cell lines to apoptosis, that is, has an anticarcinogenic effect.
Açıklama
Anahtar Kelimeler
Akciğer neoplazmları, Antikanser, Kenevir
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Yaman, Z. (2022). Kenevir tohumu yağının esansiyel yağ asitlerinin belirlenmesi ve akciğer kanser hücresi üzerinde in vitro olarak antikanserojenik özelliğinin araştırılması. Yayınlanmış Yüksek Lisans Tezi. İnönü Üniversitesi, Malatya.
