Hypericium perforatum extract (St. John’s Wort) and hypericin induce apoptosis in leukemic HL-60 cells by effecting h-TERT activity
Küçük Resim Yok
Tarih
2007
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Hiperisin, sarı kantaron (Hypericum perforatum L) olarak bilinen bitkinin aktif bileşenlerinden birisidir. Hi-perisinin antitümöral özellikleri olduğu solid organ tümörlerinin hücre dizileri ve hayvan modellerinde gösterilmiş ve bunun mitokondrial fonksiyonlarda bozulma neticesinde ortaya çıktığı gösterilmiştir. In vitro antilösemik etkisi de vardır. Ancak etki mekanizması tam olarak aydınlanabilmiş değildir. Hiperium ekstraktı ile hiperisine ait karşılaştırmalı sitotoksisite çalışması yoktur ve sitotoksisitenin altında yatan mekanizma aydınlatılamamış-tır. Bu çalışmada Ege Bölgesinde yetişen sarı kantaron otundan Hypericum perforatum ekstraktının elde edilip, bu ekstraktın HL-60 lösemik hücre dizisinde doz ve zamana bağımlı sitotoksisitesi olup olmadığını ve varsa bu sitotoksisik etkiyi hiperisinin sitotoksık özellikleri ile karşılaştırmak ve bu etkinin altında yatan mekanizmanın açıklanması amaçlanmıştır. Sarı kantaron otu ekstraktının 1/1000, 1/5000, 1/10.000, 1/50.000 dilüsyonları ile yapılan deneylerde, IC50 dozunun 1/10.000 dilüsyon olduğu görüldü. Hiperisin ile, lnM ve 100 ^M aralığında logaritmik artış gösteren dozlarda yapılan deneylerde de doz ve zamana bağımlı sitotoksisite olduğu ve 0.5 uJVI konsantrasyonunun, ekstraktın 1/10.000 dilüsyonunun gösterdiği eğriye paralel bir sitotoksisite gösterdiği tespit edildi. Hiperisinin sitotoksisite mekanizmasına yönelik olarak yapılan Acridine oranj/ethidium bromide boyamasında, 48. saatte belirgin olmak üzere IC50 dozunda, kontrollere oranla belirgin apoptozis varlığı saptanmıştır. Eş zamanlı olarak total RNA izole edilerek Light Cycler Real-time online polimeraz zincir reaksiyonu cihazında kit manueline uygun olarak h-TERT mRNA ekspresyonu çalışılmış ve 48. saatte belirgin ekspresyon azalması olduğu bulunmuştur. Sonuç olarak, kantaronun esas etken maddesi olan hiperisinin, HL-60 hücre dizisinde h-TERT ekspresyonunu etkileyerek apoptozisi uyardığı ortaya konmuştur.
Hypericin is the main active component of Hypericium perforatum (St. John's Wort). Hypericin has been proven to have antitumoral effect in in vitro condition against solid tumors by deteriorating the mitochondrial functions. It has also anti-leukemic effect in in vitro conditions. However, there has not been any comparative study with hypericin and extract obtained from Hypericium perforatum L. In this study, it has been aimed to investigate the potential cytotoxic role of the extract obtained from Hypericium perforatum grown in Ege region on leukemic cell line, to compare the cytotoxic effects of both extract and hypericin in HL-60 cells, and to clarify the underlying mechanism(s) of this cytotoxicity. Hypericium perforatum extract was used in dilutions as 1/1000, 1/5000, 1/10.000, 1/50.000 and the IC50 value was found to be as 1/10.000 dilution. Hypericin was found to have cytotoxicity in HL-60 cells in time and dose dependent manner between the doses of lnM to 100 M with ICgQ dose of 0.5 uJVI. Hypericin with the dose of 0.5 M had similar cytotoxicity pattern with the cytotoxicity curve obtained with 1/10000 diluted extract. Apoptosis as an underlying mechanism of this cytoxocity was shown in HL-60 cells after incubation with IC50 dose of hypericin which was more remarkable at 48th hours by using acridine orange/ethidium bromide dye method. Total RNA was isolated concomittantly and h-TERT mRNA expression was analyzed at Light Cycler Real-time online polymerase chain reaction and it was found that the mRNA expression was meaningfully decreased at 48th hour of incubation of cells with hypericin. According to-results of this study, we have shown that hypericin, as main cytotoxic compound of Hypericium perforatum I, induces apoptosis in HL-60 cells via effecting h-TERT mRNA expression
Hypericin is the main active component of Hypericium perforatum (St. John's Wort). Hypericin has been proven to have antitumoral effect in in vitro condition against solid tumors by deteriorating the mitochondrial functions. It has also anti-leukemic effect in in vitro conditions. However, there has not been any comparative study with hypericin and extract obtained from Hypericium perforatum L. In this study, it has been aimed to investigate the potential cytotoxic role of the extract obtained from Hypericium perforatum grown in Ege region on leukemic cell line, to compare the cytotoxic effects of both extract and hypericin in HL-60 cells, and to clarify the underlying mechanism(s) of this cytotoxicity. Hypericium perforatum extract was used in dilutions as 1/1000, 1/5000, 1/10.000, 1/50.000 and the IC50 value was found to be as 1/10.000 dilution. Hypericin was found to have cytotoxicity in HL-60 cells in time and dose dependent manner between the doses of lnM to 100 M with ICgQ dose of 0.5 uJVI. Hypericin with the dose of 0.5 M had similar cytotoxicity pattern with the cytotoxicity curve obtained with 1/10000 diluted extract. Apoptosis as an underlying mechanism of this cytoxocity was shown in HL-60 cells after incubation with IC50 dose of hypericin which was more remarkable at 48th hours by using acridine orange/ethidium bromide dye method. Total RNA was isolated concomittantly and h-TERT mRNA expression was analyzed at Light Cycler Real-time online polymerase chain reaction and it was found that the mRNA expression was meaningfully decreased at 48th hour of incubation of cells with hypericin. According to-results of this study, we have shown that hypericin, as main cytotoxic compound of Hypericium perforatum I, induces apoptosis in HL-60 cells via effecting h-TERT mRNA expression
Açıklama
Anahtar Kelimeler
Hematoloji
Kaynak
Turkish Journal of Hematology
WoS Q Değeri
Scopus Q Değeri
Cilt
24
Sayı
3
