BİR İLAÇ OLAN ANKAFERD BLOOD STOPPER'IN İNSAN UMBİLİKAL VEN ENDOTEL HÜCRE KÜLTÜRÜNE ETKİSİ The Effect of Ankaferd Blood Stopper as a Drug On Human Umbilical Vein Endothelial Cell Culture(HUVEC)
Abstract
ÖZET
Giriş: Ankaferd kanama durdurucu (ABS) tıbbi bitki ekstraksiyonudur. ABS Thymus vulgaris, Glicirhiza glabra,
Vitis vinifera, Alpinia officinarum ve Urtica dioica bitkilerinin standardize edilmiş bir karışımıdır. Bu çalışmanın
amacı in vitro endotel hücre çoğalmasının ABS'nin farklı zaman ve dozlarının in vitro apoptotik etkilerini
araştırmaktır.
Gereç ve yöntem: Erciyes Üniversitesi Hastanesinde sezeryanla yapılan doğumlardan alınan göbek bağı toplardamarı
PBS ile yıkandıktan sonra içine yaklaşık 10 mg/ml kollajenaz verildi ve 10 dakika 37ºC de inkübatörde
bekletildi. Damar içerisindeki hücre ve kollajenaz karışımı tüp içine alındıktan ve 30 ml besi yeri ilave
edildikten sonra 1000 rpm’de 10 dakika santrifüj edildi. Üstteki supernatant atıldı ve tüpün dip kısmında
biriken hücreler üzerine 2 ml besi yeri ilave edilerek karıştırıldı ve 25 cm’lik flasklara ekildi. Thoma lamında
sayılan hücreler gruplara ayrılarak her bir gruba ait kültür ortamına sırasıyla 5 µl, 25 µl and 50 µl ABS 24 saat
uygulandı. Daha sonra hücrelerin canlılığı ve/veya proliferasyonu MTS testi ile tespit edildi.
Bulgular: Doza bağlı olarak deney grubu hücrelerinde bir artışın olduğu görüldü. Kontrol ve deney gruplarından
elde edilen ortalama hücre sayısı sırasıyla 7,68x103 (±1,7), 5,56x103(±1,09) 4,12x103(±1,14) ve
2,43x103(±0,89) idi. Deney gruplarının kontrol grubu ile karşılaştırıldığında deney gruplarındaki azalma istatistiksel
olarak anlamlıydı (p<0,05). Deney grupları karşılaştırıldığında yüksek dozda hücre sayısı azdı ve
istatistiksel olarak anlamlı bulundu (p <0.05). Ayrıca, regresyon analizinin sonuçları hücre canlılığının doza
bağlı olarak azaldığını, yani dozun artmasıyla hücre ölümünün arttığını gösterdi.
Sonuç: Çalışmamızda ABS'nin apoptotik etkisi HUVEC'de gösterildi. Apoptoz, bir hücrede gerçekleşen biyokimyasal,
morfolojik ve moleküler değişikliklerden yararlanılarak çeşitli yöntemler aracılığı ile ölçülebilir.
ABS'nin apoptotik etkisi, kanser hastalıklarının tedavisinde alternatif bir yol olabileceğini göstermektedir.
Anahtar sözcükler: Ankaferd; Endotel; Umbilikal ven
ABSTRACT
Introduction: Ankaferd Blood Stopper (ABS) is an herbal extract. ABS comprises of standardized mixture of
herbs T. vulgaris, G. glabra, V. vinifera, A. officinarum and U. dioica. The aim of this study was to investigate
the in vitro apoptotic effects of the different times and doses of ABS on in vitro endothelial cell proliferation.
Methods: Umbilical cord obtained at Caesarean sections from Erciyes University hospital. After washing
PBS, the cord vein lumen was filled with PBS containing 10 mg/ml collagenase and incubated 10 minute
at 37°C. The contents of the vein were flushed out with 30 ml medium collected in centrifuge tube then
centrifugated at 1000 rpm for 10 minute. The cells were plated in 2 ml of medium at T-25 plastic flasks. The
endothelial cells were incubated with only medium in control group and different concentrations of ABS (5
µl, 25 µl and 50 µl ABS) in the experimental groups for 24 h. Then the viability and/or proliferation of cells
were detected by MTS assay.
Results: The mean cell number were obtained from the control and experimental groups 7,68x103 (±1,7),
5,56x103(±1,09) 4,12x103(±1,14), and 2,43x103(±0,89) respectively. The cells of the experimental groups
compared with the control group to the decrease in experimental groups was statistically significant
(p<0,05). Experimental groups were showed a decreased cell number in HUVEC compared with the each
other depended on high dose and was found to be statistically significant (p<0.05). Furthermore, the results
of regression analysis showed that cell viability was decreased depending on the dose, that was, the cell
death was increased as the dose was increased.
Conclusion: In our study, the apoptotic effect of ABS was investigated in HUVEC. The apoptosis can be
measured using various of methods by taking advantage of the biochemical, morphological, and molecular
changes undergoing in a cell during this process. The apoptotic effect of ABS shows that it may be an alternative
way of treating cancer diseases.
Key words: Ankaferd; Endothelial; Umbilical vein
References
- 1. Dinçol ME, Ozbas H, Yılmaz B, Ersev H, Gokyay S and Olgac V. Effect of the plant-based hemostatic agent Ankaferd Blood Stopper on the biocompatibility of mineral trioxide aggregate. BMC Oral Health 2016; 16: 111. 2. Beyazit Y, Kurt M, Kekilli M, Goker H, Haznedaroglu İC. Evaluation of Hemostatic Effects of Ankaferd as an Alternative Medicine. Altern Med Rev 2010; 15(4): 329-336. 3. Huri E, Haznedaroglu IC, Akgul T, Astarci M, Ustun H, Germiyanoulu C. Biphasic effects of ankaferd blood stopper on renal tubular apoptosis in the rat partial nephrectomy model representing distinct levels of hemorrhage. Saudi Med J. 2010; 31(8): 864-8. 4. Yılmaz E, Güleç Ş, Torun D, Haznedaroğlu İC, Akar N. The effects of Ankaferd Blood Stopper on transcription factors in HUVEC and the erythrocyte protein profile. Turk J Hematol 2011; 28: 276-85. 5. Arslan S, Haznedaroglu IC, Oz B, Goker H. Endobronchial application of Ankaferd blood stopper to control profuse lung bleeding leading to hypoxemia and hemodynamic instability. Respiratory Medicine 2009; 2: 144-6. 6. Kurt M, Akdogan M, Onal IK, Kekilli M, Arhan M, Aksu S, Kurt OK, Haznedaroglu IC. Endoscopic topical application of Ankaferd Blood Stopper for neoplastic gastrointestinal bleeding: a retrospective analysis. Dig Liver Dis 2010; 42: 196-9. 7. Lang I, Pabst M.A, Hiden U, Blaschitz A., Dohr G, Hahn T, Desoye G. Heterogeneity of microvascular endothelial cells isolated from human term placenta and macrovascular umbilical vein endothelial cells. Eur J Cell Biol 2003; 82(4): 163–173. 8. Ribatti D, Nico B, Vacca A, Roncali L, Dammacco F. Endothelial cell heterogeneity and organ specificity. J Hemototherap Stem Cell Res 2002; 11: 81–90. 9. Aird, W.C. Phenotypich heterogeneity of the endothelium: I.Structure, function, and mechanisms. Circ Res 2007; 100: 158–173. 10. Schechner JS, Nath AK, Zheng L, et al. In vivo formation of complex microvessels lined by human endothelial cells in an immunodeficient mouse. Proc Natl Acad Sci USA 2000: 97: 9191–9196.
Abstract
References
- 1. Dinçol ME, Ozbas H, Yılmaz B, Ersev H, Gokyay S and Olgac V. Effect of the plant-based hemostatic agent Ankaferd Blood Stopper on the biocompatibility of mineral trioxide aggregate. BMC Oral Health 2016; 16: 111. 2. Beyazit Y, Kurt M, Kekilli M, Goker H, Haznedaroglu İC. Evaluation of Hemostatic Effects of Ankaferd as an Alternative Medicine. Altern Med Rev 2010; 15(4): 329-336. 3. Huri E, Haznedaroglu IC, Akgul T, Astarci M, Ustun H, Germiyanoulu C. Biphasic effects of ankaferd blood stopper on renal tubular apoptosis in the rat partial nephrectomy model representing distinct levels of hemorrhage. Saudi Med J. 2010; 31(8): 864-8. 4. Yılmaz E, Güleç Ş, Torun D, Haznedaroğlu İC, Akar N. The effects of Ankaferd Blood Stopper on transcription factors in HUVEC and the erythrocyte protein profile. Turk J Hematol 2011; 28: 276-85. 5. Arslan S, Haznedaroglu IC, Oz B, Goker H. Endobronchial application of Ankaferd blood stopper to control profuse lung bleeding leading to hypoxemia and hemodynamic instability. Respiratory Medicine 2009; 2: 144-6. 6. Kurt M, Akdogan M, Onal IK, Kekilli M, Arhan M, Aksu S, Kurt OK, Haznedaroglu IC. Endoscopic topical application of Ankaferd Blood Stopper for neoplastic gastrointestinal bleeding: a retrospective analysis. Dig Liver Dis 2010; 42: 196-9. 7. Lang I, Pabst M.A, Hiden U, Blaschitz A., Dohr G, Hahn T, Desoye G. Heterogeneity of microvascular endothelial cells isolated from human term placenta and macrovascular umbilical vein endothelial cells. Eur J Cell Biol 2003; 82(4): 163–173. 8. Ribatti D, Nico B, Vacca A, Roncali L, Dammacco F. Endothelial cell heterogeneity and organ specificity. J Hemototherap Stem Cell Res 2002; 11: 81–90. 9. Aird, W.C. Phenotypich heterogeneity of the endothelium: I.Structure, function, and mechanisms. Circ Res 2007; 100: 158–173. 10. Schechner JS, Nath AK, Zheng L, et al. In vivo formation of complex microvessels lined by human endothelial cells in an immunodeficient mouse. Proc Natl Acad Sci USA 2000: 97: 9191–9196.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Health Care Administration |
| Journal Section | Case Report |
| Authors | |
| Publication Date | December 27, 2019 |
| Published in Issue | Year 2019 Volume: 9 Issue: 4 |