Investıgatıon of the Antimicrobial and Apoptotic Efficacy of Hyoscyamus retıculatus L. Seed Extract

Öz

Hyoscyamus reticulatus is from Solanacea famil and has six species in our country. Hyoscyamus species are the source of tropan alkaloids, such as scopolamine and hyoctiamine, which are commonly used in public. The aim of this study was to determine the antimicrobial activity of ethyl alcohol extract obtained from H. reticulatus seed by soxhlet method against some Gram (+) and Gram (-) bacteria and yeasts and to determine apoptotic effect of H. reticulatus seed by measuring TP53 and Bcl-2 levels in plasma and brain samples. 40 Wistar albino female rats were divided into 4 groups in each group with 10 of them; Control (C), 125, 250, 500 mg / kg H. reticulatus extract groups. It was found that H. reticulatus seed extract have no antimicrobial activity against Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pasteurella multicoda, Pseudomonas aeruginosa, Salmonella enteridis, Staphylococcus aureus, Yersinia enterocolitica bacteria and Candida albicans yeast. Plasma samples of HR250 and HR500 groups showed significant increase in TP53 levels compared to group C (p <0.01, p <0.001, respectively). Compared to all extract applied groups and group C; Plasma Bcl-2 levels significantly decreased in extract groups (p <0.001). As a result; we suggest that H. reticulatus seed extract does not have an antimicrobial effect and extracts applied in different doses in vivo may stimulate apoptosis in other organ and / or organs, although not in brain tissue.

Anahtar Kelimeler

• Antimicrobial
• Bcl-2
• Hyoscyamus reticulatus seed extract
• TP53

Hyoscyamus reticulatus L. Tohum Ekstraktının Antimikrobiyal ve Apoptotik Etkinliğinin Araştırılması

Öz

Hyoscyamus reticulatus Solanacea ailesindendir ve ülkemizde altı türü bulunmaktadır. Hyoscyamus türleri özellikle halk arasında yaygın olarak kullanılan skopolamin ve hiyosiyamin gibi tropan alkaloitleri kaynağıdır. H. reticulatus tohumundan soxhlet yöntemi ile elde edilen etil alkol ekstresinin bazı Gram (-) ve Gram (+) bakteri ve mayalara karşı antimikrobiyal aktivitesinin tespit edilmesi ve H. reticulatus tohumunun plazma ve beyin numunelerinde TP53 ve Bcl-2 seviyeleri ölçülerek apoptotik etkisinin belirlenmesi amaçlanmıştır. Çalışmada 40 adet Wistar albino cinsi dişi sıçan her grupta 10 adet olacak şekilde 4 gruba ayrıldı; Kontrol (K), 125, 250, 500 mg/kg H. reticulatus ekstrakt grupları. H. reticulatus tohumu ekstraktının Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Pasteurella multicoda, Pseudomonas aeruginosa, Salmonella enteridis, Staphylococcus aureus, Yersinia enterocolitica bakterileri ve Candida albicans mayasına karşı antimikrobiyal etkinliğinin bulunmadığı tespit edilmiştir. HR250 ve HR500 gruplarının plazma örneklerinde TP53 seviyelerinin K grubuna göre önemli düzeyde arttığı tespit edildi (sırasıyla p<0,01, p<0,001). Ekstrakt uygulanan tüm gruplarla K grubu karşılaştırıldığında; ekstrakt uygulanan gruplarda plazma Bcl-2 seviyelerinin önemli düzeyde azaldığı belirlendi (p<0,001). Sonuç olarak; H. reticulatus tohum ektraktının antimikrobiyal etkisi olmadığı ve in vivo olarak farklı dozlarda uygulanan ekstraktın beyin dokusunda olmasa da diğer organ ve/veya organlarda apoptozisi uyarabileceğini düşünmekteyiz.

Anahtar Kelimeler

• Antimikrobiyal
• Bcl-2
• Hyoscyamus reticulatus tohum ekstraktı
• TP53

Kaynakça

1. [1] Baytop, T. 1999. Türkiye’de Bitkilerle Tedavi. 2.baskı. Nobel Tıp Kitabevleri, İstanbul, 480s.
2. [2] Madani, H., Hosseini, B., Dehghan, E., Rezaei-chiyaneh, E. 2015. Enhanced production of scopolamine in induced autotetraploid plants of Hyoscyamus reticulatus L. Acta Physiologiae Plantarum, 37(3), 55.
3. [3] Ghorbanpour, M., Hatami, M., Hatami, M. 2015. Activating antioxidant enzymes, hyoscyamine and scopolamine biosynthesis of Hyoscyamus niger L. plants with nano-sized titanium dioxide and bulk application. Acta Agriculturae Slovenica, 105, 23–32.
4. [4] Evans, W. C., Ramsey, K. P. A. 1983. Alkaloids of the Solanaceae tribe Anthocercideae. Phytochemistry, 22(10), 2219-2225.
5. [5] Bahmanzadegan, A., Sefidkon, F., Sonboli, A. 2010. Determination of hyoscyamine and scopolamine in four Hyoscyamus species from Iran. Iranian Journal of Pharmaceutical Research, 65-70.
6. [6] Karadaş, S., Güler, A., Şahin, M., Behçet, L. 2012. 32 haftalık gebede banotu zehirlenmesi. Van Tıp Dergisi, 19(1), 36-38.
7. [7] Kürkçüoğlu, M. 1970. Henbane (Hyoscyamus niger) poisoning in the vicinity of Erzurum. The Turkish Journal of Pediatrics, 12(1), 48-56.
8. [8] Güler, G. Ö. 2012. Studies on antioxidant properties of the different solvent extracts and fatty acid composition of Hyoscyamus reticulatus L. Journal Food Biochemistry, 36(5), 532-538.

9. [9] Singh, S. K., Pandey, V. D., Evaluation of Hyoscyamus niger L. extracts for antibacterial activity. Plant Archives, 9(1), 97-100.
10. [10] Dulger, B., Goncu, B. S., Gucin, F. 2010. Antibacterial activity of the seeds of Hyoscyamus niger L. (Henbane). Asian Journal of Chemistry, 22(9), 6879-6883.
11. [11] Erkal, H., Özyurt, Y., Arıkan, Z. 2006. Yaşlı hastada henbane (Hyoscyamus niger) sonrası antikolinerjik sendrom. Turkish Journal of Geriatrics, 9(3), 188-191.
12. [12] Straus, A. 1989. Hyoscyamus spp: In vitro culture and production of tropane alkaloids. ss 286–314. In Medicinal and Aromatic Plants II, Springer, Berlin Heidelberg New York, 538s.
13. [13] Yılmaz, O. 1990. Bursa yöresinde yetişen önemli zehirli bitkilerin toksikolojik özellikleri. Uludağ Üniversitesi, Sağlık Bilimleri Enstitüsü, Doktora Tezi, 93s, Bursa.
14. [14] Kartal, M., Kurucu, S., Altun, L., Ceyhan, T., Sayar, E., Cevheroğlu, S., Yetkin, Y. 2003. Quantitative analysis of l-hyoscyamine in Hyoscyamus reticulatus L. By GC-MS. Turkish Journal of Chemistry, 27(5), 565-569.
15. [15] Gross, A., McDonnell, J. M., Korsmeyer, S. J. 1999. BCL-2 family members and the mitochondria in apoptosis. Genes & Development, 13(15), 1899-1911.
16. [16] Certo, M., Moore, V. D. G., Nishino, M., Wei, G., Korsmeyer S., Armstrong S. A., Letai A. 2006. Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell, 9(5), 351-365.
17. [17] Martinou, J. C., Youle, R. J. 2011. Mitochondria in apoptosis: Bcl-2 family members and mitochondrial dynamics. Developmental Cell, 21(1), 92-101.
18. [18] Coşkun, G., Özgür, H. 2014. Apoptoz ve nekrozun moleküler mekanizması. Arşiv Kaynak Tarama Dergisi, 20(3), 145-158.
19. [19] Fridman, J. S., Lowe, S. W. 2003. Control of apoptosis by p53. Oncogene, 22(56), 9030-9040.
20. [20] Murray, P. R., Baron, E., Pfaller, M., Tenover, F. C., Yolke, R. H. 1995. Manual Clinical Microbiology. ASM, Washington DC, 1356s.
21. [21] Sacchetti, G., Maietti, S., Muzzoli, M,, Scaglianti, M., Manfredini, S., Radice, M., Bruni, R. 2005. Comparative evaluation of 11 essential oils of different origin as functional antioxidants, antiradicals and antimicrobials in foods. Food Chemistry, 91(4), 621-632.
22. [22] Kumar, A., Gupta, R. S. 1984. A note on the sensitivity of enterotoxigenic Staphylococcus aureus for garlic extract (Allium sativum Linn). Indian Veterinary Journal, 61(1), 718-719.
23. [23] Chalabian, F., Majd, A., Mehrabian, S., Falahian, F. 2002. A study of growth inhibitory effect of alkaloids of two species of genus hyoscyamus on some kinds of microbes of skin. Journal of Scıences, 12(43), 3371-3378.
24. [24] Bazzaz, B., Haririzadeh, G. 2003. Screening of Iranian plants for antimicrobial activity. Pharmaceutical Biology, 41(8), 573-583.
25. [25] Yücel Mercan, U., Yılmaz, O. 2014. Van Yöresinde Yetişen İki Hyoscyamus (Banotu) Türünün Total Alkaloid Miktarları. Van Veterinary Journal, 25(3), 77-80.
26. [26] Chatterjee, S. K., Nandi, R. P., Baharati, P., Yonjan, M. K. 1988. Improvement studies on some alkaloid yielding medicinal plants. Medicinal and Aromatic Spice Plants, 180(1), 39-46.
27. [27] Poutaraud, A., Girardin, P. 2005. Influence of chemical characteristics of soil on mineral and alkaloid seed contents of Colchicum autumnale L. Environmental and Experimental Botany, 54(2), 101-108.
28. [28] Nejadhabibvash, F., Rahmani, F., Heidari, R., Jamei, R., Azimi, F. 2012. Study of inheritance and environment on tropane alkaloids within Hyoscyamus species. Australian Journal of Crop Science, 6(10), 1428-1434.
29. [29] Haupt, S., Berger, M., Goldberg, Z., Haupt, Y. 2003. Apoptosis-the p53 network. Journal of Cell Science, 116(20), 4077-4085.