بررسی اثر حفاظتی عصاره گیاه بادرشبی (Dracocephalum moldavica L) بر القاي آپوپتوز و پرولیفراسیون انگل ليشمانيا ماژور در شرایط آزمایشگاهی
مهناز کریمی
1
(
1- دانشجوی دکتری میکروبیولوژی، دانشکده علوم پایه، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
)
الهه تاج بخش
2
(
گروه زیست شناسی دانشگاه آزاد اسلامی واحد شهرکرد
)
حسن ممتاز
3
(
استاد میکروبیولوژی، گروه میکروبیولوژی، واحد شهرکرد، دانشگاه آزاد اسلامی، شهرکرد، ایران
)
کلید واژه: آپوپتوز, ليشمانيا ماژور, گیاه بادرشبی, پرولیفراسیون,
چکیده مقاله :
با توجه به شیوع لیشمانیوز در مناطق مختلف جهان و ایران و نظر به اینکه ترکیبات پنج‌ظرفیتی آنتی‌موان برای درمان بیماری دارای عوارض جانبی متعددی می باشد، استفاده از گیاهان دارویی مورد تاکید قرار گرفته است. این تحقیق به منظور تعیین تاثیر عصاره گیاه دارویی بادرشبی (Dracocephalum moldavica L) بر روی آماستیگوت‌های لیشمانیا ماژور در شرایط آزمایشگاهی به روش رنگ سنجی انجام گرفت. در این مطالعه تجربی لیشمانیا ماژور (Lishmania Major) سویه استاندارد به محیط کشت RPMI1640 حاوی 10% سرم جنین گاوی FBS و آنتی‌بیوتیک‌هاي پنی‌سیلین- استرپتومایسین کشت داده شد و تا زمان رشد کامل در دماي 37 درجه و فشار 5 درصد CO2 انکوباتور در مدت 4-3 روز در شرایط استریل نگهداري شد. سپس در مرحله ثابت رشد تاثیر غلظت‌های مختلف عصاره‌های گیاه بادرشبی در مقایسه با داروی کنترل بر روی آماستیگوت‌های لیشمانیا ماژور در شرایط آزمایشگاهی با استفاده از روش رنگ سنجی MTT، مورد بررسی قرار گرفت. درصد انگل های زنده در حضور غلظت¬هاي مختلف عصاره و ترکیبات موثره بادرشبی پس از گذشت زمان های مختلف از کشت انگل کاهش چشمگیری داشتند. میزان IC50 عصاره و ترکیبات موثره بادرشبی در زمان هاي مختلف در مهار لیشمانیا ماژور اثر چشمگیری نشان داد.
چکیده انگلیسی :
Introduction: Due to the prevalence of leishmaniasis in different regions of the world and Iran and considering that pentavalent compounds of antimony have many side effects for the treatment of the disease; the use of medicinal plants has been emphasized. This research was conducted in order to determine the effect of the extract of the medicinal plant Dracocephalum moldavica L on Leishmania major Amastigote in laboratory conditions by colorimetric method. Methodology: In this experimental study, Leishmania Major standard strain was cultured in RPMI1640 culture medium containing 10% fetal bovine serum FBS and penicillin-streptomycin antibiotics and until full growth at 37 degrees temperature and 5% CO2 pressure in the incubator. It was kept in sterile conditions for 3-4 days. Then, in the stationary phase of growth, the effect of different concentrations of lemon balm plant extracts compared to the control drug on Leishmania major amastigote in laboratory conditions was investigated using the MTT colorimetric method. Results and conclusion: The percentage of live parasites in the presence of different concentrations of lemon balm extract and effective compounds decreased significantly after different times of parasite culture. The IC50 value of lemon balm extract and effective compounds at different times showed a significant effect in inhibiting Leishmania major.
1. Mougneau, E., F. Bihl, and N. Glaichenhaus, Cell biology and immunology of Leishmania. Immunological reviews, 2011. 240(1): p. 286-296.
2. Adler, S., Leishmania. Advances in parasitology, 1964. 2: p. 35-96.
3. Desjeux, P., Leishmaniasis: current situation and new perspectives. Comparative immunology, microbiology and infectious diseases, 2004. 27(5): p. 305-318.
4. Ivens, A.C., et al., The genome of the kinetoplastid parasite, Leishmania major. Science, 2005. 309(5733): p. 436-442.
5. Alrajhi, A.A., et al., Fluconazole for the treatment of cutaneous leishmaniasis caused by Leishmania major. New England Journal of Medicine, 2002. 346(12): p. 891-895.
6. Singh, N., et al., Natural product based leads to fight against leishmaniasis. Bioorganic & medicinal chemistry, 2014. 22(1): p. 18-45.
7. Kobets, T., I. Grekov, and M. Lipoldova, Leishmaniasis: prevention, parasite detection and treatment. Current Medicinal Chemistry, 2012. 19(10): p. 1443-1474.
8. Tajbakhsh, E., et al., The effects of medicinal herbs and marine natural products on wound healing of cutaneous leishmaniasis: A systematic review. Microbial Pathogenesis, 2021. 161: p. 105235.
9. Minodier, P. and P. Parola, Cutaneous leishmaniasis treatment. Travel medicine and infectious disease, 2007. 5(3): p. 150-158.
10. Zhan, M., et al., Dracocephalum moldavica L.: An updated comprehensive review of its botany, traditional uses, phytochemistry, pharmacology, and application aspects. Fitoterapia, 2023: p. 105732.
11. Dastmalchi, K., et al., Chemical composition and antioxidative activity of Moldavian balm (Dracocephalum moldavica L.) extracts. LWT-Food Science and Technology, 2007. 40(9): p. 1655-1663.
12. GOLPARVAR, A.R., et al., Chemical constituents of essential oil of Dracocephalum moldavica L. and Dracocephalum kotschyi Boiss. from Iran. Acta Agriculturae Slovenica, 2016. 107(1): p. 25-31.
13. Acimovic, M., et al., Chemical composition, antioxidant, and antimicrobial activity of Dracocephalum moldavica L. Essential oil and hydrolate. Plants, 2022. 11: p. 941.
14. Aslanipour, B., R. Heidari, and N. Farnad, Phenolic Combination and Comparison of Antioxidant Activity in Three Different Alcoholic Extracts of Dracocephalum moldavica L. Turkish Journal of Agriculture-Food Science and Technology, 2017. 5(3): p. 199-206.
15. مهسا, ک.ف., et al., بررسی اثر سمیت سلولی عصاره برگ تیمار شده بادرشبی (. Dracocephalum moldavica L) با استفاده از القاءکننده های مختلف بر روی چهار رده سلول سرطانی. 2020.
16. Murray, H., Interaction of Leishmania with a macrophage cell line. Correlation between intracellular killing and the generation of oxygen intermediates. The Journal of experimental medicine, 1981. 153(6): p. 1690-1695.
17. Geroldinger, G., et al., Techniques to study phagocytosis and uptake of Leishmania tarentolae by J774 macrophages. Experimental parasitology, 2019. 197: p. 57-64.
18. Rasti, S., et al., Comparison of molecular, microscopic, and culture methods for diagnosis of cutaneous leishmaniasis. Journal of clinical laboratory analysis, 2016. 30(5): p. 610-615.
19. Ilaghi, M., et al., The potential role and apoptotic profile of three medicinal plant extracts on Leishmania tropica by MTT assay, macrophage model and flow cytometry analysis. Parasite Epidemiology and Control, 2021. 12: p. e00201.
20. Junsi, M., et al., Efficacy of Thunbergia laurifolia (Rang Jued) aqueous leaf extract for specific biological activities using RAW 264.7 macrophage cells as test model. International Food Research Journal, 2017. 24(6): p. 2317-2329.
21. Garcia, A.R., et al., Cytotoxicity and anti-Leishmania amazonensis activity of Citrus sinensis leaf extracts. Pharmaceutical biology, 2017. 55(1): p. 1780-1786.
22. Dutta, A., et al., Development of a modified MTT assay for screening antimonial resistant field isolates of Indian visceral leishmaniasis. Parasitology international, 2005. 54(2): p. 119-122.
23. Najm, M., et al., Anti-Leishmanial Activity of Artemisia persica, A. spicigera, and A. fragrance against Leishmania major. Iranian journal of parasitology, 2021. 16(3): p. 464.
24. Alves, M.M.d.M., et al., Gallic and ellagic acids: two natural immunomodulator compounds solve infection of macrophages by Leishmania major. Naunyn-Schmiedeberg's archives of pharmacology, 2017. 390: p. 893-903.
25. Mendonça, D.V.C., et al., Antileishmanial activity of a naphthoquinone derivate against promastigote and amastigote stages of Leishmania infantum and Leishmania amazonensis and its mechanism of action against L. amazonensis species. Parasitology research, 2018. 117: p. 391-403.
26. Bezerra, E.A., et al., Garcinielliptone FC: Selective anti-amastigote and immunomodulatory effects on macrophages infected by Leishmania amazonensis. Toxicology in vitro, 2020. 63: p. 104750.
27. Sifaoui, I., et al., Activity of olive leaf extracts against the promastigote stage of Leishmania species and their correlation with the antioxidant activity. Experimental parasitology, 2014. 141: p. 106-111.
28. Sacks, D. and N. Noben-Trauth, The immunology of susceptibility and resistance to Leishmania major in mice. Nature reviews immunology, 2002. 2(11): p. 845-858.
29. Ahmed, S.B.H., et al., Evaluation of antileishmanial, cytotoxic and antioxidant activities of essential oils extracted from plants issued from the leishmaniasis-endemic region of Sned (Tunisia). Natural product research, 2011. 25(12): p. 1195-1201.
30. Ghaderi, A., et al., Evaluation of antileishmanial effect of the plant extract of alpha-pinene (Pistacia atlantica) in vitro and in vivo. Scientific Journal of Kurdistan University of Medical Sciences, 2018. 23(5): p. 32-44.
31. Tamargo, B., et al., In vitro and in vivo evaluation of essential oil from Artemisia absinthium L. formulated in nanocochleates against cutaneous leishmaniasis. Medicines, 2017. 4(2): p. 38.