ارزیابی مقدار تاکسول و 10-دِ استیل باکاتین III در برگ و کشت سوسپانسیون سلولی دو گونه سرخدار

نوع مقاله: علمی - پژوهشی

نویسندگان

1 دانش‌آموخته کارشناسی ارشد فیزیولوژی و اصلاح گیاهان دارویی، گروه کشاورزی، پژوهشکده گیاهان و مواد اولیه دارویی دانشگاه شهید بهشتی، تهران، ایران

2 دانشیار، گروه کشاورزی، پژوهشکده گیاهان و مواد اولیه دارویی دانشگاه شهید بهشتی، تهران، ایران

3 استادیار، گروه کشاورزی، پژوهشکده گیاهان و مواد اولیه دارویی دانشگاه شهید بهشتی، تهران، ایران

4 استادیار، گروه فیتوشیمی، پژوهشکده گیاهان و مواد اولیه دارویی دانشگاه شهید بهشتی، تهران، ایران

چکیده

در این تحقیق، مقدار تاکسول و 10-دِ ‌استیل باکاتین III در برگ‌های گونه سرخدار بومی ایران (Taxus baccata) و گونه اقیانوس آرام (T. brevifolia) و توانایی تولید آن‌ها در کشت سوسپانسیون سلولی این دو گونه مورد بررسی قرار گرفت. این تحقیق در سال 1392 در پژوهشکده گیاهان دارویی دانشگاه شهید بهشتی تهران اجرا گردید. القای کالوس با استفاده از ریزنمونه‌های میان‌گره ساقه‌های جوان در محیط کشت جامد صورت گرفت، سپس کشت سوسپانسیون سلولی بر روی همین محیط کشت حاوی تنظیم‌کننده‌های رشد نفتالن استیک اسید و بنزیل‌آمینوپورین به‌ترتیب با مقادیر 2 و 1/0 میلی‌گرم در لیتر ایجاد شد. 21 روز پس از کشت، وزن تر و خشک سلول محاسبه و مقدار تاکسول و 10-دِ‌ استیل ‌باکاتین III آن‌ها توسط دستگاه کروماتوگرافی مایع با عملکرد بالا اندازه‌گیری شد. نتایج نشان داد که برگ‌های گونه های سرخدار بومی ایران و سرخدار اقیانوس آرام به‌ترتیب حاوی 45 و 27 میکروگرم تاکسول و 40 و 4 میکروگرم 10-دِ ‌استیل ‌باکاتین III در گرم وزن خشک نمونه بودند و کشت سوسپانسیون سلولی آن‌ها نیز به‌ترتیب مقدار540 و 20 میکروگرم در لیتر تاکسول و مقدار 1 و 110 میکروگرم در لیتر از 10-دِ ‌استیل ‌باکاتین III تولید کردند. بنابراین نتایج حاکی از برتری کشت سوسپانسیون سلولی سرخدار بومی ایران در تولید تاکسول بود که می‌تواند برای بهینه سازی تولید تجاری این ترکیب ضد سرطان در سیستم های بیوراکتور مورد استفاره قرار گیرد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Quantification of Taxol and 10-Deacetyl Baccatin III in the Leaf and Cell Suspension Cultures of Two Taxus L. Species

نویسندگان [English]

  • H. Ahadi 1
  • M.H. Mirjalili 2
  • M. Farzaneh 3
  • H. Rezadoost 4
1 M.Sc. Graduate of Medicinal and Aromatic Plants, Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Tehran, Iran
2 Associate Professor, Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Tehran, Iran
3 Assistant Professor, Department of Agriculture, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Tehran, Iran
4 Assistant Professor, Department of Phytochemistry, Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, G. C., Tehran, Iran
چکیده [English]

Abstract
Background and Objectives
Taxanes are a family of diterpene alkaloids which is currently known as the most important anticancer compounds to suppress and inhibit cell growth, differentiation and proliferation in indefinitely known cancer cell lines. The most well recognized member of the taxane family is taxol (paclitaxel), a minor cytotoxic component derived originally from yew bark (Taxus brevifolia). 10-Deacetylbaccatin III (10-DAB), which is a homolog of taxol, can be extracted at high yields from the needles of Taxus baccata and is currently considered the most appropriate precursor for semi-synthesizing taxol and its analogue, taxotere (docetaxel). The natural source of both compounds is the inner bark and needles of several Taxus species, especially T. baccata and T. brevifolia. Plant cell culture is an environmentally sustainable source of taxol and offers several advantages as it is not subjected to weather, season or contamination. The aim of the present study was to evaluate taxol and 10-DAB content of two Taxus species and their ability in production of both taxanes through cell culture.
Materials and Methods
The plant materials of T. baccata and T. berevifolia were collected from Shahid Beheshti University (35° 48' N, 51° 23' E at an altitude of 1785 m) and botanical garden of Faculty of Agriculture, University of Tehran, Karaj (35° 48' N, 50° 59' E at an altitude of 1238 m), respectively. Callus culture of both species was initiated from internodal segments on B5 medium supplemented with plant growth reaulators. Cell suspension culture was established on the medium supplemented with 2.0 mg L−1 naphtalene acetic acid (NAA) and 0.1 mg L−1 benzyl amino purine (BAP). Cell viability was determined by Evan’s blue staining test. After 21 days, the amount of taxol and 10-DAB was analysed by HPLC.
Results
The results showed that mother stocks of T. baccata and T. brevifolia contain 45 and 27 µg/g DW of taxol and 40 and 4 µg/g DW of 10-DAB, respectively. Cell growth was estimated by measuring fresh weight (FW) and dry weight (DW). Results showed that the maximum growth was reached after 21 days of cultivation. Cell cultures of T. baccata and T. brevifolia produced 540 and 20 µg/l taxol and 1 and 110 µg/l of 10-DAB, respectively. Taxol production in T. baccata cell culture was 26-fold higher than in T. berevifolia cell culture.
Discussion
As far as our literature survey could ascertain, growth and taxol production in T. baccata cell cultures have previously been reported. Although, T. baccata cells are more potent for the production of taxol, T. brevifolia cell culture can be considered for the production of 10-DAB, a homolog of taxol, as an appropriate precursor for semi-synthesizing taxol and its analogue, taxotere (docetaxel). However, it is essential to study and quantify the effect of selected key medium components on growth as well as product accumulation and strike a balance between the two to enhance the yield and productivity.

کلیدواژه‌ها [English]

  • Anticancer
  • In vitro culture
  • Liquid chromatography
  • Taxane
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