Comparison of antioxidant, anti-inflammatory, antimicrobial activity and chemical composition of aqueous and hydroethanolic extracts of the herb of Tropaeolum majus L.
Graphical abstract
Introduction
Tropaeolum majus L. (T. elatum Salisb., Tropaeolaceae) comes from the mountainous areas of South and Central America. Nasturtium is native to Peru and Bolivia. It has been grown in Europe since the seventeenth century due to its uses in herbal medicine and as an ornamental plant. Phytochemical studies of T. majus have reported the occurrence of sulfur compounds (glucotropaeolin), flavonoids (quercetin and kaempferol glycosides), anthocyanins (delphinidin, cyanidin and pelargonidin derivatives), carotenoids (lutein, zeaxanthin, β-carotene), phenolic acids (chlorogenic acid), cucurbitacines and vitamin C (Bruneton, 1999, Garzón and Wrolstad, 2009, Niizu and Rodriguez-Amaya, 2005, Strzelecka and Kowalski, 2000, Wojciechowska and Wizner, 1983).
Nasturtium herb is mentioned in pharmacognosy and phytotherapy textbooks as treatment for upper respiratory tract (tonsillitis, bronchitis) and urinary tract diseases (Bruneton, 1999, Fournie, 1947, Strzelecka and Kowalski, 2000). Moreover, it is used externally in dermatology and cosmetology for the treatment of diseases of the skin, nails and hair (dandruff with itching and peeling), sunburns, superficial and limited burns and diaper rashes (Bruneton, 1999). Nasturtium herb and extracts have been used for years in folk medicine in many countries, an extract of nasturtium is ingredient of a medicinal product Angocin® Anti-Infekt N and dermocosmetics (anti-dandruff shampoo). Furthermore, nasturtium due to the high content of biologically active compounds is a valuable component in the diet and can be a raw material for the production of nutraceuticals. Nasturtium is also used in the kitchen. Dishes containing nasturtium have a sharp, spicy taste. In many countries fresh leaves are eaten in salads. Marinated in vinegar undeveloped flower buds are used in dishes as a substitute for capers, whereas chopped unripe fruits can be added to tartar sauce instead of horseradish. Nasturtium flowers are added to soups, meats, pasta dishes and paste spreads. They can also be fried in pancake batter. In Canada, nasturtium blossoms stuffed with cottage cheese are very popular. Moreover, leaves, stems and flowers are used for the preparation of herbal vinegar (Grzeszczuk et al., 2010, Mikołajczyk and Wierzbicki, 1989).
One of the main activities of the herb is its antimicrobial activity, which is explained by the presence of benzyl isothiocyanate (a breakdown product of glucotropaeolin) (Virtanen, 1969). Antimicrobial activity of benzyl isothiocyanate has been widely documented (Jang et al., 2010, Kamii and Isshiki, 2009). However by contrast there is little research on the antimicrobial activity of extracts obtained from the nasturtium herb. Activity against Staphylococcus epidermidis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Salmonella setubal by hexane and chloroform fractions obtained from 70% ethanolic extract of aerial parts of nasturtium has been shown by Zanetti et al. (2003). Natriuretic and diuretic activity of hydroethanolic extract of nasturtium leaves (90% ethanol as a solvent to maceration was used), ethanolic fraction obtained from the hydroethanolic extract and isoquercitrin, have been shown by Gasparotto et al., 2009, Gasparotto et al., 2011a. Those results support the use of T. majus as a diuretic in diseases of the urinary tract. Gasparotto et al. (2011b) have also shown an antihypertensive effect of the same extracts and isoquercitrin. Furthermore, the same team have demonstrated the absence of subchronic toxicity, no (anti)estrogenic or (anti)androgenic activity, and no effect on uterine contractility in rats after treatment with hydroethanolic extract of the nasturtium herb (Gomes et al., 2012, Lourenço et al., 2012). For benzyl isothiocyanate and raw materials derived from nasturtium anticoagulant activity has also been shown (De Medeiros et al., 2000, Santo et al., 2007). Moreover antioxidant activity against synthetic radicals (ABTS, DPPH) has been demonstrated for the nasturtium flowers (Garzón and Wrolstad, 2009).
It is known that ROS and RNS are involved in the pathophysiology of inflammation. They entering the human body extraneous matter stimulate neutrophils and macrophages, which migrate to the area of inflammation (Halliwell, 1987, Pacher et al., 2007). Because of the great importance of ROS and RNS in the pathogenesis of inflammation, and because the nasturtium herb is recommended in many inflammatory diseases (also caused by pathogens) it is advisable to examine the antioxidant, anti-inflammatory and antimicrobial activities of this plant.
The aim of our study was comparison of antioxidant, anti-inflammatory, antimicrobial activity and chemical composition of extracts of T. majus L. herb. In the first part of the work, comparison of the activity of aqueous and hydroethanolic extracts derived from dried and fresh (freeze-dried) nasturtium herb prepared both at room temperature and at 90 °C, was made. The second part of the work included an analysis of the composition of the extracts, plus a determination of the content of benzyl isothiocyanate and ascorbic acid. The antioxidant activity was evaluated by examining the scavenging of synthetic radical (DPPH) and five radicals (O2−, H2O2, HClO, NO and ONOO−) generated in cell-free systems, as well as in ex vivo experiment (oxidant generation by f-MLP-stimulated neutrophils). To examine the anti-inflammatory activity of the studied extracts, inhibition of hyaluronidase and cyclooxygenase 1 (COX1) activity by the extracts was tested. Studies of composition of the extracts were performed using high-performance thin layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC) coupled with diode array detector (DAD) and ion trap mass detector. The content of benzyl isothiocyanate in the tested extracts was determined by gas chromatography (GC), and the content of ascorbic acid was studied using the colorimetric method.
Section snippets
Chemicals
Bovine testis hyaluronidase (BTH), 4,5-diaminofluorescein (DAF-2), 2-diphenyl-1-picrylhydrazyl radical (DPPH), 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB), formyl-met-leu-phenylalanine (f-MLP), Hanks’ balanced salt solution (HBSS), horseradish peroxidase (HRP), hydrogen peroxide (H2O2), indomethacin, luminol, nitrobluetetrazolium (NBT), 3-morpholinosydnonimine hydrochloride (SIN), xanthine, and xanthine oxidase were purchased from Sigma–Aldrich Chemie GmbH (Steinheim, Germany). NaBH4 was
Results and discussion
Nasturtium herb is used as treatment for upper respiratory tract and urinary tract diseases, externally in dermatology and cosmetology for the treatment of diseases of the skin, nails and hair (dandruff with itching and peeling), sunburns, superficial and limited burns and diaper rashes, and is also used in the kitchen. Moreover the herb is a valuable component in the diet and can be a raw material for the production of nutraceuticals. In medicine and cosmetology, the most popular extracts are
Acknowledgments
This project was carried out with the use of CePT infrastructure financed by the European Union under the European Regional Development Fund within the Operational Programme “Innovative economy”; for 2007–2013. The authors would like to thank Mrs. Monika Marciniak, Ms. Natalia Szyller and Mr. Roman Rudecki for their technical assistance.
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