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Burdock (Arctium Lappa) Leaf Extracts Increase the in Vitro Antimicrobial Efficacy of Common Antibiotics on Gram-Positive and Gram-Negative Bacteria

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Burdock (Arctium Lappa) Leaf Extracts Increase the in Vitro Antimicrobial Efficacy of Common Antibiotics on Gram-Positive and Gram-Negative Bacteria Open Chem., 2017; 15: 92–102 Research Article Open Access Lucia Pirvu*, Isabela Nicorescu, Cristina Hlevca, Bujor Albu, Valentin Nicorescu Burdock (Arctium lappa) Leaf Extracts Increase the In Vitro Antimicrobial Efficacy of Common Antibiotics on Gram-positive and Gram-negative Bacteria DOI 10.1515/chem-2017-0012 received January 23, 2017; accepted March 14, 2017. inhibitory) of Arctii folium extracts in combination with typical antibiotics as well as a potential use of the whole Abstract: This work aimed to study the potential effects of ethanol extract/W for restoring the antimicrobial potency four Arctii folium extracts, 5 mg gallic [GAE] acid equivalents of susceptible antibiotics have also been evidenced. per 1 mL sample, on six antibiotics (Ampicillin/AM, Tetracycline/TE, Ciprofloxacin/CIP, Sulfamethoxazole- Keywords: burdock leaves, interaction with usual Trimethoprim/SXT, Chloramphenicol/C and Gentamicin/ antibiotics, stimulatory and inhibitory effects CN) tested on four Gram-positive (Staphylococcus aureus ATCC 6538, Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 29212, and Staphylococcus 1 Introduction epidermidis ATCC 12228) and five Gram-negative (Proteus mirabilis ATCC 29245, Escherichia coli ATCC 35218, E. coli Arctium lappa L. (Asteraceae family), commonly greater ATCC 11229, E. coli ATCC 8739, and Bacillus cereus ATCC burdock, is a biennial species found across most of tEurope, 11778) bacteria. Arctii folium extracts were the whole Asia and also America. The root part, Bardanae radix, is ethanol extract/W and subsequent ethyl acetate/EA, used as a herbal medicine, but is also used as a vegetable aqueous/AQ, and chloroform/CHL fractions. Chemical in Eastern Asian regions (similar to potatoes) due to its qualitative analysis (HPTLC method) emphasized five nutritional value [1], the high content of carbohydrates main polyphenol compounds in Arctii folium polar (69%) including inulin (27-50%) and mucilage added to extracts: chlorogenic acid (Rf≈0.52/0.55) and its isomer, fats, proteins and numerous vitamins (B1, B2, B3, B5, B6, 1,5-di-O-caffeoylquinic acid (Rf≈0.90/0.92), plus cynarin B9, C, E and K) and minerals (Ca, Fe, Mg, Mn, P, K, Na and (Rf≈0.77), hyperoside (Rf≈0.68/0.64) and isoquercitrin Zn) [2] respectively. (Rf≈0.69/0.71). Microbiological screening indicated Arctii Concerning the basis of its medicinal use, data folium polar extracts (AQ and W) efficacy on S. epidermidis indicate that Bardanae radix is rich in caffeoylquinic acid ATCC 12228; the MIC values were in the range of common isomers [1, 3], with dicaffeoylquinic acid derivatives being antibiotics, being 32 and 128 μg GAE per mL sample quantified at 75.4% and 1,5-di-O-caffeoyl-4-O-maloylquinic respectively. The unpredictable effects (stimulatory or acid representing 44% of the total isomers [4], but that the content also comprisesnumerous other secondary metabolites such as tannin-iron complexes, various polyacetylenes and sulphuric actylene compounds, *Corresponding author: Lucia Pirvu: National Institute of Chemical- Pharmaceutical Research and Development (ICCF), Vitan 112, essential oils, guainolides and bitter compounds, lignans 031299,Bucharest, Romania, E-mail: [email protected] (e.g., arctigenin, arctiin,) and sterols (e.g., sitosterol, Isabela Nicorescu: Institute of Hygiene and Veterinary Public Health, stigmasterol) [2]. Studies regarding the metabolic profile Campul Mosilor 5, 021201, Bucharest, Romania of the compounds of burdock plant pieces indicated Cristina Hlevca, Bujor Albu: National Institute of Chemical- caffeoylquinic acid derivates and luteolin and quercetin Pharmaceutical Research and Development (ICCF), Vitan 112, rhamnosides occur in the root part, caffeoylquinic acid and 031299,Bucharest, Romania Valentin Nicorescu: University of Agronomic Sciences and Veterinary quercetin, quercitrin and luteolin derivates in the leaves, Medicine, Faculty of Veterinary Medicine, Splaiul Independentei whereas cynarin and chlorogenic acid are dominant in the 105, 050097, Bucharest, Romania seeds [5]. © 2017 Lucia Pirvu et al., published by De Gruyter Open. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License. Burdock Leaf Extracts Increase the In Vitro Antimicrobial Efficacy of Antibiotics on Bacteria 93 Based on this valuable phytochemical content, growing in Romania, this study was aimed to evaluate any Bardanae radix represents an important ingredient potential interaction of Arctii folium extracts with typical for traditional medicine practitioners, and has also antibiotics. The results of the study are very useful to avoid achieved international recognition for its numerous situations of incompatibility with such antibiotics, but also healthy effects [1]; Romanian folk medicine also considers as the starting point for developing new natural synergistic a bardanae radix hot water extract (decoction type) products able to restore antimicrobic susceptibility. as an effective expectorant, anti-tussive, emollient, diuretic and anti-inflammatory remedy thus being recommended for different lung, digestive, renal and skin ailments [6]. Proving these, studies have reported 2 Experimental Procedure burdock products and corresponding separate extracts or specific compounds (especially lignans such as arctiin 2.1 Materials and arctigenin, and polyphenols compounds) having antioxidant, anti-inflammatory, antitumor, antibacterial 2.1.1 Plant material description and antiviral activities [1], as well as hepatoprotective [7], antiurolithiasic [8] and skin improving [1, 9] effects, with Leaves from burdock (Arctii folium) were harvested in some clinical studies also proving their effectiveness in June and September from the Romanian Carpathians, knee osteoarthritis [10] and for dental infectious conditions Sinaia region (1,000 m altitude). Taxonomic identification [11, 12]; burdock root and fruits are also reported to have was carried out by the botanist’s team of the National antidiabetic activity [1, 4], with their beneficial effects Institute of Chemical-Pharmaceutical R&D (ICCF being attributed to the copresence of sitosterol, inulin Bucharest), Romania. The burdock leaves were shade and lignans[2]. Furthermore, the combination arctigenin dried and ground to a medium-size plant powder; voucher - polyphenols have shown the capacity to block tumor cell specimens (ALam7.1, Alam7.2) are deposited in the ICCF growth and metastasis [13]. Additionally, arctiin, caffeic Plant Material Storing Room. and chlorogenic acid mixtures have proven significant anti-mutagenic effects and a positive correlation with polyphenolic content [14]. Concerning the biochemical 2.1.2 Vegetal extracts preparation pathway involved, studies on arctigenin tested on MH60 cells indicated the stimulation of apoptosis (IC50 = 1.0 μM) Fifty (50) g of leaf powder (Arctii folium) were heat-assisted as being the most probable mechanism [15], while the extracted (twice) with 500 mL of 70% (v/v) ethanol. The capacity of polyphenols to protect the cells against radical total extract obtained was filtered through a medium( ) oxygen species (ROS) injuries and to stimulate immune paper filter, resulting in 650 mL ethanol extract (E7). Two processes [16] are believed to effectively contribute to the hundred and fifty (250) mL of (E7) ethanol extract was antitumor effect of burdock products. concentrated at low pressure at sicc product then passed The leaves of Arctium lappa L. have been recognized into 20% (v/v) propylene glycol (20% PG) solution so as havingsome antimicrobial properties against some as to ensure a final concentration of exactly 5 mg total bacterial strains found in endodontic infections such phenols (expressed as gallic acid [GAE] equivalents) per as Bacillus subtilis, Lactobacillus acidophilus and 1 mL extract, further called Arctii folium whole ethanol Pseudomonas aeruginosa, as well as on Candida albicans, extract (codified W); the whole ethanol extract (W) was Escherichia coli, Staphylococcus aureus and Micrococcus divided into 2 mL Eppendorf tubes and stored at -8 °C until luteus [12]. Furthermore, the efficacy of burdock leaf microbiological studies. Another two hundred and fifty derived products against influenza A virus (A/NWS/33, (250) mL of (E7) ethanol extract was also concentrated at H1N1) (IFV) [17], herpesvirus (HSV-1, HSV-2), adenovirus sicc product then dissolved into 100 mL of distilled water. (ADV-3, ADV-11), and human immunodeficiency virus The resulting aqueous solution was (manually) extracted (HIV-1) [18] has also been proven. first with (3 x 100 mL) chloroform and then with (3 x 100 Thus, in the current upward trend of using plant mL) ethyl acetate solvent, 24 hours per each stage. Three derived products as natural medicines, while at the same selective extracts were obtained: aqueous (AQ), ethyl time increasing bacterial resistance to antibiotics of all acetate (EA) and chloroform (CHL) fractions, respectively. classes, based on these data, but also on our previous The three Arctii folium selective fractions were analyzed as results [19, 20] demonstrating the antimicrobial effects of concerning total phenols content (GAE per 1 mL sample) some whole and selective extracts from medicinal plants then concentrated at sicc product and (separately) passed 94 Lucia Pirvu et al. into 20% PG as follows: aqueous and ethyl acetate ATCC 29245, Escherichia coli ATCC 35218, E. coli ATCC fractions were prepared so as to achieve identical 5 mg 11229, E. coli ATCC 8739, and Bacillus cereus ATCC 11778).
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