|Year : 2020 | Volume
| Issue : 72 | Page : 750-756
Chemical composition and biological potential of a chloroform fraction from the leaves of marine plant Syringodium filiforme Kützing
Kethia Gonzalez Garcia1, David Marrero Delange1, Yasnay Hernández Rivera1, Yulexi Acosta Suárez1, Richard Gutiérrez Cuesta1, Mario Riera-Romo2, Olga Echemendia3, Lívia Macedo Dutra4, Jackson Roberto Guedes Da Silva Almeida4, Dayana Pérez-martínez5, Laurent Picot6, Idania Rodeiro Guerra2
1 Department of Chemistry, Institute of Marine Science, Revolución, Havana, Cuba
2 Department of Pharmacology, Institute of Marine Science, Havana, Cuba
3 Carlos J. Finlay Research Institute, Havana, Cuba
4 Department of Pharmacy, Federal University of Vale Do São Francisco, Petrolina, Pernambuco, Brazil
5 Center For Molecular Immunology, Directorate of Tumor Immunology, Havana, Cuba
6 Faculty of Science and Technology, UMRI CNRS Lienss, La Rochelle University, La Rochelle, France
|Date of Submission||14-Feb-2020|
|Date of Decision||13-Apr-2020|
|Date of Acceptance||01-Dec-2020|
|Date of Web Publication||16-Feb-2021|
David Marrero Delange
Loma 14 and 37 St., Alturas Del Vedado, Plaza De La Revolucion, Havana 10600
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Seagrasses are an important component of Nearshore marine ecosystems and a rich source of secondary metabolites with important pharmacological properties. Materials and Methods: In this work, crude hydroethanolic extract and chloroform fraction (SfCHCl3) from marine plant Syringodium filiforme were evaluated for antimicrobial and cytotoxic potentials. In addition, the chemical composition of chloroform fraction was determined by gas chromatography-mass spectrometry (GC-MS) analysis. Results: GC-MS analysis allowed the identification of 68 compounds in the SfCHCl3, where palmitic acid (39.18%) was the main component. The evaluation of antibacterial activity of SfCHCl3 showed good activity against strains of Staphylococcus aureus (ATCC 6538), Pseudomonas aeruginosa (ATCC 9027), and Salmonella typhi (ATCC 9992v), and it was less active against Escherichia coli (ATCC 10576) and Candida albicans (ATCC 10231), while the crude extract showed low antibacterial activity. Furthermore, an important cytotoxic effect of SfCHCl3 in the A549 human lung carcinoma cell line was evidenced, which were similar to the cytotoxicity of cisplatin in the same cell line. Conclusion: The results suggest that compounds may play an important role in the antimicrobial and cytotoxic effects observed for this species. In addition, this research contributes to the chemotaxonomic characterization of S. filiforme and validates this species as a potential source of natural antimicrobial and cytotoxic molecules.
Keywords: Syringodium filiforme, Chemical composition, Chloroform fraction, Antimicrobial, Cytotoxicity
|How to cite this article:|
Garcia KG, Delange DM, Rivera YH, Suárez YA, Cuesta RG, Riera-Romo M, Echemendia O, Dutra LM, Da Silva Almeida JR, Pérez-martínez D, Picot L, Guerra IR. Chemical composition and biological potential of a chloroform fraction from the leaves of marine plant Syringodium filiforme Kützing. Phcog Mag 2020;16:750-6
|How to cite this URL:|
Garcia KG, Delange DM, Rivera YH, Suárez YA, Cuesta RG, Riera-Romo M, Echemendia O, Dutra LM, Da Silva Almeida JR, Pérez-martínez D, Picot L, Guerra IR. Chemical composition and biological potential of a chloroform fraction from the leaves of marine plant Syringodium filiforme Kützing. Phcog Mag [serial online] 2020 [cited 2021 Feb 28];16:750-6. Available from: http://www.phcog.com/text.asp?2020/16/72/750/309308
- Crude hydroethanolic extract and chloroform fraction (SfCHCl3) from marine plant Syringodium filiforme were evaluated for antimicrobial and cytotoxic potentials.
- GC-MS analysis allowed the identification of 68 compounds in the SfCHCl3, being palmitic acid (39.18 the predominant substance.
- This study allowed demonstrating antimicrobial and cytotoxic effects of the species S. filiforme.
Abbreviations used: SfCHCl3: Chloroform fraction of Syringodium filiforme; GC-MS: Gas chromatography-mass spectrometry.
| Introduction|| |
Marine species are constantly competing for extent their habitats. To survive to this intense competition and such extreme conditions, seagrasses and algae have biosynthetically developed a wide variety of secondary metabolites. These wide range of unique chemical entities have lately drawn the attention of investors from worldwide pharmaceutical companies because of their broad panel of pharmacological activities and their health benefits.,,,,,,,
In this sense, the presence of diverse potent secondary metabolites in seagrasses is well documented,,,, while their antibacterial, anticancer, antiproliferative, antitumor, and antioxidant effects make them being recognized as new potential drugs.,,,
Previous investigations, only a few studies, have been conducted about the chemical composition and pharmacological activities of Syringodium genus. Nussier et al. identified in the hydroethanolic extract of Syringodium filiforme (manatee grass), phenolic compounds such as p-hydroxybenzoic, vanillic, caffeic, protocatechuic, p-coumaric, ferulic, syringic, chicoric, and caftaric acids. From Syringodium flotsam, L-chiro-inositol was isolated and characterized, with demonstrated hypoglycemic action. The qualitative analysis of extracts from this species showed high content of flavonoids, phenols, terpenes, anthocyanins, reduced sugars, and alkaloids. From all evaluated extracts, only the total extract and methanol fraction revealed asignificant antioxidant properties against free radicals.
Similarly, a qualitative test of phytochemicals from methanol extracts of S. isoetifolium showed the occurrence of phytoconstituents such as saponins, resins, proteins, carbohydrates, glycosides, acidic compounds, reduced sugars, cardiac glycosides, phenols, and alkaloids. The antibacterial activity of methanol and acetone extracts from this species against 17 human pathogens and five fish pathogens were demonstrated, while the lipophilic fraction from S. filiforme was active against Schizochytrium aggregatum and Pseudoalteromonas bacteriolytica.
On the other hand, the cytotoxic activity has been less explored for the Syringodium genus. There are reports of the cytotoxicity of hydroethanolic extracts from other marine plants such as Thalassodendron ciliatum (Egyptian Red Sea seagrass) and Thalassia testudinum (turtle grass)., In addition, the cytotoxicity of a chloroform fraction recently obtained from T. testudinum was assessed in A549 human lung carcinoma. S. isoetifolium, together with other marine angiosperms such as Cymodocea serrulata have demonstrated potent antifouling and antimicrobial activity against microalgae and pathogenic bacteria. However, the antibacterial and antiproliferative capacity of S. filiforme-derived products/extracts has not been characterized. Other research groups have demonstrated the nutritional value of S. filiforme to humans and animals;,, nevertheless, its cytotoxic or antimicrobial properties were not evaluated.
In this context, based on the progress recent of the Syringodium genus, this research aimed to characterize, a lipophilic fraction of S. filiforme seagrass to determine its antibacterial activity against human pathogenic micro-organisms, as well as its cytotoxic effects.
| Materials and Methods|| |
S. filiforme was collected on March 16, 2016, in Guanabo Beach (23°10'44”N - 82°07'01” W) Havana, Cuba. It was authenticated by Dr. A. J. Areces (Institute of Oceanology, Havana) and deposited in the collection of the National Aquarium of Cuba, with number IDO 165. The seagrass was washed with distilled water to remove sand and salts and then dried in an oven at a temperature of 50°C to constant weight.
Extraction and fractionation
The dried and pulverized S. filiforme leaves (200 g) were continuously macerated with 2000 mL of ethanol: H2O (1:1 v/v) over a period of 7 days at room temperature. The extract was filtered and concentrated to dryness under reduced pressure and low temperature (45°C), resulting in 2.5% of the crude hydroethanolic extract. The dry extract was transferred to a vial and placed in desiccators to remove moisture. Four grams of this extract were fractionated using mechanic agitation for 2 h with chloroform (300 mL) to obtain the non-polar fraction after filtration and drying processes, yielding 1.6%.
Analytical-grade reagents and reference patterns for gas chromatography-mass spectrometry (GC-MS) were obtained from Sigma, USA. Culture media and supplements were purchased from GIBCO (Gibco BRL, Paisley, UK).
Gas chromatography-mass spectrometry analysis
Samples of chloroform fraction (5.0 mg) from S. filiforme (SfCHCl3) were accurately weighed into a 2 mL vial, then 0.15 mL of N-methyl-N-(trimethylsilyl) trifluoroacetamide were added. Subsequently, the vial was tightly capped, heated at 80°C for 1 h and 0.5 μL were analyzed by GC-MS. The analyses were performed using an Agilent GC 6890N equipped with a mass selective detector 5975 B inert, and a split-splitless injector, in splitless mode, was used (Agilent, Palo Alto, CA, USA). Separations were made on a HP-5Ms fused-silica capillary column (30 m x 0.25 mm), with a film thickness of 0.25 μm Df (Agilent, Avondale, PA). The GC oven temperature was kept at 60°C for 2 min and programmed to 200°C at a rate of 20°C/min, then from 200°C to 300°C at a rate of 8°C/min and kept constant at 300°C for 5 min. The temperature of the injector was fixed at 320°C and that of the source at 280°C, while MS interface temperature was 250°C. Helium (purity, 99.9995%) was the carrier gas; its flow rate was fixed at 1 mL/min. Ionization of the sample components was performed in electron impact mode (EI, 70 eV). The mass range from 40 to 1000 m/z was scanned at a rate of 3.0 scans/s. One microliter of the organic extract was manually injected into the GC-MS system using a Hamilton syringe, for total ion chromatographic analysis. The total running time of the GC-MS system was 70 min. The relative percentage of each extract constituent was expressed as a percentage with respect to peak area normalization. Peak identification was achieved by computer matching mass spectra against commercial libraries (National Institute of Standards and Technology (NIST 2011 GC-MS), as well as MS literature data.,,,
Determination of antimicrobial activity
Antibacterial activity was determined by the microdilution method according to the Guidelines of National Committee of Clinical Laboratory Standards. The crude extract of S. filiforme and SfCHCl3 was assayed in concentrations between 1 and 2000 μg/mL. For this assay reference strains of Staphylococcus aureus (ATCC 6538), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 10576), Candida albicans (ATCC 10231), and Salmonella typhi (ATCC 9992 vaccine) were used. All micro-organisms were adjusted to 0.5 Mc Farland scale. Double dilutions were made from 2000 μg/mL to 1 μg/mL in 96-well plates. In each well, 25 μL of Syringodium extract or fraction plus 25 μL of each micro-organism was added. Furthermore, a positive control was performed by adding 25 μL of each micro-organism plus 25 μL of saline. Streptomycin was used as a reference antimicrobial agent. The plates were incubated at 37°C for 24 h, and then 5 μL of each dilution was plated in triplicate in Tryptone Soya Agar medium and incubated at 37°C for 24 h to determine the minimum inhibitory concentration (MIC). Reading was performed thereafter. MIC was taken as the lowest concentration of the product able to completely inhibit bacterial growth in the well. The experiment was performed in triplicate.
Cytotoxic evaluation by 3-(4,5-dimethylthiazol
-2-yl)-2,5-diphenyltetrazolium bromide reduc
Cell viability of A549 cells (ATCC CCL-185) in the presence of different concentrations of S. filiforme crude extract, SfCHCl3, and cisplatin (0.01–1000 μg/mL) were evaluated through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, as it has been previously described. The crude extract and cisplatin were dissolved in culture media, whereas the organic fraction was solubilized in culture media with dimethyl sulfoxide (DMSO) 1%. Untreated cells and cells treated with DMSO 1% were used as controls. Cell viability was determined from three independent experiments with three replicas each.
The statistical analysis was carried out using the GraphPad Prism software. Values are expressed as the mean ± standard deviation of at least three independent experiments. One way ANOVA with a Tukey posttest (P < 0.05 and P < 0.01) was used for multiple mean comparisons.
| Results|| |
The GC-MS analysis of the obtained SfCHCl3 after chemical derivatization allowed the identification of saturated and unsaturated fatty acids (FAs) (ω-3 and ω-6), phenolic acids, sterols, glycerides, fatty alcohols, and flavonoids [Figure 1] and [Table 1]. In the chromatogram, 68 components of the seventy detected were identified, being palmitic acid (C16:0) the main component (39.18%), followed by azelaic acid (5.06%), α-linolenic (C18:3) (3.63%), oleic acid (3.54%), linoleic acid (C18:2) (3.33%), and two isomers of palmitoleic acid (3.21 and 3.04%) [Table 1]. Other acidic, aromatic, and aliphatic compounds were detected in lower proportions.
|Figure 1: Gas chromatography-mass spectrometry profile of the derivatized chloroform fraction obtained from hydroethanolic extract of S. filiforme|
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|Table 1: Chemical composition of chloroform fraction from crude extract of Syringodium filiforme|
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[Table 2] shows the antibacterial activity of S. filiforme crude extract, SfCHCl3, and Streptomycin. The crude extract showed low antibacterial activity against the tested strains and in some cases, it was close to zero relative to the fraction obtained from it. Moreover, the organic fraction revealed antibacterial activity in concentrations below 1.5 mg/mL. SfCHCl3 showed good activity against strains of Staphylococcus aureus (ATCC 6538), Pseudomonas aeruginosa (ATCC 9027), and Salmonella typhi (ATCC 9992v), and it was less active against E. coli (ATCC 10576) and Candida albicans (ATCC 10231). The strains tested are not resistant to multiple drugs; however, it is very important to evaluate the inhibitory effect of natural extracts and fractions obtained from these, since they represent clinically relevant species involved in major human infections and good models to search for new antibacterial candidates. The antibacterial activity shown by this fraction led us to evaluate its cytotoxic potential in human lung cancer cells.
|Table 2: Antibacterial activity of the crude extract and chloroform fraction of Syringodium filiforme|
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The presence of long-chain FAs, as well glycerides and phenolic acids, suggests possible cytotoxic and antitumor properties for SfCHCl3. In this way, the cytotoxic potential of SfCHCl3 and S. filiforme crude extract in comparison to a clinically used anticancer drug such as cisplatin was evaluated in A549 human lung carcinoma. The organic fraction significantly reduced the viability of treated cells in comparison to control cells (no treatment or treatment with DMSO 1%), whereas the crude extract did not affect cell viability at the same concentrations [Figure 2]. SfCHCl3 exhibited concentration-dependent cytotoxicity in A549 cell line. The effects of this fraction in this cell line were comparable to the cytotoxic activity of cisplatin, which also shows a concentration-dependent effect [Figure 2].
|Figure 2: Cytotoxic activity of the chloroform fraction and the crude hydroethanolic extract of S. filiforme in A549 cells. Cell viability of A549 cells after 48 h of exposure with different concentrations of Cisplatin, the crude extract and the chloroform fraction of S. filiforme was evaluated by MTT reduction assays. The Figure shows the concentration-response curve for the cytotoxicity of the three products from 0.01 to 1000 μg/ml in comparison to untreated control cells and cells treated with dimethyl sulfoxide 1% (vehicle control). The values are shown as mean percentages of control from three independent experiments. bars, standard deviation. *P < 0.05 and ***P < 0.001 versus control (untreated cells)|
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| Discussion|| |
Sixty-eight compounds were identified in the GC-MS analysis of SfCHCl3. All of them were identified for the first time in S. filiforme that grows in Cuban coastal zones. Among the total components detected, FA resulted in the most abundant reaching a value of 70%. Accordingly, palmitic acid was the main metabolite found in S. isoetifolum.
Non-polar fractions of some species of seagrasses have been studied by GC–MS. In this sense, the analysis of chemical fractions purified from Halophila ovalis R. Br. Hooke (Hydrocharitaceae) revealed a high content of 9-octadecenoic (oleic) acid (27.01%), hexadecanoic (palmitic) acid (21.63%), and octadecanoic (stearic) acid (10.42%) in one of these fractions; while, benzoic acid (11.11%), tetradecanoic acid (6.12%), and hexadecane (3.47%) were found the prevalent components in other non-polar fractions.
Audra et al. studied the composition of non-polar fractions of three seagrasses, Halodule wrightii (shoal grass), S. filiforme, and T. testudinum, all plants analyzed contained less than 1% lipid (wet weight) and more than 75% moisture. Freshwater plants lipids were composed of monoacylglycerols, phosphatidylserine, and/or phosphatidylethanolamine. These were the major lipid classes for the seagrasses, along with phosphatidylcholine. The main FA in all plants analyzed included palmitic, linoleic, and linolenic acid, with lesser amounts of palmitoleic and oleic acid. This research demonstrated that lipid and FA distributions for seagrasses and freshwater plants, collected in local Florida waters, are similar to those found in other parts of the world. Similarly, the chemical composition of Indian seagrasses (Enhalus acoroides; Thalassia hemprichii; Halodule pinifolia; S. isoetifolium; C. serrulata and C. rotundata) were determined by GC–MS. In C. rotundata, FA was the main phytoconstituents, being palmitic acid the main metabolite. Particularly, S. isoetifolium yielded seven compounds: palmitic acid (42.88%), 9-octadecanoic acid (Z)-methyl ester (24.04%), 3, 7, 11, 15-tetramethyl-2-hexadecan-1-ol (24.04%), oleic acid (1.51%), tridecanoic acid methyl ester (1.61%), cyclopentaneundecanoic acid methyl ester (3.29%), and 13-octadecenal (3.29%).
Particularly, S. isoetifolium yielded seven compounds: palmitic acid (42.88%), Methyl octadecenoate (24.04%), 3, 7, 11, 15-tetramethyl-2-hexadecan-1-ol (24.04%), oleic acid (1.51%), methyl tridecanoate (1.61%), methyl cyclopentaneundecanoate (3.29%), and 13-octadecenal (3.29%).
It is interesting to point out the presence of a high concentration (5%) of azelaic acid in S. filiforme. This saturated dicarboxylic acid is reported to be produced by the symbiotic fungus Pityrosporum ovale present in human skin and commonly found in terrestrial plants such as Hordeum vulgare (barley), Secale cereale (rye), some species of Triticum genus (wheat) and in some well-characterized angiosperms such as Arabidopsis thaliana,,,, but it is not abundant in marine organisms. Azelaic acid has been detected in marine aerosols, as a result of photooxidation of FAs produced by phytoplankton, and there is one reference of a 3% content of this acid in the same studied fractions of the macroalgae Ulva lactuca, and in the marine species T. testudinum but in lower concentration (1%).
Other compounds with around 7% of contribution in this chloroform fraction were hydrocarbons, being tricosane (1.93%), tetracosane (1.81%), pentacosane (1.45%), and heptacosane (1.53%) the most abundant. Some of them have also been detected in other seagrass genera such as Halodule, Halophila, and Thalassia.,,, Minor sterols such as sitosterol were found in our study of S. filiforme species, which is in accordance with those reported in S. isoetifolium and other marine angiosperms of Cymodocea, Halodule, Thalassia, and Enhalus genera from tropical zones after GC-MS analysis.,,
On the other hand, phenolic acids derived from benzoic acid were detected. They include ρ-hydroxybenzoic as a major component (2.57%), ρ-hydroxy-3-methoxybenzoic acid (vanillic acid); 3,5-dihydroxy benzoic acid methyl ester; 3,4-dihydroxybenzoic acid, and the benzoic acid itself. It is well known the pharmacologic potential of these compounds, which are also present in the abovementioned marine angiosperms.
Plants are important sources of naturally occurring antimicrobial and anticancer agents. Antimicrobial and cytotoxic activities of plant extracts have been extensively reported in literature.,,, Some of these studies have led to the identification of the active components responsible for such activities, contributing to the development of novel drugs for therapeutic use in humans. Because of the emergence of multiple drug resistance in human pathogenic micro-organisms and the adverse effects of cancer chemotherapeutic drugs, the search for new antimicrobial and anticancer molecules from alternative sources, including plants, is receiving attention by the scientific community.,,,
Seagrasses are continuously defending from the infection of marine micro-organisms. In this way, many seagrass species are able to produce secondary metabolites with antibacterial and antifungal properties. For example, Engel et al. showed the antimicrobial activity of a lipophilic fraction from S. filiforme against S. aggregatum and P. bacteriolytica strains that can affect marine plants. By a similar approach, Yuvaraj et al. highlighted the presence of the same activity in H. ovalis extracts, which proved effective against several bacteria and Vibrio. T. hemprichii flavonoids also possessing in vitro capabilities of inhibiting different bacteria. Similarly, Aswathi et al. demonstratedsignificant antibacterial activity of acetone and methanol extracts from S. isoetifolium against Gram stains of human and fish pathogens, proving a mayor antimicrobial effect for the acetone extract. In addition, Iyapparaj et al. have also showed the antibacterial and antifouling capacity of acetone, dichloromethane, and methanol extracts from S. isoetifolium and other marine plants such as C. serrulata.
In the present study, SfCHCl3 exhibited antimicrobial activity against five human pathogens, particularly against S. aureus, P. aeruginosa, and S. typhi; whereas S. filiforme crude extract displayed lower effects and was no active against two of the five tested strains. This difference could be related to the high content of FAs in the fraction, since they have been reported to mediate the antibacterial effects of many organic fractions and extracts from marine plants and seaweeds.,, Furthermore, azelaic acid has been described as an antimicrobial agent that inhibits the proliferation of food pathogenic bacteria; it is indicated for the treatment of acne rosacea and cutaneous infections and has been proven to be well tolerated in numerous clinical trials. This substance has also shown profound anti-inflammatory, antioxidant, and cytotoxic effects, thus being another compound that could synergistically contribute to the observed effects of SfCHCl3.
Cytotoxicity is an interesting pharmacological property that has also been studied for different extracts and pure compounds isolated from seagrasses. For instance, a significant in vitro cytotoxic effects against two human lung cancer cell lines were demonstrated by cymodienol from Cymodocea nodosa. In contrast, ketosteroids from C. nodosa showed no in vitro cytotoxicity against these cell lines. Other compounds, such as the syphonoside from Halophila stipulacea, showed no cytotoxicity, however, inhibited apoptosis in some of the studied cell lines. On the other hand, El Baz et al. demonstrated antitumor activity of sulfolipid fractions, mainly composed for FA derivatives, from various algal species against HepG2 and MCF-7 cell lines, as well as antibacterial activity against B. subtilis and E. coli. Both activities found are attributable to those kinds of compounds. Similarly, crude extracts of the marine plant Thalassodendron ciliatum exhibited cytotoxic effects against selected human cancer cell lines, and some activity against hepatitis A and herpes simplex viruses in vitro.,
According to these previous evidence and based on the preferential antimicrobial activity that SfCHCl3 displays compared with the crude extract, we decided to evaluate the cytotoxicity of the lipophilic fraction in human lung cancer cells, in comparison to S. filiforme crude extract and a reference chemotherapeutic agent used for lung cancer treatment, such as cisplatin. This study revealed important cytotoxic effects of SfCHCl3 in the cell line A549, which were similar to the cytotoxicity of cisplatin in the same cell line. As it occurred with the antimicrobial activity, the crude extract was no cytotoxic in comparison with the chloroform fraction and was not able to decrease the viability of A549 cells below an 80%, not even at the highest concentration tested.
The main compound present in SfCHCl3 was palmitic acid. This FA showed cytotoxic effects in A549 human lung cancer cell line through a mechanism that involves endoplasmic reticulum stress, hypercalcemia, and generation of reactive oxygen species in the study carried out by Wong et al. Furthermore, other lipophilic mixtures, containing short and long-chain FA exhibit cytotoxic effects in different human cancer cell lines and enhance the activity of cytostatic drugs such as paclitaxel and cisplatin., Because of that, some of these acids and their methyl esters have also drawn the attention and have been commercialized as nutritional supplements showing antioxidant, anticancer, and antihistaminic properties.,
Recently, our group carried out a similar procedure to study T. testudinum leaves from Cuban coastal zones and it was found that the same fraction was mainly composed of FAs (80%), where palmitic acid was also the major compound. Similarly, such studied fraction showed a potent cytotoxicity and antiproliferative effect in the same cell lines, which is in agreement with the results obtained in the present study.
As previously mentioned, azelaic acid could be contributing to the cytotoxicity of SfCHCl3. Besides its antimicrobial properties, this compound has demonstrated cytotoxic and antiproliferative effects.,, At molecular level, azelaic acid acts as a competitive inhibitor of tyrosinase, a key enzyme for melanin synthesis; consequently, it has been used to treat hyperpigmented disorders including melasma and lentigo maligna,,, but it has also shown antitumor activity on melanoma, leukemia, and squamous carcinoma cells in vitro and in vivo in mice with xenotransplanted melanoma tumors.,, This dicarboxilic acid has been tested in humans as well, demonstrating effectiveness against melanoma in situ and malignant melanoma.
The present work reveals the importance of non-polar and mid-polar constituents of Syringodium filiforme for its pharmacological properties. Since we demonstrate that the crude extract of S. filiforme, which is predominantly polar, is not active compared with the chloroform fraction isolated from it, our results indicate that the non-polar components present in S. filiforme are critical for the biological activity of products derived from this marine plant. Based on the presented evidence, it can be inferred that the antibacterial and cytotoxic effects demonstrated for SfCHCl3 may be due to the presence of FAs and azelaic acid, in synergy with other minor components of the fraction such as glycerides and flavonoids. This work contributes to the chemo-taxonomical characterization of S. filiforme and validates the chloroform fraction derived from the crude extract of this organism as a potential source of natural antimicrobial and cytotoxic molecules.
| Conclusion|| |
The lipidic extract of the phanerogam Syringodium filiforme was characterized by GC-MS. The assays allowed the identification of the main non-polar compounds of this species and, for the first time, demonstrated its antimicrobial effects against human pathogens and cytotoxicity in A549 human lung carcinoma. On the basis of these facts, this seagrass is an interesting potential biotechnological resource.
Financial support and sponsorship
This work has been supported by the research project: “Conservation and sustainable management of natural resources in Rincon de Guanabo Protected Natural Landscape for the benefit of the community” of UNDP Small Donation Fund in Cuba and the project “Uses and benefits of marine plant Thalassia testudinum” (P211LH005-015) from the Environment Agency, Cuban Ministry of Science, Technology, and Environment (CITMA). The authors also which to acknowledge Cuban diver Jose Ramon Garcia for collecting Syringodium filiforme samples.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Berdy J. Bioactive microbial metabolites. J Antibiot 2005;58:1-26.
Mishra BB, Tiwari VK. Natural products: An evolving role in future drug discovery. Eur J Med Chem 2011;46:4769-807.
Molinski T, Dalisay D, Lievens, P
Saludes J. Drug development from marine natural products. Nat Rev Drug Discov 2009;8:69-85.
Sudek S, Lopanik NB, Waggoner LE, Hildebrand M, Anderson C, Liu H, Patel A, et al
. Identification of the putative bryostatin polyketide synthase gene cluster from Candidatus Endobugula sertula, the uncultivated microbial symbiont of the marine bryozoan Bugula neritina. J Nat Prod 2007;70:67-74.
Schumacher M, Kelkel M, Dicato M, Diederich M. Gold from the sea: Marine compounds as inhibitors of the hallmarks of cancer. Biotechnol Adv 2011;29:531-47.
Williams DH, Stone MJ, Hauck PR, Rahman SK. Why are secondary metabolites (natural products) biosynthesized? J Nat Prod 1989;52:1189-208.
Firn RD, Jones CG. Natural products: A simple model to explain chemical diversity. Nat Prod Rep 2003;20:382-91.
Simmons TL, Andrianasolo E, McPhail K, Flatt P, Gerwick HW. Marine natural products as anticancer drugs. Mol Cancer Ther 2005;4:333-42.
Puglisi MP, Engel S, Jensen PR, Fenical W. Antimicrobial activities of extracts from Indo-Pacific marine plants against marine pathogens and saprophytes. Mar Biol 2007;150:531-54.
Ragupathi RK, Arumugam R, Anantharama P. Chemical composition and antibacterial activity of Indian seagrasses against urinary tract pathogens. Food Chem 2012;135:2470-73.
Zidorn C. Secondary metabolites of seagrasses (Alismatales and Potamogetonales; Alismatidae): Chemical diversity, bioactivity and ecological function. Phytochemistry 2016;124:5-28.
Riera M, Marrero D, Hernandez I, González K, Pérez D, Manso A, Labrada M, et al
. Chemical characterization and cytotoxic potential of a chloroform fraction obtained from marine plant thalassia testudinum. J Chromatogr Sep Tech 2018;9:1-6.
Athiperumalsami T, Rajeswari VD, Poorna SH, Kumar V, Jesudass LL. Antioxidant activity of seagrasses and seaweeds. Bot Mar 2010;53:251-7.
Pushpa BN, Amudha P, Varadharaj V. Sea grasses Novel marine nutraceuticals. IJPBS 2016;7:567-73.
Ragupathi RK, Arumugam R, Anantharaman P. In vitro
antibacterial, cytotoxicity and haemolytic activities and phytochemical analysis of seagrasses from the gulf of mannar, South India. Food Chem 2013;136:1484-9.
Nuissier G, Rezzonico B, Dubois GM. Chicoric acid from Syringodium filiforme. Food Chem 2010;120:783-8.
Nuissier G, Diaba F, Dubois GM. Bioactive agents from beach waste: Syringodium flotsam evaluation as a new source of L-chiro-inositol. Innov
Food Sci Emerg 2008;9:396-400.
González KL, Valdés O, Laguna A, Díaz M, González LJ. Antioxidant effect and polyphenol content of Syringodium filiforme
(Cymodoceaceae). Rev Biol Trop 2011;59:465-72.
Aswathi EM, Velammal A, Patterson j phytochemicals of the seagrass syringodium isoetifolium and its antibacterial and insecticidal activities. EJBS 2012;4:63-7.
Engel S, Puglisi M, Jensen P. Fenical W. Antimicrobial activities of extracts from tropical Atlantic marine plants against marine pathogens and saprophytes. Mar Biol 2006;149:991-1002.
Hamdy A, Metwally W, El Fotouh M, Rodríguez B, El Dewany A, El Toumy S, et al.
Bioactive phenolic compounds from egyptian red sea seagrass thalassodendronh ciliatum. Z. Naturforsch C 2010;67:291-6.
Rodeiro I, Hernandez I, Herrera JA, Romo RM, Donato MT, Tolosa L, et al
. Assessment of the cytotoxic potential of an aqueous-ethanolic extract from Thalassia testudinum angiosperm marine grown in the Caribbean Sea. J Pharm Pharmacol 2018;70:1553-60.
Iyapparaj P, Revathi P, Ramasubburayan R, Prakash S, Palavesam A, Immanuel G, et al
. Antifouling and toxic properties of the bioactive metabolites from the seagrasses syringodium isoetifolium and cymodocea serrulata. Ecotox Environ Saf 2014;103:54-60.
Coria E, Durán E. Proximal analysis of seagrass species from laguna de términos, mexico. Hidrobiológica 2015;25:249-55.
Coria E, Duran E. The relationship between the massive nesting of the olive ridley sea turtle (Lepidochelys olivacea) and the local physical environment at la escobilla, oaxaca, mexico, during 2005. Hidrobiológica 2017;27:201-09.
Siegal JL, Harr K, Lee CH, Scott KC, Gerlach T, Reuter M, et al
. Proximate nutrient analyses of four species of submerged aquatic vegetation consumed by florida manatee (trichechus manatus latirostris) compared to romaine lettuce (lactuca sativa var. longifolia). J Zoo Wildl Med 2010;41:594-602.
Regalado EL, Rodriguez M, Menéndez R, Fernández X, Hernández I, Morales RA, et al
. Photoprotecting action and phytochemical analysis of a multiple radical scavenger lipophilic fraction obtained from the leaf of the seagrass thalassia testudinum. Photochem Photobiol 2011;87:1058-66.
Rodeiro I, Olguín S, Santes R, Herrera JA, Pérez CL, Mangas R, et al
. Gas chromatography-mass spectrometry analysis of ulva fasciata (green seaweed) extract and evaluation of its cytoprotective and antigenotoxic effects. Evid-Based Compl Alt 2015;2015:1-11.
Valle H, Ospina S, Galeano E, Martínez A, Marquez ME, Lopéz JB. Chemical compounds of antimitotic fraction in ethanol extract from seaweed Digenia simplex. Revista de la facultad de química farmacéutica. Universidad de Antioquia, Medellín, Colombia 2008;15:141-49.
Wayne PA. National Committee for Clinical Laboratory Standards. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically; 2000.
Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
Yuvaraj N, Kanmani P, Satishkumar R, Paari A, Pattukumar V, Arul V. Seagrass as a potential source of natural antioxidant and anti-inflammatory agents. Pharm Biol 2012;50:458-67.
Audra LA, Van Vleet ES, Clinton D. Characterization of lipids in aquatic vegetation from the west coast of Florida. Fla Sci 2007;70:264-74.
Parvez S, Kang M, Chung HS, Bae H. Naturally occurring tyrosinase inhibitors: Mechanism and applications in skin health, cosmetics and agriculture industries. Phytother Res 2007;21:805-16.
Zouboulis CC, Katsambas AD, Kligman AM, Editors. Pathogenesis and Treatment of Acne and Rosacea Springer. Berlin: Heidelberg; 2014.
Pusztahelyi T, Holb IJ, Pócsi I. Secondary metabolites in fungus-plant interactions. Front. Plant Sci 2015;6:573.
Van Alfen NK, Editor. Encyclopedia of Agriculture and Food Systems. Academic Press. Elsevier; 2014.
Mochida M, Kitamori Y, Kawamura K, Nojiri Y, Suzuki K. Fatty acids in the marine atmosphere: Factors governing their concentrations and evaluation of organic films on sea salt particles. J Geophys Res 2002;107:4325-34.
Attaway DH, Parker PL, Mears JA. Normal Alkanes of Five Coastal Spermatophytes, Publications of the Institute of Marine Science. Vol. 15. University of Texas; 1970. p. 13-9.
Jerković I, Marijanović Z, Roje M, Kuś PM, Jokić S, Rakovac CR. Phytochemical study of the headspace volatile organic compounds of fresh algae and seagrass from the Adriatic Sea (single point collection). PLoS ONE 2018;13:e0196462.
Gillan FT, Hogg RW, Drew EA. The sterol and fatty acid compositions of seven tropical seagrasses from north queensland, Australia. Phytochemistry 1984;23:2817-21.
Govindan M, Hodge JD, Brown KA, Smith NM. Distribution of cholesterol in Caribbean marine algae. Steroid 1993;58:178-80.
Al-Hadhrami and Hossain. Evaluation of antioxidant, antimicrobial and cytotoxic activities of seed crude extracts of Ammi majus grown in Oman. EJBAS 2016;3:329-34.
Kumar S, Chashoo G, Saxena AK, Pandey AK. Parthenium hysterophorus: A probable source of anticancer, antioxidant and anti-HIV agent. Bio Med Res Int 2013;2013:1-11.
Kumar S, Pandey S, Pandey AK. In vitro
antibacterial, antioxidant and cytotoxic activities of Parthenium hysterophorus and characterization of extracts by LC-MS analysis. J Biomed Biotechnol 2014;2014:1-10.
Rahman A, Islam S. Antioxidant, antibacterial and cytotoxic effects of the phytochemicals of whole Leucas aspera extract. Asian Pac J Trop Biomed 2013;3:273-79.
Kumar S, Bhargava A, Sharma B, Pandey AK. Studies on in vitro
antioxidant and antistaphylococcal activities of some important medicinal plants. Cell Mol Biol 2011;57:16-25.
Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. The Sci World J 2013;2013:1-16.
Martin KW and Ernst E. Herbal medicines for treatment of bacterial infections: A review of controlled clinical trials. J Antimicrob Chemother 2003;51:241-6.
Suffredini IB, Sader HS, Gonçalves AG, Reis AO, Gales AC, Varella AD, et al
. Screening of antibacterial active extracts obtained from plants native to Brazilian amazon rain forest and atlantic forest. Brazilian J Med Biol Res 2004;37:379-84.
Maximilien R, De Nys R, Holmstrom C, Gram L, Givskov M, Crass K, et al
. Chemical mediations of bacterial surface colonization by secondary metabolites from the red alga Delisea pulchra. Aquat Microb Ecol 1998;15:233-24.
Hawas UW. A new 8-hydroxy flavone o-xyloside sulfate and antibacterial activity from egyptian seagrass thalassia hemprichii. Chem Nat Comp 2014;50:629-32.
Ravikumar S, Nanthini K, Ajith KT, Ajmalkhan M. Antibacterial activity of seagrass species of cymodocea serrulata against chosen bacterial fish pathogens. Ann Biol Res 2011;2:88-93.
Shanmughapriya S, Manilal A, Sugathan S, Selvin J, Kiran S, Natarajaseenivasan K. Antimicrobial activity of seaweeds extracts against multiresistant pathogens. Ann Microbiol 2008;58:535-41.
Moniharapon T, Moniharapon E, Watanabe Y, Hashinaga F. Inhibition of food pathogenic bacteria by azelaic acid. PJBS 2005;8:450-55.
Sieber MA, Hegel JKE. Azelaic Acid: Properties and mode of action skin. Pharmacol Physiol 2014;27:9-17.
Konitza I, Vagias C, Jakupovic J, Moreau D, Roussakis C, Roussis V. Cymodienol and cymodiene: New cytotoxic diarylheptanoids from seagrass Cymodosea nodosa. Tetrahedron Lett 2005;46:2845-47.
Kontiza I, Abatis D, Malakate K, Vagias C, Roussis V. 3-Keto steroids from the marine organisms dendrophyllia cornigera and cymodosea nodosa. Steroids 2006;71:177-181.
Gavagning M, Carbone M, Amodeo P, Mollo E, Vitale RM, Roussis V, Cimino G. Structure and absolute stereochemistry of syphonoside, a unique macrocyclic glicoterpenoidfrom marine organism. J Org Chem 2007;72:5625-30.
El Baz FK, El Baroty GS, Abd El Baky HH, Abd El-Salam OI, Ibrahim EA. Structural characterization and biological activity of sulfolipids from selected marine algae. Grasas Aceites 2013;64:561-71.
Hamdy A, Mettwally W, Fotouh M, Rodriguez B, Dewany A, El-Toumy S, et al
. Bioactive phenolic compounds from the egyptian red sea. Z. Naturforsch 2012;67c: 291-96.
Wong KL, Cheung C, Cheung So E, Huang BM, Leung YM. Palmitic acid-induced cytotoxicity in human alveolar A549 cells involved endoplasmic reticulum (Er) stress and reactive oxygen species production. Anesth Analg 2016;123:386.
Menéndez JA, Ropero S, Mar del Barbacid M, Montero S, Solanas M, Escrich E, et al
. Synergistic interaction between vinorelbine and gamma-linolenic acid in breast cancer cells. Breast cancer research and treatment. Breast Cancer Res Tr 2002;72:203-19.
Zajdel A, Wilczok A, Latocha M, Tarkowski M, Kokoci-Ska M. Polyunsaturated fatty acids potentiate cytotoxicity of cisplatin in A549 cells. Acta Pol Pharm 2014;71:1060-65.
Melariri P, Campbell W, Etusim P, Smith P. In vitro
and in vivo
antimalarial activity of linolenic and linoleic acids and their methyl esters. Adv Stud Biol 2012;4:333-49.
Fitton A, Goa KL. Azelaic acid. A review of its pharmacological properties and therapeutic efficacy in acne and hyperpigmentary skin disorders. Drugs 1991;41:780-98.
Breathnach AS. Azelaic acid: Potential as a general antitumoural agent. Med Hypotheses 1999;52:221-26.
Sun J, Keun A. Effect of combination of taurine and azelaic acid on antimelanogenesis in murine melanoma cells. J Biomed Sci 2010;17:S45.
[Figure 1], [Figure 2]
[Table 1], [Table 2]