Manipulation of fatty acid profile and nutritional quality of Chlorella vulgaris by supplementing with citrus peel fatty acid.

Microalgae could be an excellent resource of functional and essential fatty acids. To achieve viable microalgal biomass production, mass cultivation of microalgae is required; however, the high cost of nutrients is the obstacle. An inexpensive and nutritious material is required to feed Chlorella vulgaris in the pharmaceutical and food sectors. Citrus peel waste with a valuable nutritional quality could be one of the promising and inexpensive candidates. In this study, the fatty acid extract from different citrus peels was used as the organic nutrient source for the cultivation of Chlorella. The proximate composition of bitter orange, sweet orange, grapefruit, and mandarin peels were determined, and their nutritional quality was evaluated. Total fatty acids from the citrus peel were prepared by acidic methanol hydrolysis and hexane extraction. Fourier transforms infrared (FT-IR) and gas chromatography-mass spectrometry (GC-MS) was used to analyze the fatty acid composition and nutrient composition. Fatty acids from the citrus peels were added to the Chlorella culture medium to study their influences on biomass, lipid production, fatty acid profile, and nutritional quality of Chlorella. The most predominant citrus peel fatty acids were linoleic, palmitic, oleic, linolenic, and stearic acids. The citrus peels contain polyunsaturated, saturated, and monounsaturated fatty acids. The most unsaturated fatty acids were omega-6, omega-3, omega-9, and omega-7. The citrus peel had acceptable atherogenicity, thrombogenicity, omega-6/omega-3, peroxidizability, hypocholesterolemic, and nutritive value indices. The major fatty acids of Chlorella were palmitic, linoleic, oleic, alpha-linolenic, gamma-linolenic, 4,7,10,13-hexadecatetraenoic, palmitoleic, 7,10-hexadecadienoic, 7,10,13-hexadecatrienoic, lauric and 5,8,11,14,17-eicosapentaenoic acids. Chlorella contains polyunsaturated, saturated, and monounsaturated fatty acids. The most unsaturated fatty acids contain omega-6, omega-3, omega-9, and omega-7. Chlorella had acceptable atherogenicity, thrombogenicity, omega-6/omega-3, hypocholesterolemic, peroxidizability, and nutritive value indices. Supplementation of Chlorella with citrus peels fatty acid increases total biomass, lipid content, and nutritional quality of Chlorella. The present research shows that citrus peels have good nutritional quality and could be used for the inexpensive cultivation of Chlorella biomass with potential utility for food application.

Anti-oxidative and anti-hyperglycemic properties of Agastache foeniculum essential oil and oily fraction in hyperglycemia-stimulated and lipopolysaccharide-stimulated macrophage cells: In vitro and in silico studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Hyperglycemia (HG) and lipopolysaccharide (LPS) often promote superoxide accumulation, which may increase oxidative stress. Reducing superoxide production in hyperglycemia and the inflammatory condition is an emerging way to reduce protein and lipid oxidation and diabetes complication. AIM OF STUDY: To examine the effect of Agastache foeniculum essential oil (AFEO) and oil fraction (AFoil) on HG- and LPS-stimulated oxidative stress, the pathogenicity of AFEO and AFoil on oxidative stress was assessed. METHODS: The stimulatory effects of AFEO and AFoil on the activity and expression of NADH oxide (NOX), catalase (CAT), superoxide dismutase (SOD), and the expression of nuclear respiratory factor 2 (NRF2) and nuclear factor-kappa B (NF-kB) in the stimulated macrophage cell line, J774.A1, was studied. The interaction patterns of AFEO and AFoil components with NOX, SOD, CAT, NRF2, and NF-kB proteins were also deduced using molecular docking. RESULTS: Estragole was the main ingredient in AFEO (97%). Linolenic acid (32.10%), estragole (16.22%), palmitic acid (12.62%), linoleic acid (12.04%), and oleic acid (8.73%) were the major chemical components of the AFoil. NOX activation was stimulated in macrophage cells by HG and LPS. At 20 µg/mL, AFEO and AFoil decreased NOX activity while increased SOD and CAT activities in stimulated macrophages. AFoil with estragole and omega-3 fatty acids was better than AFEO with estragole in anti-hyperglycemic and anti-oxidative activity. According to molecular docking research, estragole, linoleic acid, and linolenic acid bind to different hydrophobic pockets of NOX, SOD, CAT, NFR2, and NF-kB using hydrogen bonds, van der Waals bonds, pi-alkyl, and pi-anion interactions, with different binding energies. CONCLUSION: AFEO and AFoil showed antioxidant and anti-diabetic activity. The mechanisms in lowering oxidative stress markers depended on down-regulating superoxide-producing enzymes and up-regulating superoxide-removing enzymes at gene and protein levels. The AFoil emulsion can be used to reduce the detrimental impacts of hyperglycemia and oxidative stress.

Synergistic effect of Zataria Multiflora essential oil on doxorubicin-induced growth inhibition of PC3 cancer cells and apoptosis.

Today, herbs are used as adjuncts to reduce the toxicity of chemotherapy drugs. Here, Zataria-Multiflora Essential Oil (ZEO) was concomitantly employed with doxorubicin, as an anti-cancer drug, to reduce the doxorubicin dosage. The growth inhibition was determined using MTT assay in treated cells. The morphological alteration was observed by fluorescent staining. To verify and compare the apoptosis, AnnexinV-PI flowcytometry and DNA fragmentation assay were performed, and the influence of the compounds on ROS generation was assessed. Changes in MMP and protein expression were analyzed by flowcytometry and western blot, respectively. The results showed that ZEO can act as an amplifier to sensitize PC3 prostate cancer cells to undergo ROS generation and apoptosis. This amplification can heighten the doxorubicin efficacy in lower doses. Consequently, our results indicated that doxorubicin-ZEO combinatory treatment of PC3 cells can reduce the nonspecific toxicity of doxorubicin and can be considered as a candidate in combinatory therapy.

Development of pre-emergence herbicide based on Arabic gum-gelatin, apple pectin and savory essential oil nano-particles: A potential green alternative to metribuzin.

This study was conducted as a plot experiment to investigate the phytotoxicity effects of nano-encapsulated savory essential oil (EO) when it is incorporated separately into carbohydrate and protein natural polymers (Arabic gum-gelatin, apple pectin and gelatin) and two cross-linkers including one poly acid and one enzyme (citric acid and transglutaminase enzyme). Each product was tested as a pre-emergence herbicide against amaranth and tomato. The evaluations also involved determining the stability, morphology, encapsulation efficiency and release properties of the prepared formulations. Coating the savory EO with cross-linked biopolymers enhanced its stability and herbicidal activity, compared to the EO nano-emulsion without any polymer or cross-linker. Among the tested formulations, the strongest inhibitory effect against amaranth germination and growth was caused by Arabic gum-gelatin and apple pectin biopolymers at the concentration of 3 ml/L of EO, when cross-linked with citric acid. These two treatments had slight effects on tomato seedlings, however. The suppressive ability of the formulations was almost similar and comparable to the chemical herbicide metribuzin (1.75 g/L). In conclusion, Arabic gum-gelatin and apple pectin cross-linked by citric acid containing savory EO can be considered as potential, green and safe replacements for metribuzin in organic tomato production.

Stabilization of Zataria essential oil with pectin-based nanoemulsion for enhanced cytotoxicity in monolayer and spheroid drug-resistant breast cancer cell cultures and deciphering its binding mode with gDNA.

Efficacy of chemotherapy is limited by the resistance of cancer cells. Phytochemicals especially Essential Oils (EOs) provide an alternative mode of cancer therapy. However, EOs utilization is restricted because of low bioavailability, and high degradation. Nanoemulsification is a method developed to overcome these obstacles. Accordingly, Citrus-Pectin nanoemulsion of Zataria Essential Oil (CP/ZEONE) was prepared to evaluate the anticancer activity and the mechanisms responsible for the caused cytotoxicity. Physical properties and FTIR spectra of CP/ZEONE were characterized. CP/ZEONE progressively improves the suppression of viability of drug-resistant MCF-7, MDA-MB-231 breast cancer cells, and spheroids. It triggers apoptosis by increasing Reactive Oxygen Species (ROS), mitochondrial membrane potential (MMP) loss, DNA damage, G2 and S-phase arrest in MDA-MB-231 cells and spheroids respectively. Additionally, spectroscopy techniques revealed the interaction of CP/ZEONE with DNA via the formation of a groove binding/partial intercalative complex. Thus, ZEO-loaded CP Nano-particles can be further explored as a promising antiproliferative and therapeutic candidate against cancer.

In-vitro (2D and 3D cultures) and in-vivo cytotoxic properties of Zataria multiflora essential oil (ZEO) emulsion in breast and cervical cancer cells along with the investigation of immunomodulatory potential.

ETHNOPHARMACOLOGICAL RELEVANCE: Zataria multiflora is an iranian valuable traditional plants, called Avishan Shirazi in Persian language used to reduce inflammation, spasm, pain, and cancer symptoms. Zataria essential oil (ZEO) is one of the essential oils possessing broad biological activities. AIM OF THE STUDY: The aim was to investigate the anticancer effects of ZEO both in-vitro and in-vivo using mouse mammary carcinoma 4T1 cell line and mouse cervical cancer TC1 cell line. MATERIAL AND METHODS: The in-vitro effects of ZEO on the proliferation of these cell lines were considered in 2D and 3D culture by MTT assay. In the following, to indicate death mode, fluorescence staining, AnnexinV/PI flowcytometry and caspase-3 activity assay of monolayer cells treated with ZEO was done. In order to evaluate the antitumor activities of ZEO, tumor-bearing BALB/c and C57BL/6 mice were intraperitoneally administered with ZEO and the immunomodulatory effects of ZEO were considered through cytokine assay. Additionally, hematobiochemical factors including aspartate aminotransferase and alanine aminotransferase were investigated to confirm the harmless effects of ZEO. RESULTS: The In-vitro results showed that treatment of cells with ZEO leads to significant inhibition of 4T1 and TC1 cell proliferation and apoptosis in monolayer cell culture (2D) and multicellular spheroids (3D). Based on In-vivo results, ZEO was effective in decreasing the tumor weight compared to the control. Furthermore, ZEO was effective in tilting the balance of cytokines in favor of T helper 1 through the increase in the secretion of TNF-α, IFN-γ, IL-2 and decrease in IL-4. During the treatment with ZEO, hematobiochemical factors of mice did not significantly change. CONCLUSION: the present study demonstrated that the ZEO has potent antiproliferative, apoptosis-inducing and immune system stimulant properties in breast and cervical cancer.

In-vitro and in-vivo anti-breast cancer activity of OEO (Oliveria decumbens vent essential oil) through promoting the apoptosis and immunomodulatory effects.

ETHNOPHARMACOLOGICAL RELEVANCE: Oliveria decumbens vent is a valuable plant in Iran, used as a vegetable. Traditionally, the aerial parts of this plant are used to treat the cancer-related symptoms, inflammation, pain, and feverish conditions. However, the scientific evidence related to its traditional effects especially the possible cellular and molecular mechanisms needs to be illuminated. AIM OF THE STUDY: The main objectives of our study were to explore in-vitro anti-cancer properties of OEO in 2D and 3D conditions, to understand the mechanism of OEO in the induction of death in cancer cells, and to identify in-vivo anti-tumor effect of OEO and induced immunomodulatory effects. MATERIAL AND METHODS: OEO was extracted by hydrodistillation and analyzed by GC-MS method. To evaluate the cytotoxic effect of OEO on 4T1 cancer monolayer cells (2D culture) and spheroids (3D cultures), MTT (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay was used. Fluorescence staining, various flow cytometry techniques, colorimetric assay, electrophoresis, and comet assay were performed to understand the anti-cancer mechanisms of OEO and determine the death mode in treated 4T1 cells. In animal studies, mouse mammary tumor model was established, the anti-tumor effect of OEO was investigated and ultimately by using the ELISA cytokine assay, immunostimulatory of OEO was studied. RESULTS: According to GC/MS analysis, thymol, carvacrol, p-cymene, and γ-terpinene were identified as main components of OEO. Based on MTT assay, OEO inhibited viability in 4T1 cancer cell without any significant effect on L929 normal cells in 2D, also the anti-proliferative effects of OEO on 4T1 spheroids (3D) was significant but less extent. Our results revealed that OEO induces apoptosis through ROS generation, mitochondrial membrane potential (ΔΨm) disruption, caspase3 activation, and DNA damage. Evaluating the effectiveness of OEO on 4T1 tumor-challenging mice and cytokine assay confirmed anti-tumor effects of OEO and development of an immune response related to Th1 expansion. CONCLUSION: These data shed light on the apoptotic mechanisms related to OEO cytotoxicity and introduced this compound as a candidate in cancer therapy.

In-vitro evaluation of apoptotic effect of OEO and thymol in 2D and 3D cell cultures and the study of their interaction mode with DNA.

Oliveria decumbens is an Iranian endemic plant used extensively in traditional medicine. Recently, some studies have been performed on biological effects of Oliveria essential oil (OEO). However, to our knowledge, the anticancer activity of OEO has not been reported. Based on our GC/MS analysis, the basic ingredients of OEO are thymol, carvacrol, p-cymene and γ-terpinene. Therefore, we used OEO and its main component, thymol, to explore their effects on cell growth inhibition and anticancer activity. Despite having a limited effect on L929 normal cells, OEO/thymol induced cytotoxicity in MDA-MB231 breast cancer monolayers (2D) and to a lesser extent in MDA-MB231 spheroids (3D). Flow cytometry, caspase-3 activity assay in treated monolayers/spheroids and also fluorescence staining and DNA fragmentation in treated monolayers demonstrated apoptotic death mode. Indeed, OEO/thymol increased the Reactive Oxygen Species (ROS) level leading to mitochondrial membrane potential (MMP, ΔΨm) loss, caspase-3 activation and DNA damage caused S-phase cell cycle arrest. Furthermore, immunoblotting studies revealed the activation of intrinsic and maybe extrinsic apoptosis pathways by OEO/thymol. Additionally, in-vitro experiments, indicated that OEO/thymol interacts with DNA via minor grooves confirmed by docking method. Altogether, our reports underlined the potential of OEO to be considered as a new candidate for cancer therapy.

Oxidative DNA damage induced by ROS-modulating agents with the ability to target DNA: A comparison of the biological characteristics of citrus pectin and apple pectin.

DNA targeting anticancer agents have been very successful in clinic, especially, when used in combinatorial therapy. But unfortunately, they often exhibit high levels of toxicity towards normal cells. Hence, much effort has been put into finding agents with more selectivity, and less toxicity. Pectins are natural polysaccharides, and beneficial nutritional fibers that have attracted attentions due to their antitumor properties. However, their molecular targets, and mechanism of action are widely unknown. Here, we have reported that citrus pectin (CP) and apple pectin (AP) selectively suppress viability in MDA-MB-231, MCF-7 and T47D human Breast cancer cells, while non-toxic to L929 normal cells. Upon CP, and AP treatments, cancer cells' ROS content increased rapidly, and led to the collapse of the mitochondrial transmembrane potential which functions upstream of the caspase-dependent apoptosis. CP and AP treated cancer cells were also arrested at the S and G1 or G2/M phases of the cell cycle, respectively. Furthermore, mRNA expression of Galectin-3 (a multi-functional lectin involved in cell adhesion, cell cycle, and apoptosis) reduced in both CP and AP treated cells. Growth inhibition of MDA-MB-231 cells by CP, and AP was concomitant with DNA damage (oxidation, and strand breaks). In this context, in an effort to clarify the mechanism of action, we showed that CP, and AP are able to interact with DNA. The strength and mode of DNA binding were established by spectroscopy techniques. We demonstrated that CP, and AP bind to dsDNA by intercalation, and groove binding/partial intercalation, respectively. In conclusion, our findings suggest that CP, and AP induce apoptosis in MDA-MB-231 cells by increasing the release of ROS, which may be related to the mitochondrial apoptosis pathway, and direct interactions with DNA. Our data indicate that these compounds may be potentially useful in cancer treatment.

Metal oxides as a biostimulator for upregulation of genes involved in the biosynthesis of Rebaudioside- A.

Stevia rebaudiana Bertoni is a kind of perennial medicinal plant with sweetening properties which belongs to Asteraceae family. Its leaves with fundamental glycoside compounds consist of both a sugar part and a non-sugar sector. One of the glycoside compounds is Rebaudioside- A which has a greater importance in business. This experiment was conducted to evaluate the effects of Ag2O, CrO3, PbO, Fe2O3, BaO and TiO2 on the expression pattern of these genes in the Stevia rebaudiana. Rebaudioside- A biosynthesis was repeated 3 times with concentrations of 50, 100 and 200µM. Also, the results of the study pertaining to the expression pattern of these genes showed that metal oxides have led to an increase in the expression of the regulatory genes involved in biosynthesis of Rebaudioside- A. According to the expression profile, it was found that its effect on DXR, HDS, HDR, IDI and CPPS genes is more than other genes. The peak HPLC indicated for stevioside and Rebaudioside- A represents an increase in the production of this active ingredient under the influence of all treatments. In general, the expression profile of these genes and the results of HPLC show that whatever going to the end of the pathway of production of Rebaudioside- A, the activity of the enzymes increases under the influence of these treatments, and eventually a greater amount of Rebaudioside- A will be produced. This process shows that metal oxides will have a significant effect on the biosynthesis of Rebaudioside- A.

Monitoring ZEO apoptotic potential in 2D and 3D cell cultures and associated spectroscopic evidence on mode of interaction with DNA.

Recognizing new anticancer compounds to improve Breast cancer treatment seems crucial. Essential oil of Zataria Multiflora (ZEO) is a secondary metabolite with some biological properties, yet underlying cellular and molecular anticancer properties of ZEO is unclear. GC/MS analysis revealed that carvacrol is the major ingredient of the essential oil. ZEO increasingly suppressed viability in MDA-MB-231, MCF-7 and T47D Breast cancer cells while nontoxic to L929 normal cells in monolayer cell cultures (2D), whereas MDA-MB-231 multicellular spheroids (3D) were more resistant to inhibition. ZEO significantly induced cell apoptosis confirmed by fluorescent staining, flow cytometry analysis and DNA fragmentation in MDA-MB-231 2D and 3D cell cultures. ZEO increased ROS generation and subsequent loss of ΔΨm, caspase 3 activation and DNA damage which consequently caused G1 and G2/M cell cycle arrest in a dose- and time-dependent manner in 2D. S phase arrest occurred in cell spheroids therefore ZEO possible DNA interaction with gDNA was investigated and revealed ZEO binds DNA via intercalation. Altogether, these data corroborate anticancer properties of ZEO and suggest that cell culture format (2D monolayer vs. 3D spheroid) plays a critical role in drug response and provides new insights into the mechanisms underlying ZEO cytotoxicity effect on Breast cancer cells.

Preparation and characterization of potato starch-thymol dispersion and film as potential antioxidant and antibacterial materials.

The antioxidant/antimicrobial capacity and physical properties of potato starch dispersions enriched with polysorbate-thymol micelle were investigated. Results showed that potato starch have radical scavenging and antibacterial activities only in the presence of polysorbate-thymol but with lower level than polysorbate-thymol alone. The decrease in the antioxidant and antibacterial activities may be attributed to the encapsulation of thymol in the starch chain. Polysorbate-thymol caused a decrease in the particle size and viscosity and an increase in the zeta potential of the starch dispersions. Futhermore, polysorbate-thymol leads to a decrease in the tensile strength, rigidity and swelling, and an increase in the flexibility, solubility and water vapour permeability of the starch films. Atomic force microscopy revealed polysorbate-thymol micelles in the film matrix enclosed by polysaccharide crystal. Scanning electron microscope analysis indicated that polysorbate-thymol caused a coarse film microstructure with the distributed crack in the film matrix. Based on the results, the antioxidant and antibacterial activities of the starch-polysorbate-thymol make starch film suitable for food packaging and preservation.

Chemical composition, antioxidant and antimicrobial activities of essential oil obtained from Ferula assa-foetida oleo-gum-resin: effect of collection time.

The properties of essential oils obtained from Ferula assa-foetida oleo-gum-resins (OGRs) collectioned in three collections times in 15 June (OGR1), 30 June (OGR2) and 15 July (OGR3) 2011 was investigated. Essential oil from OGR1 was constituted high levels of (E)-1-propenyl sec-butyl disulfide (23.9%) and 10-epi-γ-eudesmol (15.1%). Essential oil from OGR2 was constituted high levels of (Z)-1-propenyl sec-butyl disulfide (27.7%) and (E)-1-propenyl sec-butyl disulfide (20.3%). Essential oil from OGR3 was constituted high levels of ß-pinene (47.1%) and α-pinene (21.3%). Inhibitory concentration (IC50) for radical scavenging were 0.012-0.035, 0.025-0.047 and 0.035-0.066 mg/ml of essential oil obtained from OGR1, OGR2 and OGR3, respectively. Minimal inhibitory concentration (MIC) for Gram-positive and Gram-negative bacteria and fungi grpwth were 0.028-0.111, 0.027-0.107 and 0.018-0.058 mg/ml of essential oil obtained from OGR1, OGR2 and OGR3, respectively. Essential oils obtained from different OGRs have different composition and biological activity thus have different applications in food and health industry.

Evaluation of antioxidant and antimicrobial activities of essential oils from Carum copticum seed and Ferula assafoetida latex.

UNLABELLED: Carum copticum and Ferula assafoetida have several medicinal properties including antispasmodic, carminative, sedative, analgesic, and antiseptic. Reactive oxygen species (ROS), reactive nitrogen species (RNS), hydrogen peroxide (H(2) O(2) ), and thiobarbituric acid reactive substances (TBARS) scavenging activities of Carum and Ferula oils along with their antibacterial and antifungal activities were examined. Thymol (40.25%), γ-terpinene (38.7%) and p-cymene (15.8%) were detected as the main components of Carum oil while, ß-pinene (47.1%), α-pinene (21.36%), and 1, 2-dithiolane (18.6%) were the main components of Ferula oil. Inhibitory concentrations (IC50) for total radical scavenging were between 40 and 60 and 130 and 160 µg/mL of Carum and Ferula oil, respectively. Minimal inhibitory concentration (MIC) for Salmonella typhi, Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Aspergillus niger, and Candida albicans were 78 ± 8, 65 ± 7, 14 ± 3, 5 ± 2, 5.6 ± 1.3, and 8.8 ± 2.2 µg/mL of Carum oil, respectively. MIC for S. typhi, E. coli, S. aureus, B. subtilis, A. niger, and C. albicans were >200, >200, 125 ± 17, 80 ± 12, 85 ± 5, and 90 ± 11 µg/mL of Ferula oil, respectively. Accordingly, Carum and Ferula oils could be used as safe and effective natural antioxidants to improve the oxidative stability of fatty foods during storage and to preserve foods against food burn pathogens. PRACTICAL APPLICATION: This study clearly demonstrates the potential of Carum and Ferula oil especially Carum oil as natural antioxidant and antimicrobial agent. The chemical composition of essential oils was identified. Thus, identification of such compounds also helps to discover of new antioxidant, antibacterial and antifungal agents for potential applications in food safety and food preservation.

Anti-inflammatory effect of Mentha longifolia in lipopolysaccharide-stimulated macrophages: reduction of nitric oxide production through inhibition of inducible nitric oxide synthase.

Mentha longifolia is an aromatic plant used in flavoring and preserving foods and as an anti-inflammatory folk medicine remedy. The present study assessed the effects of M. longifolia extracts, including essential oil and crude methanol extract and its fractions (ethyl acetate, butanol and hexane), on nitric oxide (NO) production and inducible NO synthase (iNOS) mRNA expression in lipopolysaccharide (LPS)-stimulated J774A.1 cells using real-time polymerase chain reaction (PCR). The cytotoxic effects of the extracts on the cells were examined and non-cytotoxic concentrations (<0.2 mg/ml) were used to examine their effects on NO production and iNOS mRNA expression. Only the hexane fraction that contained high levels of phenolic and flavonoid compounds at concentrations from 0.05-0.20 mg/ml significantly reduced NO production in LPS-stimulated cells (p < 0.001). Real-time PCR analysis indicated the ability of this fraction at the same concentrations to significantly decrease iNOS as well as TNFα mRNA expression in the cells (p < 0.001). All extracts were able to scavenge NO radicals in a concentration-dependent manner. At concentrations greater than 0.2 mg/ml, total radicals were 100% scavenged. In conclusion, M. longifolia possibly reduces NO secretion in macrophages by scavenging NO and inhibiting iNOS mRNA expression, and also decreases TNFα pro-inflammatory cytokine expression, thus showing its usefulness in the inflammatory disease process.

Inhibitory effects of Zataria multiflora essential oil and its main components on nitric oxide and hydrogen peroxide production in lipopolysaccharide-stimulated macrophages.

OBJECTIVES: Zataria multiflora is an aromatic plant that is used in flavouring and preserving foods and also used as an antispasmodic, anaesthetic and antinociceptive agent. In this study, the effects of Z. multiflora essential oil on nitric oxide (NO) and hydrogen peroxide (H(2) O(2) ) production in lipopolysaccharide (LPS)-stimulated macrophages was investigated. METHODS: Z. multiflora essential oil was extracted by water-distillation, analysed by GC-MS and then the effect of the essential oil on NO and H(2) O(2) production was investigated. KEY FINDINGS: Carvacrol (52%), thymol (16%) and p-cymene (10%) were the main components of the oil. The IC50 (concentration providing 50% inhibition) for reactive oxygen scavenging was estimated to be 5.7, 3 and 4.2 µg/ml for the essential oil, thymol and carvacrol, respectively, while the corresponding IC50 values for reactive nitrogen scavenging were estimated to be 8.6, 4.7 and 6.6 µg/ml. Z. multiflora essential oil, thymol, and carvacrol significantly reduced NO and H(2) O(2) production as well as NO synthase and NADH oxidase activity in LPS-stimulated murine macrophages while p-cymene did not show any antioxidant activity. CONCLUSIONS: Z. multiflora essential oil has the potential to be used in the therapy of oxidative damage.