ABSTRACT
We evaluated whether thymol (THY) (30â¯mg/kg b.wt) could relieve the adverse effects of the neonicotinoid insecticide imidacloprid (IMD) (22.5â¯mg/kg b.wt) on the liver in a 56-day oral experiment and the probable underlying mechanisms. THY significantly suppressed the IMD-associated increase in hepatic enzyme leakage. Besides, the IMD-induced dyslipidemia was considerably corrected by THY. Moreover, THY significantly repressed the IMD-induced hepatic oxidative stress, lipid peroxidation, DNA damage, and inflammation. Of note, the Feulgen, mercuric bromophenol blue, and PAS-stained hepatic tissue sections analysis declared that treatment with THY largely rescued the IMD-induced depletion of the DNA, total proteins, and polysaccharides. Moreover, THY treatment did not affect the NF-kB p65 immunoexpression but markedly downregulated the Caspase-3 in the hepatocytes of the THY+IMD-treated group than the IMD-treated group. Conclusively, THY could efficiently protect against IMD-induced hepatotoxicity, probably through protecting cellular macromolecules and antioxidant, antiapoptotic, and anti-inflammatory activities.
Subject(s)
Caspase 3 , DNA Damage , Insecticides , Liver , NF-kappa B , Neonicotinoids , Nitro Compounds , Oxidative Stress , Signal Transduction , Thymol , Animals , Neonicotinoids/toxicity , Oxidative Stress/drug effects , Liver/drug effects , Liver/metabolism , Liver/pathology , DNA Damage/drug effects , Nitro Compounds/toxicity , Caspase 3/metabolism , Caspase 3/genetics , Male , Thymol/pharmacology , Thymol/toxicity , Insecticides/toxicity , Signal Transduction/drug effects , NF-kappa B/metabolism , Rats, Wistar , Rats , Lipid Peroxidation/drug effectsABSTRACT
The excessive use of herbicides has caused a series of problems related to human health, environmental pollution, and an increase in the resistance of plants to commercial herbicides. As an alternative, natural compounds and their semisynthetic derivatives have been widely studied to obtain environmentally friendly and more effective herbicides than the usual ones. In view of these factors, the aim of this work was to synthesize new molecules with herbicidal potential using thymol as a starting material, a natural phenol that has a pronounced phytotoxic effect. Novel N-phenyl-2-thymoxyacetamides were synthesized and characterized by MS and by 1H and 13C NMR. All prepared molecules were subjected to phytotoxic and cytotoxic activity assays using Lactuca sativa L. and Sorghum bicolor L. as model plants. Molecules containing chlorine in the para position of the thymoxy group exhibited phytotoxic and cytogenotoxic effects superior to those of the commercial herbicides 2,4-D and glyphosate.
Subject(s)
Herbicides , Thymol , Humans , Thymol/toxicity , Herbicides/toxicity , Glyphosate , PlantsABSTRACT
The sustainability of the traditional extensive livestock sector will only be possible if healthy dung-decomposing insect communities are preserved. However, many current pharmaceutical anthelmintics are harmful to dung beetles, their presence can have a negative impact on biological systems. Phytochemical anthelmintics are an alternative to ecotoxic synthetic pharmaceutical anthelmintics, although ecotoxicological tests of their possible indirect effects on dung beetles are required to demonstrate their viability. In this study, the potential ecotoxicity of thymol, carvacrol, cinnamaldehyde and garlic oil (diallyl disulfide and diallyl trisulfide) were tested for the first time. Inhibition of antennal response was measured as a relevant parameter by obtaining relevant toxicity thresholds derived from concentrationâresponse curves, such as the IC50. All phytochemical compounds tested were demonstrated to be suitable alternative candidates to the highly ecotoxic compound ivermectin, considering their non-toxicity to nontarget organisms. Residues of the phytochemical antiparasitics found in cattle droppings were extremely low, even undetectable in the case of diallyl disulfide and diallyl trisulfide. Furthermore, our results showed that none of the phytochemical compounds have ecotoxic effects, even at extremely high concentrations, including those almost 1000 times higher than what is most likely to be found in dung susceptible to ingestion by dung beetles in the field. We can conclude that the four selected phytochemical compounds meet the requirements to be considered reliable alternatives to ecotoxic veterinary medicinal products, such as ivermectin.
Subject(s)
Anthelmintics , Coleoptera , Animals , Cattle , Anthelmintics/toxicity , Coleoptera/drug effects , Ivermectin/toxicity , Phytochemicals/toxicity , Thymol/toxicityABSTRACT
Zinc oxide nanoparticles (ZnO-NPs) have many exciting properties that make their use in a continuous increase in various biomedical, industrial, and agricultural applications. This is associated with accumulation in the aquatic ecosystems and fish exposure with consequent deleterious effects. To determine the potential of thymol to counteract the immunotoxic effects of ZnO-NPs, Oreochromis niloticus was exposed to ZnO-NPs (â LC50 =1.14 mg/L, for 28 days) with or without feeding a thymol-incorporated diet (1 or 2 g/kg diet). Our data demonstrated a reduction of aquaria water quality, leukopenia, and lymphopenia with a decrease in serum total protein, albumin, and globulin levels in exposed fish. At the same time, the stress indices (cortisol and glucose) were elevated in response to ZnO-NPs exposure. The exposed fish also revealed a decline in serum immunoglobulins, nitric oxide, and the activities of lysozyme and myeloperoxidase, in addition to reduced resistance to the Aeromonas hydrophila challenge. The RT-PCR analysis showed downregulation of antioxidant (SOD) superoxide dismutase and (CAT) catalase gene expression in the liver tissue with overexpression of the immune-related genes (TNF-α and IL-1ß). Importantly, we found that thymol markedly protected against ZnO-NPs-induced immunotoxicity in fish co-supplemented with thymol (1 or 2 g/kg diet) in a dose-dependent manner. Our data confirm the immunoprotective and antibacterial effects of thymol in ZnO-NPs exposed fish, supporting the potential utility of thymol as a possible immunostimulant agent.
Subject(s)
Cichlids , Fish Diseases , Nanoparticles , Water Pollutants, Chemical , Zinc Oxide , Animals , Cichlids/metabolism , Aeromonas hydrophila , Zinc Oxide/toxicity , Zinc Oxide/metabolism , Thymol/toxicity , Thymol/analysis , Thymol/metabolism , Ecosystem , Water Pollutants, Chemical/toxicity , Dietary Supplements/analysis , Diet/veterinary , Antioxidants/metabolism , Disease Resistance , Animal Feed/analysisABSTRACT
Extensive usage of synthetic pesticides has proved to be destructive to all living being and the resurgence of pest resistance. Compounds derived from certain plants are usually safer compared to chemical control of pest. The present study thus intended to use Thymus vulgaris essential oil (EO) and two of its derivatives including thymol and carvacrol in order to see their deleterious effects on Glyphodes pyloalis (Walker). We also studied the oil components. This pest has recently become a serious concern for the silk industry. Our results showed that the thyme EO contain several components including thymol (26.9%), ρ-Cymene (14.54%), linalool (13.39%) and carvacrol (5.7%). Our toxicity tests revealed an estimated LD50 values for thyme EO, thymol and carvacrol 2.82, 32.18 and 56.54 µg/larva, respectively. However, the thyme EO was more toxic than its two tested compounds. The activity of certain detoxifying enzymes such as α- and ß-esterase, glutathione S-transferase and cytochrome P450 were significantly inhibited by thymol-treated larvae compared to the control group. Similarly, the activity of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase and alkaline phosphatases enzymes in thymol-treated larvae decreased while the activity of acid phosphatases increased. Our results suggest that thyme EO and its components have potential for the control of G. pyloalis larvae in mulberry orchards, where no synthetic chemicals are allowed.
Subject(s)
Morus , Oils, Volatile , Thymus Plant , Animals , Oils, Volatile/toxicity , Thymol/toxicity , Larva , Alkaline PhosphataseABSTRACT
Understanding the cumulative risk of chemical mixtures at environmentally realistic concentrations is a key challenge in honey bee ecotoxicology. Ecotoxicogenomics, including transcriptomics, measures responses in individual organisms at the molecular level which can provide insights into the mechanisms underlying phenotypic responses induced by one or more stressors and link impacts on individuals to populations. Here, fifth instar honey bee larvae were sampled from a previously reported field experiment exploring the phenotypic impacts of environmentally realistic chronic exposures of the pesticide imidacloprid (5 µg.kg-1 for six weeks) and the acaricide thymol (250 g.kg-1 applied via Apiguard gel in-hive for four weeks), both separately and in combination. RNA-seq was used to discover individual and interactive chemical effects on larval gene expression and to uncover molecular mechanisms linked to reported adult and colony phenotypes. The separate and combined treatments had distinct gene expression profiles which represented differentially affected signaling and metabolic pathways. The molecular signature of the mixture was characterised by additive interactions in canonical stress responses associated with oxidative stress and detoxification, and non-additive interactions in secondary responses including developmental, neurological, and immune pathways. Novel emergent impacts on eye development genes correlated with long-term defects in visual learning performance as adults. This is consistent with these chemicals working through independent modes of action that combine to impact common downstream pathways, and highlights the importance of establishing mechanistic links between molecular and phenotypic responses when predicting effects of chemical mixtures on ecologically relevant population outcomes.
Subject(s)
Insecticides , Thymol , Animals , Bees/genetics , Insecticides/toxicity , Larva , Neonicotinoids/toxicity , Nitro Compounds , Phenotype , Thymol/toxicityABSTRACT
The cytotoxicity of carvacrol- and thymol-functionalised mesoporous silica microparticles (MCM-41) was assessed in the human hepatocarcinoma cell line (HepG2). Cell viability, lactate dehydrogenase (LDH) activity, reactive oxygen species (ROS) production, mitochondrial membrane potential (ΔΨm), lipid peroxidation (LPO) and apoptosis/necrosis analyses were used as endpoints. The results showed that both materials induced cytotoxicity in a time- and concentration-dependent manner, and were more cytotoxic than free essential oil components and bare MCM-41. This effect was caused by cell-particle interactions and not by degradation products released to the culture media, as demonstrated in the extract dilution assays. LDH release was a less sensitive endpoint than the MTT (thiazolyl blue tetrazolium bromide) assay, which suggests the impairment of the mitochondrial function as the primary cytotoxic mechanism. In vitro tests on specialised cell functions showed that exposure to sublethal concentrations of these materials did not induce ROS formation during 2 h of exposure, but produced LPO and ΔΨm alterations in a concentration-dependent manner when cells were exposed for 24 h. The obtained results generally support the hypothesis that the carvacrol- and thymol-functionalised MCM-41 microparticles induced toxicity in HepG2 cells by an oxidative stress-related mechanism that resulted in apoptosis through the mitochondrial pathway.
Subject(s)
Cymenes/toxicity , Silicon Dioxide/toxicity , Thymol/toxicity , Apoptosis/drug effects , Cell Survival/drug effects , Cymenes/chemistry , Hep G2 Cells , Humans , Lipid Peroxidation/drug effects , Membrane Potential, Mitochondrial/drug effects , Mitochondria/drug effects , Mitochondria/metabolism , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Silicon Dioxide/chemistry , Thymol/chemistry , ToxicologyABSTRACT
The extensive use of essential oil components in an increasing number of applications can substantially enhance exposure to these compounds, which leads to potential health and environmental hazards. This work aimed to evaluate the toxicity of four widely used essential oil components (carvacrol, eugenol, thymol, vanillin) using the in vivo model Caenorhabditis elegans. For this purpose, the LC50 value of acute exposure to these components was first established; then the effect of sublethal concentrations on nematodes' locomotion behaviour, reproduction, heat and oxidative stress resistance and chemotaxis was evaluated. The results showed that all the components had a concentration-dependent effect on nematode survival at moderate to high concentrations. Carvacrol and thymol were the two most toxic compounds, while vanillin had the mildest toxicological effect. Reproduction resulted in a more sensitive endpoint than lethality to evaluate toxicity. Only pre-exposure to carvacrol and eugenol at the highest tested sublethal concentrations conferred worms oxidative stress resistance. However, at these and lower concentrations, both components induced reproductive toxicity. Our results evidence that these compounds can be toxic at lower doses than those required for their biological action. These findings highlight the need for a specific toxicological assessment of every EOC application.
Subject(s)
Caenorhabditis elegans/drug effects , Eugenol/toxicity , Oils, Volatile , Thymol/toxicity , Animals , Benzaldehydes/toxicity , Cymenes/toxicity , Larva/drug effects , Lethal Dose 50 , Oils, Volatile/chemistry , Oils, Volatile/toxicityABSTRACT
Phenolic compounds carvacrol, thymol, eugenol, and vanillin are four of the most thoroughly investigated essential oil components given their relevant biological properties. These compounds are generally considered safe for consumption and have been used in a wide range of food and non-food applications. Significant biological properties, including antimicrobial, antioxidant, analgesic, anti-inflammatory, anti-mutagenic, or anti-carcinogenic activity, have been described for these components. They are versatile molecules with wide-ranging potential applications whose use may substantially increase in forthcoming years. However, some in vitro and in vivo studies, and several case reports, have indicated that carvacrol, thymol, and eugenol may have potential toxicological effects. Oxidative stress has been described as the main mechanism underlying their cytotoxic behavior, and mutagenic and genotoxic effects have been occasionally observed. In vivo studies show adverse effects after acute and prolonged carvacrol and thymol exposure in mice, rats, and rabbits, and eugenol has caused pulmonary and renal damage in exposed frogs. In humans, exposure to these three compounds may cause different adverse reactions, including skin irritation, inflammation, ulcer formation, dermatitis, or slow healing. Toxicological vanillin effects have been less reported, although reduced cell viability after exposure to high concentrations has been described. In this context, the possible risks deriving from increased exposure to these components for human health and the environment should be thoroughly revised.
Subject(s)
Oils, Volatile , Animals , Anti-Bacterial Agents , Eugenol/adverse effects , Eugenol/toxicity , Mice , Monoterpenes/adverse effects , Monoterpenes/toxicity , Oils, Volatile/adverse effects , Oils, Volatile/toxicity , Rabbits , Rats , Thymol/adverse effects , Thymol/toxicityABSTRACT
Among natural products, essential oils from aromatic plants have been reported to possess potent anticancer properties. In this work, we aimed to perform the cytotoxic concentration range screening and antiproliferative activity screening of chemically characterized Thymus vulgaris L. essential oil. In vivo bioassay was conducted using the brine shrimp lethality test (BSLT). In vitro evaluation of antiproliferative activity was carried out on three human tumor cell lines: breast adenocarcinoma MCF-7, lung carcinoma H460 and acute lymphoblastic leukemia MOLT-4 using MTT assay. Essential oil components thymol (36.7%), p-cymene (30.0%), γ-terpinene (9.0%) and carvacrol (3.6%) were identified by gas chromatography/mass spectrometry. Analyzed essential oil should be considered as toxic/highly toxic with LC50 60.38 µg/mL in BSLT and moderate/weakly cytotoxic with IC50 range 52.65-228.78 µg/mL in vitro, according to evaluated cytotoxic criteria. Essential oil induced a dose-dependent inhibition of cell proliferation in all tested tumor cell lines and showed different sensitivity. Dose dependent toxicity observed in bioassay as well as the in vitro assay confirmed that brine shrimp lethality test is an adequate method for preliminary toxicity testing of Thymus vulgaris L. essential oil in tumor cell lines.
Subject(s)
Antineoplastic Agents, Phytogenic/toxicity , Artemia/drug effects , Oils, Volatile/toxicity , Plant Oils/toxicity , Thymus Plant/chemistry , Animals , Antineoplastic Agents, Phytogenic/pharmacology , Artemia/growth & development , Biological Assay , Cell Division/drug effects , Cell Line, Tumor/drug effects , Cymenes/pharmacology , Cymenes/toxicity , Dose-Response Relationship, Drug , Drug Screening Assays, Antitumor , Gas Chromatography-Mass Spectrometry , Humans , Inhibitory Concentration 50 , Larva/drug effects , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Plant Oils/chemistry , Plant Oils/pharmacology , Thymol/pharmacology , Thymol/toxicityABSTRACT
Thymol and carvacrol are phenolic isomers with the potential developmental toxicity and endocrine disruptions (ED) at low concentrations. However, few reports estimated their toxicity and ED below 10-6 M (150 µg/L) (MW of thymol and carvacrol: 150 g/mol). In this study, both chemicals were determined for the developmental toxicity and potential ED at 500 µg/kg and 50 µg/kg using the chicken embryonic assay, potential estrogenic activity (EA) at 10-12 to 10-7 M (1.5 × 10-4 to 15 µg/L) by the MCF-7 cell proliferation assay, mutagenicity at 10-12 to 10-6 M (1.5 × 10-4 to 150 µg/L) by the Ames test, and an in silico method for ED. Carvacrol showed mutagenic risks at 10-7, 10-8, and 10-11 M (15, 1.5, and 0.0015 µg/L) while thymol at 10-6 and 10-8 M (150 and 1.5 µg/L). Carvacrol negatively impacted embryonic growth at 50 µg/kg, with weak EA at 10-8 M (1.5 µg/L). Carvacrol but not thymol had weak EA at 10-12 M (1.5 × 10-4 µg/L). Molecular docking to 14 types of hormone-related receptors revealed that carvacrol had higher binding affinities to two estrogen receptors and the mineralocorticoid receptor than those to thymol. Carvacrol and thymol varied in toxicities due to a different location of one phenolic hydroxyl group.
Subject(s)
Cymenes/toxicity , Estrogens/toxicity , Thymol/toxicity , Animals , Chick Embryo , Cymenes/administration & dosage , Cymenes/chemistry , Estradiol/chemistry , Estradiol/pharmacology , Humans , MCF-7 Cells , Molecular Docking Simulation , Molecular Structure , Mutagenesis , Protein Binding , Receptors, Estrogen , Thymol/administration & dosage , Thymol/chemistryABSTRACT
Thymol, a monoterpene phenol, is used as a natural biocide. To circumvent its chemical instability, we propose use of thymol-loaded biogenic silica nanoparticles (BSiO2 #THY NPs); however, the toxicity of this system for aquatic organisms is unknown. Thus, the present study aimed to evaluate the toxicogenetic effects induced by thymol, BSiO2 NP, and BSiO2 #THY on Artemia salina and zebrafish (Danio rerio) early life stages. We also investigated the impact of BSiO2 aggregation in different exposure media (saline and freshwater). Based on the median lethal concentration at 48 h (LC5048h ), BSiO2 #THY (LC5048h = 1.06 mg/L) presented similar toxic potential as thymol (LC5048h = 1.03 mg/L) for A. salina, showing that BSiO2 had no influence on BSiO2 #THY toxicity. Because BSiO2 aggregated and sedimented faster in A. salina aqueous medium than in the other medium, this NP had lower interaction with this microcrustacean. Thus, BSiO2 #THY toxicity for A. salina is probably due to the intrinsic toxicity of thymol. For zebrafish early life stages, BSiO2 #THY (LC5096h = 13.13 mg/L) was more toxic than free thymol (LC5096h = 25.60 mg/L); however, BSiO2 NP has no toxicity for zebrafish early life stages. The lower aggregation of BSiO2 in the freshwater medium compared to the saline medium may have enhanced thymol's availability for this aquatic organism. Also, BSiO2 #THY significantly induced sublethal effects as thymol, and both were genotoxic for zebrafish. In conclusion, although BSiO2 #THY still needs improvements to ensure its safety for freshwater ecosystems, BSiO2 NP seems to be a safe nanocarrier for agriculture. Environ Toxicol Chem 2021;40:333-341. © 2020 SETAC.
Subject(s)
Nanoparticles , Water Pollutants, Chemical , Animals , Ecosystem , Nanoparticles/toxicity , Silicon Dioxide/toxicity , Thymol/toxicity , Water Pollutants, Chemical/toxicity , ZebrafishABSTRACT
In the present study, ternary blends based on poly (lactic acid)/poly (ε-caprolactone)/thermoplastic starch were prepared at different concentrations of synthesized zinc oxide nanoparticles (ZnO-NPs) and thymol. The sizes of ZnO-NPs with an average diameter of about 30-50 nm were detected by FE-SEM analysis. Moreover, the effect of ZnO-NPs and thymol on morphological, FT-IR spectrum, UV absorption, thermal stability, cytotoxicity, and antibacterial properties of neat blend was investigated. TGA analysis showed that the addition of ZnO-NPs and/or thymol diminished thermal stability of the system. Incorporating ZnO-NPs improved antibacterial activities of the neat blend, but MTT-assay and AO fluorescent staining test results depicted a decrease in cell viability to less than 20% by the addition of 5 wt% ZnO-NPs. In such a condition, the addition of thymol to the nanocomposites exhibited a dose-dependent increase in cell survival mostly due to thymol antioxidant properties. Interestingly, the antibacterial performance of compounds was also improved by the presence of thymol. Therefore, the obtained nanocomposites have potential to extend applications of innovative biomedical devices for future research in which both high cell viability and superior antibacterial properties are needed such as an antibacterial wound healing film.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antioxidants/pharmacology , Biocompatible Materials/pharmacology , Nanocomposites , Polyesters/pharmacology , Thymol/pharmacology , Zinc Oxide/pharmacology , Animals , Anti-Bacterial Agents/toxicity , Antioxidants/toxicity , Biocompatible Materials/toxicity , Drug Evaluation, Preclinical , Escherichia coli/drug effects , Fibroblasts/drug effects , Materials Testing , Mice , Microscopy, Electron, Scanning , Nanocomposites/toxicity , Polyesters/toxicity , Spectrophotometry, Ultraviolet , Spectroscopy, Fourier Transform Infrared , Staphylococcus aureus/drug effects , Thermogravimetry , Thymol/toxicity , X-Ray Diffraction , Zinc Oxide/toxicityABSTRACT
Thymol is a natural phenolic monoterpene widely produced by different species belonging to the Labiateae family. Although the thymol phytotoxicity is well known, the knowledge of its potential toxic mechanism is still limited. In this regard, the model species Arabidopsis thaliana was treated for 16 days by sub-irrigation with 300 µM of thymol. The results confirmed the high phytotoxic potential of this phenolic compound, which caused a reduction in plant growth and development. Thymol induced a water status alteration accompanied by an increase in ABA content and stomatal closure. Furthermore, leaves appeared necrotic in the margins and their temperature rinsed. The increase in H2O2 content suggested an oxidative stress experienced by treated plants. Both metabolomic and proteomic analysis confirmed this hypothesis showing a strong increase in osmoprotectants content, such as galactinol and proline, and a significant up-accumulation of proteins involved in ROS detoxification. Furthermore, the down-accumulation of proteins and pigments involved in the photosynthetic machinery, the increase in light sensitivity and the lower PSII efficiency well indicated a reduction in photosynthetic activity. Overall, we can postulate that thymol-induced phytotoxicity could be related to a combined osmotic and oxidative stress that resulted in reduced plant development.
Subject(s)
Arabidopsis/drug effects , Metabolome , Proteome , Thymol/toxicity , Abscisic Acid , Arabidopsis/growth & development , Hydrogen Peroxide , Osmotic Pressure , Oxidative Stress , Photosynthesis , Plant LeavesABSTRACT
The effects of Ajwain essential oil and its major constituents were investigated against newly fourth instar larvae of Tuta absoluta, a major pest of tomato cropping in Iran and other parts of the world. The study included individual efficacy of each compound and determination of their synergistic or antagonistic efficacy. We also determined the activities of detoxifying enzymes in treated insects. According to the results, the Ajwain essential oil and thymol showed the highest toxicity against T. absoluta larvae (LC50 = 6.14 and 7.72 µL/mL, respectively, and LC30 = 3.05 and 3.57 µL/mL, respectively). The ɤ-terpinene and p-cymene had lesser toxicity compared to other compounds. The larvicidal potency increased by binary mixtures of active constituents, such as thymol + p-cymene and thymol + ɤ-terpinene. On the other hand, ɤ-terpinene + p-cymene mixture demonstrated antagonistic effect on the larvae. The esterases and glutathione S-transferases were significantly increased in all treatments. Finally, our results revealed a significant inhibition of AChE activity in the treated larvae by all treatments except for ɤ-terpinene + p-cymene. In vitro experiments representing AChE inhibition with IC50 values were recorded 0.370, 0.457, 0.528, 1.094 and 1.323 µL/mL for thymol + p-cymene, thymol + ɤ-terpinene, thymol, ɤ-terpinene and p-cymene, respectively. Thymol demonstrated significant potential as a controlling agent of T. absoluta larvae, both larvicidal, compatible with other compounds with strong AChE inhibition properties. These finding could pave the way for development of new botanicals based on EO constituents which should be completed with preparation of an efficient formulation and field trials.
Subject(s)
Apiaceae , Lepidoptera/drug effects , Oils, Volatile/toxicity , Animals , Carum , Cymenes , Iran , Larva/drug effects , Solanum lycopersicum , Moths/drug effects , Thymol/toxicityABSTRACT
Epinephrine HFA (Primatene® Mist) is a newly formulated asthma metered dose inhaler developed to replace the previous Primatene® Mist CFC. The formulation of Epinephrine HFA contains thymol, a substance recognized to be safe by the FDA. Although the content of thymol contained in Epinephrine HFA is much lower compared to many common foods and medications available, there are no known nonclinical data about the chronic toxicity of thymol through inhalation. Two sequential 6-month studies of identical design were conducted to assess the chronic toxicity of inhaled thymol in mice. Four treatment groups, (a) Air; (b) vehicle control; (c) Article-1 (thymol 0.1%); and (d) Article-2 (thymol 0.5%) were assessed in 128 mice for 26 weeks. The mice were sacrificed at the end of the treatment period and a histopathologic evaluation was performed with respect to lungs, bronchial lymph nodes, nasal passages/nasopharynx, and trachea. Forty-five pathologic assessment parameters (PAPs) were evaluated. In total, 5591 data points from 487 mouse organs were assessed. Chronic toxicity index was calculated for 16 PAPs that had multiple histopathologic abnormal observations. The t tests were conducted for these 16 PAPs (Articles-1 and 2 versus Air and vehicle control, respectively), and all P-values were greater than .05 indicating no significant differences between all treatment groups. An evaluation was also conducted for 25 PAPs that had only a very small number of pathologic abnormalities. No significant differences for chronic toxicity were found when comparing mice under long-term repeated exposure of high doses of inhaled thymol and mice that inhaled no thymol.
Subject(s)
Epinephrine/chemistry , Lung/drug effects , Respiratory System/drug effects , Thymol/toxicity , Administration, Inhalation , Animals , Case-Control Studies , Female , Lung/cytology , Male , Mice , Models, Animal , Respiratory System/cytology , Toxicity Tests, ChronicABSTRACT
Thymol is a phytochemical component present in many plants used as food additive in order to promote animal growth due to its several biological properties. However, possible side effects of thymol remain poorly known limited to few reports. In this sense, we evaluated the enzymes of the purinergic signaling such as, ectonucleoside triphosphate diphosphohydrolase (NTPDase), 5'-nucleotidase and adenosine deaminase (ADA), that play an important role on toxicity induced by excessive adenosine triphosphate (ATP) content in the extracellular environment. Thus, the aim of this study was to evaluate whether purinergic signaling could be considered a potential target of thymol-induced inflammation, and the toxicity in tissues and immune cells of mice after thymol administration. NTPDase activity (ATP as substrate) in serum, spleen, and splenic lymphocytes was lower after 30 days of oral treatment at doses of 10, 20, and 40 mg/kg of thymol, while ADA activity was stimulated at 20 and 40 mg/kg. No differences were observed between groups regarding NTPDase (ADP as substrate) and 5'-nucleotidase activities in all evaluated tissues. Based on these evidences, adenine nucleotide hydrolysis is modified in serum, spleen, and splenic lymphocytes of mice treated with thymol, contributing to inflammation and toxicity by a reduction on ATP hydrolyses and its possible accumulation in the extracellular medium and increased Ado desamination and its possible reduction in the extracellular environment, leading to a self-sustained pro-inflammatory deleterious cycle. In summary, all tested thymol concentrations induced inflammation and toxicity in tissues and immune cells of treated mice.
Subject(s)
Adenosine Deaminase/metabolism , Antigens, CD/metabolism , Apyrase/metabolism , Inflammation/pathology , Lymphocytes/drug effects , Purines/metabolism , Spleen/drug effects , Thymol/toxicity , Adenosine Triphosphate , Animals , Anti-Infective Agents/toxicity , Inflammation/chemically induced , Inflammation/metabolism , Lymphocytes/metabolism , Lymphocytes/pathology , Male , Mice , Spleen/metabolism , Spleen/pathologyABSTRACT
Several evidences have related the biochemical and pharmacological properties of thymol, but the possible neurotoxic effects of this compound remain unknown and not evaluated. Thus, the purpose of this study was to evaluate whether intake of thymol in different doses (10, 20 and 40â¯mg/kg) induce neurotoxicity and behavioral alterations using mice as experimental model, as well as the involvement of blood-brain barrier (BBB) and brain neurotransmitters in these alterations. Thymol (20 and 40â¯mg/kg) significantly decrease latency time to inhibitory avoidance task when compared to control group, indicating a memory loss after 30 days of oral treatment. Also, thymol (20 and 40â¯mg/kg) induced a significant increase on BBB permeability to Evan's blue dye when compared to control group, which is an indicative of BBB breakdown. Moreover, a significant increase of brain acetylcholinesterase (AChE) was observed in mice treated with 40â¯mg/kg of thymol, while the activity of sodium-potassium pump (Na+, K+-ATPase) was inhibited in mice treated with 20 and 40â¯mg/kg thymol when compared to control group. Finally, mice that received 20 and 40â¯mg/kg thymol showed a significant increase on cerebral reactive oxygen species (ROS) levels and cerebral xanthine oxidase (XO) activity compared to control group. Based on these evidences, the rupture of BBB can be considered an important pathway linked in thymol-induced memory loss. Also, the augmentation of brain ROS levels elicited by increase on XO activity may be a via involved in the damage to BBB, and an oxidative pathway that impairs the activity of brain neurotransmitters, as AChE and Na+, K+-ATPase. In summary, the dose of 10â¯mg/kg thymol can be safe and without neurotoxic effects in a period of 30 days of intake.
Subject(s)
Brain/drug effects , Memory Disorders/chemically induced , Neurotoxicity Syndromes , Thymol/toxicity , Acetylcholinesterase/metabolism , Administration, Oral , Animals , Behavior, Animal/drug effects , Brain/metabolism , Male , Memory Disorders/metabolism , Mice , Neurotoxicity Syndromes/metabolism , Permeability/drug effects , Reactive Oxygen Species/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism , Thymol/pharmacokinetics , Xanthine Oxidase/metabolismABSTRACT
Phenolic compounds of essential oils from the family Lamiaceae are commonly used substances in the food industry because of their flavouring, antimicrobial and antioxidant properties. In this context, it has become important to have healthy and safe products for consumers who are exposed to these phenolic compounds. The present study was aimed to investigate the toxic effects of carvacrol, thymol and their mixture on human gastric carcinoma (AGS) cells. Cells were analyzed after 24 h of exposure to different concentrations of carvacrol, thymol and their mixture by the ATP cell viability, 2',7' dichlorodihydrofluorescein diacetate (H2DCF-DA), reducte glutatione/oxide glutathione ((GSH)/GSSG-Glo) and comet assays. Apoptosis induction was studied by acridine orange/ethidium bromide staining and western blotting. Carvacrol, thymol and their mixture induced cytotoxicity, genotoxicity, apoptosis, increased reactive oxygen species (ROS) and decreased GSH levels after 24 h of their exposure in a dose-dependent manner. A close negative relationship was found between cell viability and ROS generation. We examined dose-dependent cytotoxic effects of carvacrol, thymol and their mixture in human AGS cells. Increased intracellular ROS causes oxidative stress in cells. The results indicated that these compounds should be used carefully in the food industry.