Herbal Melanin Inhibits Real-Time Cell Proliferation, Downregulates Anti-Apoptotic Proteins and Upregulates Pro-Apoptotic p53 Expression in MDA-MB-231 and HCT-116 Cancer Cell Lines.

Background and Objectives: Cancer is the second-most-important deadly disease in the world, leading to severe socioeconomic consequences and posing a public threat. Consequently, breast and colorectal cancers are significant cancer types that affect women and men more commonly, respectively. Treatment failure or recurrent diseases frequently occur due to resistance, in addition to the side effects of the currently available anticancer agents. Therefore, in this study, herbal melanin anticancer activity was investigated against human breast adenocarcinoma (MDA-MB-231) and human colorectal (HCT 116) cell proliferation and the expression of downregulated anti-apoptotic proteins and upregulated pro-apoptotic p53. Materials and Methods: MDA-MB-231 and HCT 116 cells were monitored for their real-time proliferation properties using Xcelligence. Herbal melanin of various concentrations significantly inhibited MDA-MB-231 and HCT 116 cell proliferation. Then, the expression of proapoptotic and anti-apoptotic proteins such as p53, Bcl-2 and Bcl-xl was studied using Western blotting. Results: The Bcl-2 and Bcl-xl expressions were downregulated, while the p53 expression was upregulated after treatment with herbal melanin. Similarly, the expression of apoptotic proteins such as Bcl-2, Bcl-xl, XIAP, Survivin, Bid, Bax, p53, Cytochrome C, PARP genes and mRNA was studied after herbal melanin treatment using real-time PCR, which revealed the downregulation of Bcl-2, Bcl-xl, XIAP and Survivin and the upregulation of Bid, Bax, p53, Cytochrome C and PARP apoptotic protein expression. Also, caspase 3 and 9 expressions were monitored after the treatment with herbal melanin, which revealed the upregulation of both the MDA-MB-231 and HCT 116 cell types. Conclusions: Overall, herbal melanin can be used as an alternative anticancer agent against the MDA-MB-231 and HCT 116 cell types.

Nimbin (N1) and analog N3 from the neem seeds suppress the migration of osteosarcoma MG-63 cells and arrest the cells in a quiescent state mediated via activation of the caspase-modulated apoptotic pathway.

BACKGROUND: Natural products are considered effective sources for new therapeutic research and development. The numerous therapeutic properties of natural substances in traditional medicine compel us to investigate the anti-cancer properties of Nimbin (N1) and its semi-natural analog Nimbic acid (N3) from Azadirachta indica against MG-63 Osteosarcoma cells. MATERIALS AND METHODS: The therapeutic efficacy of N1 and N3 were screened for their toxicity and cytotoxic activity using L6 myotubes, zebrafish larvae and MG-63 osteosarcoma cells. The mitochondrial membrane potential was evaluated using the Rhodamine 123 stain. Further, the nuclear and cellular damage was distinguished using Hoechst and Acridine orange/EtBr stain. The mechanism of cell cycle progression, cellular proliferation and caspase cascade activation was screened using scratch assay, flow cytometry, and mRNA expression analysis. RESULTS: The Nimbin and analogue N3 were found to be non-toxic to normal L6 cells (Rat skeletal muscles), exhibited cytotoxicity in MG-63 cells, and were exposed to be an active inhibitor of cell proliferation and migration. Analogs N1 and N3 induced negative mitochondrial membrane potential when stained with Rhodamine 123, leading to nuclear damage and apoptosis stimulation using AO/EtBr and Hoechst. Further, N1 and N3 induced cell cycle arrest in G0/G1 phase in flow cytometry using PI staining and induced apoptosis by activating the caspase cascade and upregulated Caspase 3 and caspase 9. CONCLUSION: The study demonstrated cytotoxic activity against MG-63 osteosarcoma cells while being non-toxic to normal L6 cells. These compounds inhibited cell proliferation and migration, induced mitochondrial dysfunction, nuclear damage, and apoptosis stimulation. Furthermore, N1 and N3 caused cell cycle arrest and activated the caspase cascade, ultimately leading to apoptosis. These findings indicate that N1 and N3 hold promise as potential candidates used alone or combined with existing drugs for further investigation and development as anti-cancer agents.

Hepatoprotective effect of dihydroxy piperlongumine in high cholesterol-induced non-alcoholic fatty liver disease zebrafish via antioxidant activity.

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are a growing epidemic and the most common liver diseases. Consumption of a western diet with high fats alters redox status, induces inflammation, and impairs the physiological function of hepatocytes. However, the pharmacological market lacks anti-NAFLD/NASH drugs. Long pepper (Piper longum L) is used in traditional Mongolian medicine for treating hyperlipidemia. Piperlongumine (PL) is a bioactive compound of Piper longum L, which usually possesses anticancer activities due to its ROS elevation property. However, when PL was demethylated they behave as an antioxidant. Previously, we found dihydroxy piperlongumine (DHPL) possesses high antioxidant activity among the hydroxy piperlongumines, which makes us curious to reveal the anti-NAFLD effect. A high-cholesterol diet (HCD) was chosen to induce NAFLD zebrafish model, and the antioxidant and lipid-lowering effects of DHPL were evaluated. Histological alterations of NAFLD were also scored along with gene expression to explore the molecular mechanism. DHPL reduced lipid accumulation in both short-term and long-term feeding trials. DHPL increases antioxidant activity and lipid-lowering gene expression and decreases hepatic triglyceride, oxidative stress, and lipogenic genes. In conclusion, DHPL halted the progression of HCD-induced NAFLD in the zebrafish model.

Antibacterial, Antioxidant, Larvicidal and Anticancer Activities of Silver Nanoparticles Synthesized Using Extracts from Fruits of Lagerstroemia speciose and Flowers of Couroupita guianensis.

The present study aimed to analyze the in vitro antibacterial, antioxidant, larvicidal and cytotoxicity properties of green synthesized silver nanoparticles (Ag NPs) using aqueous extracts from fruits of Lagerstroemia speciosa and flowers of Couropita guinensis. Synthesized Ag NPs were characterized using UV-DRS, FTIR, XRD, DLS, and High-Resolution SEM and TEM analyses. Absorption wavelength was observed at 386 nm by UV-DRS analysis and energy band gap was calculated as 3.24 eV. FTIR analysis showed the existence of various functional groups in the aqueous extract and in the NPs. DLS analysis showed the stability and particle size of the synthesized Ag NPs. SEM analysis revealed that Ag NPs are in a face centered cubic symmetry and spherical shape with a size of 23.9 nm. TEM analysis showed particle size as 29.90 nm. Ag NPs showed antibacterial activity against both Gram-positive and Gram-negative bacteria. DPPH scavenging trait of Ag NPs was ranging from 20.0 ± 0.2% to 62.4 ± 0.3% and observed significant larvicidal activity (LC50 at 0.742 ppm and LC90 at 6.061 ppm) against Culex quinquefasciatus. In vitro cytotoxicity activity of Ag NPs was also tested against human breast cancer (MCF-7) and fibroblast cells (L-929) and found that cells viabilities are ranging (500 to 25 µg/mL) from 52.5 ± 0.4 to 94.0 ± 0.7% and 53.6 ± 0.5 to 90.1 ± 0.8%, respectively. The synthesized Ag NPs have the potential to be used in the various biomedical applications.

Phyto-chemical and biological activity of Myristica fragrans, an ayurvedic medicinal plant in Southern India and its ingredient analysis.

The major aspects of using plant-derived medications are significantly safer and secure than synthetic ones. The n-hexane seed extract of ayurvedic medicinal plants Myristica fragrans was also utilized as food ingredients have analyzed for phytochemical existence by Gas Chromatography-mass spectrometry (GC-MS). Twenty-three phytoconstituents were identified with elemicin (24.44%) as the major constituent. Lipid peroxidase, catalase and DPPH assays were performed using the isolated elemicin and the results revealed significant antioxidant activity. The antibacterial study revealed that elemicin showed MIC of 31.25 µg/mL against Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi, and 62.5 µg/mL against Klebsiella pneumonia and Staphylococcus aureus. Elemicin exhibited better antifungal activity against Candida tropicalis and Aspergillus flavus than Aspergillus niger, Penicillium chrysogenum and Trichophyton rubrum. The study implies that the elemicin isolated from Myristica fragrans possess promising bioactive properties and can be crucially utilized in the development of therapeutic agents and food preservatives.

Inhibition of Aldose Reductase by Novel Phytocompounds: A Heuristic Approach to Treating Diabetic Retinopathy.

Aldose reductase (ALR2) activation in the polyol pathway has been implicated as the primary mechanism for the progression of diabetic retinopathy. Most of the aldose reductase inhibitors (ARIs), used for the treatment of diabetic complications, were withdrawn due to ineffective treatment and adverse side effects caused by nonspecificity. Epalrestat, a carboxylic acid inhibitor, is the only ARI used for the treatment of diabetic neuropathy, though associated with minor side effects to 8% of the treated population. Our study exploited the interactions of Epalrestat-ALR2 crystal structure for the identification of specific phytocompounds that could inhibit human lens ALR2. 3D structures of plant compounds possessing antidiabetic property were retrieved from PubChem database for inhibition analysis, against human lens ALR2. Among the shortlisted compounds, Agnuside and Eupalitin-3-O-galactoside inhibited lens ALR2 with IC50 values of 22.4 nM and 27.3 nM, respectively, compared to the drug Epalrestat (98 nM), indicating high potency of these compounds as ALR2 inhibitors. IC50 concentration of the identified ARIs was validated in vitro using ARPE-19 cells. The in silico and in vitro approaches employed to identify and validate specific and potent ALR2 inhibitors resulted in the identification of phytocompounds with potency equal to or better than the ALR2 inhibiting drug, Epalrestat.

In Vitro Screening of East Asian Plant Extracts for Potential Use in Reducing Ruminal Methane Production.

Indiscriminate use of antibiotics can result in antibiotic residues in animal products; thus, plant compounds may be better alternative sources for mitigating methane (CH4) production. An in vitro screening experiment was conducted to evaluate the potential application of 152 dry methanolic or ethanolic extracts from 137 plant species distributed in East Asian countries as anti-methanogenic additives in ruminant feed. The experimental material consisted of 200 mg total mixed ration, 20 mg plant extract, and 30 mL diluted ruminal fluid-buffer mixture in 60 mL serum bottles that were sealed with rubber stoppers and incubated at 39 °C for 24 h. Among the tested extracts, eight extracts decreased CH4 production by >20%, compared to the corresponding controls: stems of Vitex negundo var. incisa, stems of Amelanchier asiatica, fruit of Reynoutria sachalinensis, seeds of Tribulus terrestris, seeds of Pharbitis nil, leaves of Alnus japonica, stem and bark of Carpinus tschonoskii, and stems of Acer truncatum. A confirmation assay of the eight plant extracts at a dosage of 10 mg with four replications repeated on 3 different days revealed that the extracts decreased CH4 concentration in the total gas (7-15%) and total CH4 production (17-37%), compared to the control. This is the first report to identify the anti-methanogenic activities of eight potential plant extracts. All extracts decreased ammonia (NH3-N) concentrations. Negative effects on total gas and volatile fatty acid (VFA) production were also noted for all extracts that were rich in hydrolysable tannins and total saponins or fatty acids. The underlying modes of action differed among plants: extracts from P. nil, V. negundo var. incisa, A. asiatica, and R. sachalinensis resulted in a decrease in total methanogen or the protozoan population (p < 0.05) but extracts from other plants did not. Furthermore, extracts from P. nil decreased the population of total protozoa and increased the proportion of propionate among VFAs (p < 0.05). Identifying bioactive compounds in seeds of P. nil by gas chromatography-mass spectrometry analysis revealed enrichment of linoleic acid (18:2). Overall, seeds of P. nil could be a possible alternative to ionophores or oil seeds to mitigate ruminal CH4 production.

Silver nanoparticles containing stearic acid isolated from Catharanthus roseus: Ovicidal and oviposition-deterrent activities on Earias vittella and ecotoxicological studies.

In the recent past, many agrochemicals have been used to control pests, but many of these fail due to the development of resistance. Many researchers, therefore, concentrate on developing new pesticide formulations from natural resources (plants/microorganism). In the present study, different extracts from Catharanthus roseus (Madagascar periwinkle) was evaluated for their ovicidal and oviposition deterrent activities against Earias vittella (spiny bollworm). Among the tested extracts DCM (Dichloromethane) extract showed highest ovicidal activity (70.47%) and oviposition deterrent activity (75.41%) against E. vittella. Based on this biological activity, DCM extract was fractionated and isolated 7 fractions; all of these were evaluated for their ovicidal and oviposition deterrent activity against E. vittella. Maximum ovicidal and oviposition deterrent activity was recorded in fraction 5, followed by the 7th fraction. Stearic acid was isolated from fraction 5 and was subjected to nanoparticle synthesis. This nanoparticle was tested for its effects against E. vittella. It was found to exhibit 100% oviposition deterrent and 95% ovicidal activities against E. vittella, and also reduced the protein (53.63%), glutothionine esterase (39.16%), and esterase activity (45.25%) of the treated larvae. The synthesized nanoparticle was subjected to ecotoxicology evaluation against Daphnia sp. (water fleas) and Cyprinus carpio (common carp). The nanoparticle showed >100 mg/L for EC50 and LC50 against both aquatic organisms. Based on the result, it could be studied further to develop the ecofriendly formulation with stability studies for agriculture pest management.

Potential effect of Allium sativum bulb for the treatment of biofilm forming clinical pathogens recovered from periodontal and dental caries.

Biofilm producing clinical bacterial isolates were isolated from periodontal and dental caries samples and identified as, Lactobacillus acidophilus, Streptococcus sanguis, S. salivarius, S. mutansand Staphylococcus aureus. Among the identified bacterial species, S. aureus and S. mutansshowed strong biofilm producing capacity. The other isolated bacteria, Streptococcus sanguis, S. salivarius showed moderate biofilm formation. These pathogens were subjected for the production of extracellular polysaccharides (EPS) in nutrient broth medium and the strain S. aureus synthesized more amounts of EPS (610 ± 11.2 µg/ml) than S. sanguis (480 ± 5.8 µg/ml).EPS production was found to be less in S. salivarius (52 ± 3.8 µg/ml).The solvent extract of A. sativum bulb showed the phytochemicals such as, carbohydrate, total protein, alkaloids, saponins, flavonoids, tannins and sterioids. The solvent extract of A. sativum bulb showed wide ranges of activity against the selected dental pathogens. The difference in antibacterial activity of the solvent extract revealed differences in solubility of phytochemicals in organic solvents. Ethanol extract was highly active againstS. aureus (25 ± 2 mm). The Minimum Inhibitory Concentration (MIC) of crude garlic bulb varied widely and this clearly showed that bacteria exhibits different level of susceptibility to secondary metabolites. MIC value ranged between 20 ± 2 mg/ml and 120 ± 6 mg/ml and Minimum Bactericidal Concentration (MBC) value ranged from 60 ± 5 mg/l to 215 ± 7 mg/ml. To conclude, A. sativum bulb can be effectively used to treat periodontal and dental caries infections.

Prophylactic efficacy of Boerhavia diffusa L. aqueous extract in toluene induced reproductive and developmental toxicity in Drosophila melanogaster.

BACKGROUND: Environmental exposure to toxicants poses high risk to develop reproductive and developmental chronic toxicity in man. Toluene is one of the commonest industrial agents whose exposure is attributed with potential to induce reproductive and developmental toxicity. Since they contaminate the immediate environment of air and water to which humans are exposed, its containment is of great public health importance. Conventional treatment modalities fail owing to the difficulty to detect these highly volatile agents in environment and human body. The peril of such hazardous exposures is evident only when irreversible structural and functional damages have incurred. In such instances, prevention gains an upper hand when compared to therapeutic interventions. Several natural compounds derived from medicinal herbs possess potential to curb toxicities induced by such xenobiotic agents. Among them Boerhavia diffusa Linn. is a widely distributed and common herb attributed with antitoxic potential and capability for antioxidant defence. A study was performed on the prophylactic efficacy of aqueous extract of B. diffusa in curbing toluene induced developmental toxicity in Drosophila melanogaster. METHODS: The study consisted of a preliminary phytochemical screening and HPTLC profiling of B. diffusa aqueous extract (BDAE). LC50 of toluene was assessed and a sublethal dose of 200ppm was fixed for the study. Four doses of BDAE; 25, 50, 100 and 200mg/ml designated as Low dose, medium dose 1, medium dose 2 and high dose was used for the study. The parameters used for the study included the determination of larval period, pupal period, percentage of egg hatching, morphometric analysis of egg, larvae, pupae and adults, fertility, fecundity, lifespan and levels of antioxidant enzymes such as catalase, glutathione-S-transferase and superoxide dismutase. RESULTS: The phytochemical and HPTLC characters were as per the pharmacopoeial standards. LC50 of toluene was found to be 430ppm in this study. BDAE at medium dose 2 and high dose significantly prevented the deterioration of reproductive and developmental toxicity parameters of larval period, pupal period, percentage of egg hatching, morphometric characters of larva, pupa and adult, fertility, fecundity and lifespan in drosophila. Also the drug significantly elevated the levels of antioxidant enzymes. CONCLUSION: Toluene exposure during lifetime is inevitable. B. diffusa, equipped with its rich active ingredients prevented toluene induced developmental and reproductive toxicity in Drosophila. This medicinal herb provides a ray of hope in preventing environmental toxin induced reproductive and developmental toxicity.

One step green synthesis of larvicidal, and azo dye degrading antibacterial nanoparticles by response surface methodology.

The present study explored the one step extracellular green synthesis of Iron oxide (FexOy) and manganese oxide nanoparticles (MnNPs) using aqueous extract of Acorus calamus rhizome. The organic chemicals including polyphenol compounds responsible for bio-reduction and stabilization from the polyphenol enriched microwave irradiated aqueous extract of Acorus calamus were studied using GC-MS analysis. Further, their synthesis conditions were optimized using response surface methodology (RSM) and central composite design (CCD) using three variables. The green synthesized Iron oxide and Manganese oxide NPs were characterized by UV, FTIR, XRD, TEM and SEM. Results indicated that the Iron oxide NPs and mixture of iron and manganese NPs showed photocatalytic excellent activities in reducing dyes like methylene blue (0.1%) and Congo red (0.25%) at 0.03% NPs. However, Mn NPs showed moderate activity. On a contrary, manganese showed better larvicidal activity compared to Iron oxide NPs against the phytopathogens commonly affecting the vegetable crops. The present finding showed that high mortality rate at 30 µg/ml concentration of manganese NPs was comparatively interesting. In addition, NPs overall had appreciable activity with P. aeruginosa being more sensitive to Iron oxide NPs (22 ±â€¯2 mm zone of inhibition) and manganese NPs (13 ±â€¯2 mm zone of inhibition) and Iron oxide NPs completely inhibited the growth of A. flavus at 40 µg/ml concentration.

Green synthesis of silver nanoparticles using Rheum palmatum root extract and their antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa.

In the present study, silver nanoparticles (AgNPs) were synthesized by using aqueous root extracts of Rheum palmatum and characterized by various spectroscopic methods. The nanoparticles were found to be in hexagonal and spherical shapes. The average particle size was found to be 121 ± 2 nm with zeta potential values of -21.6 mv by dynamic light scattering (DLS) method. Gas chromatography-mass spectrometry (GC-MS) analysis of R. palmatum revealed 35 compounds. The synthesized AgNPs showed significant activity against Staphylococcus aureus and Pseudomonas aeruginosa with IC90 values of 15 µg/ml and IC50 values of 7.5 µg/ml, respectively. The protein leakage level was high and morphological changes occurred in bacteria treated with AgNPs.

Biosynthesized silver nanoparticles using floral extract of Chrysanthemum indicum L.--potential for malaria vector control.

Mosquitoes transmit serious human diseases, causing millions of deaths every year. The use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides synthesized of natural products for vector control have been a priority in this area. In the present study, silver nanoparticles (Ag NPs) were green-synthesized using a floral extract of Chrysanthemum indicum screened for larvicidal and pupicidal activity against the first to fourth instar larvae and pupae of the malaria vector Anopheles stephensi mosquitoes. The synthesized Ag NPs were characterized by using UV-vis absorption, X-ray diffraction, transmission electron microscopy, and energy-dispersive X-ray spectroscopy techniques. The textures of the yielded Ag NPs were found to be spherical and polydispersed with a mean size in the range of 25-59 nm. Larvae and pupae were exposed to various concentrations of aqueous extract of C. indicum and synthesized Ag NPs for 24 h, and the maximum mortality was observed from the synthesized Ag NPs against the vector A. stephensi (LC50 = 5.07, 10.35, 14.19, 22.81, and 35.05 ppm; LC90 = 29.18, 47.15, 65.53, 87.96, and 115.05 ppm). These results suggest that the synthesized Ag NPs have the potential to be used as an ideal eco-friendly approach for the control of A. stephensi. Additionally, this study provides the larvicidal and pupicidal properties of green-synthesized Ag NPs with the floral extract of C. indicum against vector mosquito species from the geographical location of India.

In-vitro antimicrobial, antibiofilm, cytotoxic, antifeedant and larvicidal properties of novel quinone isolated from Aegle marmelos (Linn.) Correa.

BACKGROUND: Plant metabolites have wide applications and have the potential to cure different diseases caused by microorganisms. The aim of the study was to evaluate the antimicrobial, antibiofilm, cytotoxic, antifeedant and larvicidal properties of novel quinine isolated from Aegle marmelos (Linn.) Correa. METHODS: A compound was obtained by eluting the crude extract, using varying concentrations of the solvents by the chromatographic purification. Broth micro dilution method was used to assess the antimicrobial activity and anticancer study was evaluated using MTT assay. Larvicidal activity was studied using leaf disc no-choice method. RESULTS: Based on the IR, 13C NMR and 1H NMR spectral data, the compounds were identified as quinone related antibiotic. It exhibited significant activity against Gram positive and Gram negative bacteria. The lowest Minimum Inhibitory Concentration (MIC) of the compound against Bacillus subtilis and Staphylococcus aureus was 100 and 75 µg mL(-1) respectively. Against Escherichia coli and Pseudomonas aeruginosa it exhibited MIC value of 25 µg mL(-1). The MIC of the compound against Aspergillus niger, A. clavatus, Penicillium roqueforti was 20 µg mL(-1) and that against Fusarium oxysporum (20 µg mL(-1)), A. oryzae (40 µg mL(-1)), and Candida albicans (60 µg mL(-1)), respectively. It showed effective antibiofilm activity against E. coli, S. typhii and P. aeroginosa at 8 µg mL(-1) and did not exhibit considerable cytotoxic activity against Vero and HEP2 cell lines. Additionally, the compound documented significant antifeedant and larvicidal activities against Helicoverpa armigera and Spodoptera litura at 125, 250, 500 and 1000 ppm concentrations. CONCLUSION: The results concluded that the compound can be evaluated further in industrial applications and also an agent to prepare botanical new pesticide formulations.

Rapid green synthesis of silver nanoparticles from Chrysanthemum indicum L and its antibacterial and cytotoxic effects: an in vitro study.

The present work reports a simple, cost-effective, and ecofriendly method for the synthesis of silver nanoparticles (AgNPs) using Chrysanthemum indicum and its antibacterial and cytotoxic effects. The formation of AgNPs was confirmed by color change, and it was further characterized by ultraviolet-visible spectroscopy (435 nm). The phytochemical screening of C. indicum revealed the presence of flavonoids, terpenoids, and glycosides, suggesting that these compounds act as reducing and stabilizing agents. The crystalline nature of the synthesized particles was confirmed by X-ray diffraction, as they exhibited face-centered cubic symmetry. The size and morphology of the particles were characterized by transmission electron microscopy, which showed spherical shapes and sizes that ranged between 37.71-71.99 nm. Energy-dispersive X-ray spectroscopy documented the presence of silver. The antimicrobial effect of the synthesized AgNPs revealed a significant effect against the bacteria Klebsiella pneumonia, Escherichia coli, and Pseudomonas aeruginosa. Additionally, cytotoxic assays showed no toxicity of AgNPs toward 3T3 mouse embryo fibroblast cells (25 µg/mL); hence, these particles were safe to use.

Characterization of ambrette seed oil and its mode of action in bacteria.

In the present study, chemical composition and the antibacterial mechanism of ambrette seed oil are investigated. Chemical composition of the oil was analysed by gas chromatography-mass spectrometry (GC-MS). Thirty-five compounds were identified and the major compounds were found to be farnesol acetate (51.45%) and ambrettolide (12.96%). The antibacterial activity was performed by well diffusion assay and the mechanisms were studied by measuring the alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and protein leakage assays. The antibacterial effect of the ambrette seed oil showed inhibitory effect against Bacillus subtilis, Staphylococcus aureus and Enterococcus faecalis. The LDH activity was high in all tested bacteria compared with control, whereas the ALP and protein concentrations were also increased in E. faecalis. Molecular docking revealed the ligands farnesol acetate and ambrettolide had satisfactory binding energy towards the beta lactamase TEM-72 and dihydrofolate reductase (DHFR) protein. Due to its better antibacterial properties, the ambrette seed oil could be used as a source of antibacterial agents.