Modulation of STAT3 signaling, cell redox defenses and cell cycle checkpoints by β-caryophyllene in cholangiocarcinoma cells: possible mechanisms accounting for doxorubicin chemosensitization and chemoprevention

Cholangiocarcinoma (CCA) is an aggressive group of biliary tract cancers, characterized by late diagnosis, low effective chemotherapies, multidrug resistance, and poor outcomes. In the attempt to identify new therapeutic strategies for CCA, we studied the antiproliferative activity of a combination between doxorubicin and the natural sesquiterpene β-caryophyllene in cholangiocarcinoma Mz-ChA-1 cells and nonmalignant H69 cholangiocytes, under both long-term and metronomic schedules. The modulation of STAT3 signaling, oxidative stress, DNA damage response, cell cycle progression and apoptosis was investigated as possible mechanisms of action. β-caryophyllene was able to synergize the cytotoxicity of low dose doxorubicin in Mz-ChA-1 cells, while producing cytoprotective effects in H69 cholangiocytes, mainly after a long-term exposure of 24 h. The mechanistic analysis highlighted that the sesquiterpene induced a cell cycle arrest in G2/M phase along with the doxorubicin-induced accumulation in S phase, reduced the γH2AX and GSH levels without affecting GSSG. ROS amount was partly lowered by the combination in Mz-ChA-1 cells, while increased in H69 cells. A lowered expression of doxorubicin-induced STAT3 activation was found in the presence of β-caryophyllene in both cancer and normal cholangiocytes. These networking effects resulted in an increased apoptosis rate in Mz-ChA-1 cells, despite a lowering in H69 cholangiocytes. This evidence highlighted a possible role of STAT3 as a final effector of a complex network regulated by β-caryophyllene, which leads to an enhanced doxorubicin-sensitivity of cholangiocarcinoma cells and a lowered chemotherapy toxicity in nonmalignant cholangiocytes, thus strengthening the interest for this natural sesquiterpene as a dual-acting chemosensitizing and chemopreventive agent

Solubility of Camphene and Caryophyllene Oxide in Subcritical and Supercritical Carbon Dioxide

The solubility of camphene and caryophyllene oxide in subcritical and supercritical carbon dioxide were determined using dynamic off-line analysis method. Under subcritical condition, solubility of camphene and caryophyllene oxide were (1.738 to 18.118) ∙ 10^-3 and (1.84 to 7.872) ∙ 10^-3 respectively at 298.15 K with pressure varied from 50 to 70 bar. At 302.15 K and under same pressure variation, solubility of camphene and caryophyllene oxide were (1.918 to 18.76) ∙ 10^-3 and (14 to 25.624) ∙ 10^-3 respectively. Under supercritical condition, experiments were run from 80 bar to 250 bar. Solubility of camphene was ranged from (54.024 to 151.67) ∙ 10^-3 at 308.15 K and (17.552 to 65.487) ∙ 10^-3 at 313.15 K; while solubility of caryophyllene oxide was ranged from (24.9 to 299.94) ∙ 10^-3 at 308.15 K and (2.542 to 102.359) ∙ 10^-3 at 318.15 K. These solubility data was correlated with three semi-empirical models which were Bartle, Chrastil and Mendez-Santiago-Teja model. Of these three models, Mendez-Santiago-Teja model showed excellent fitting with average absolute relative deviation kept below 2%

Chemical composition and insecticidal activity of plant essential oils from Benin against Anopheles gambiae (Giles)

BACKGROUND: Insecticide resistance in sub-Saharan Africa and especially in Benin is a major public health issue hindering the control of the malaria vectors. Each Anopheles species has developed a resistance to one or several classes of the insecticides currently in use in the field. Therefore, it is urgent to find alternative compounds to conquer the vector. In this study, the efficacies of essential oils of nine plant species, which are traditionally used to avoid mosquito bites in Benin, were investigated. METHODS: Essential oils of nine plant species were extracted by hydrodistillation, and their chemical compositions were identified by GC-MS. These oils were tested on susceptible “kisumu” and resistant “ladji-Cotonou” strains of Anopheles gambiae, following WHO test procedures for insecticide resistance monitoring in malaria vector mosquitoes. RESULTS: Different chemical compositions were obtained from the essential oils of the plant species. The major constituents identified were as follows: neral and geranial for Cymbopogon citratus, Z-carveol, E-p-mentha-1(7),8-dien-2-ol and E-p-mentha-2,8-dienol for Cymbopogon giganteus, piperitone for Cymbopogon schoenanthus, citronellal and citronellol for Eucalyptus citriodora, p-cymene, caryophyllene oxide and spathulenol for Eucalyptus tereticornis, 3-tetradecanone for Cochlospermum tinctorium and Cochlospermum planchonii, methyl salicylate for Securidaca longepedunculata and ascaridole for Chenopodium ambrosioides. The diagnostic dose was 0.77% for C. citratus, 2.80% for E. tereticornis, 3.37% for E. citriodora, 4.26% for C. ambrosioides, 5.48% for C. schoenanthus and 7.36% for C. giganteus. The highest diagnostic doses were obtained with S. longepedunculata (9.84%), C. tinctorium (11.56%) and C. planchonii (15.22%), compared to permethrin 0.75%. A. gambiae cotonou, which is resistant to pyrethroids, showed significant tolerance to essential oils from C. tinctorium and S. longepedunculata as expected but was highly susceptible to all the other essential oils at the diagnostic dose. CONCLUSIONS: C. citratus, E. tereticornis, E. citriodora, C. ambrosioides and C. schoenanthus are potential promising plant sources for alternative compounds to pyrethroids, for the control of the Anopheles malaria vector in Benin. The efficacy of their essential oils is possibly based on their chemical compositions in which major and/or minor compounds have reported insecticidal activities on various pests and disease vectors such as Anopheles

Enantiomeric Ratio Changes of Terpenes in Essential Oils from Hybrid Eucalyptus grandis × E. tereticornis and its Pure Species

Some Eucalyptus species produce oils with biological activities and the effect of their interspecifc hybridization on the enantiomeric composition of terpenes has not been reported. The enantiomeric excesses of monoterpenes in the essential oil of Eucalyptus grandis × E. tereticornis and its parental taxa were determined by gas chromatography-mass spectrometry (GC-MS), and to resolve coelutions problems by preparative high performance liquid chromatography (HPLC)and GC-MS with two columns in series. The essential oil composition of the hybrid presented qualitative and quantitative differences with the composition of its parental taxa. Great differences were found for the enantiomeric ratio in monoterpene alcohols among the three essential oils. Our results suggest that the enantiomeric analysis can be a reliable method for the study of how theinterspecifc hybridization can module the enantiomeric chemical profle in Eucalyptus essential oils. These results suggest the use of interspecifc hybridization to improve or expand the source of bioactive compounds.Fil: Naspi, Cecilia Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; ArgentinaFil: Alvarez Costa, Agustin. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; ArgentinaFil: Lucia, Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; ArgentinaFil: Gonzalez Audino, Paola Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; ArgentinaFil: Masuh, Hector Mario. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentin

Essential Oil Composition of Two Grammosciadium DC Species, G. platycarpum (Boiss et Hausskn) Schischk and G. macrodon Boiss (Apiaceae), from Turkey

Purpose: To investigate and compare the essential oil composition of two Grammosciadium species obtained by hydrodistillationMethods: The essential oil of the aerial parts of two species was obtained by hydrodistillation and analyzed by gas chromatography (GC) and gas chromatography - mass spectrometry (GC-MS).Results: In G. platycarpum, twenty-seven compounds were identified representing 99.3 % of the total oil. The major constituents of the oil were limonene (28.4 %), β-pinene (16.1 %), β-selinene (15.9 %), δ- 3-carene (11.2 %), α-farnesene (6.0 %) and p - cymene (4.7 %). The oil of G. macrodon contained thirty-seven components representing 94.4 % of the total oil, with caryophyllene oxide (15.9 %), germacrene - D (12.4 %), β-caryophyllene (11.5 %), δ-3 - carene (10.1 %) and cyclohexane (4.6 %) as the main constituents.Conclusion: The essential oil composition of G. macrodon was analysed for the first time, the most abundant constituents being caryophyllene oxide, and cyclohexane.Keywords: Grammosciadium macrodon, Grammosciadium platycarpum, Essential oil, Limonene, Caryophyllene oxide, Germacrene - D, β-Caryophyllene, δ-3-Caren

Potentiation of low-dose doxorubicin cytotoxicity by affecting p-glycoprotein through caryophyllane sesquiterpenes in hepg2 cells: an in vitro and in silico study

Doxorubicin represents a valuable choice for different cancers, although the severe side effects occurring at the high effective dose limits its clinical use. In the present study, potential strategies to potentiate low-dose doxorubicin efficacy, including a metronomic schedule, characterized by a short and repeated exposure to the anticancer drug, and the combination with the natural chemosensitizing sesquiterpenes β-caryophyllene and β-caryophyllene oxide, were assessed in human hepatoma HepG2 cells. The involvement of P-glycoprotein (P-gp) in the HepG2- chemosensitization to doxorubicin was evaluated. Also, the direct interaction of caryophyllene sesquiterpenes with P-gp was characterized by molecular docking and dynamic simulation studies. A metronomic schedule allowed us to enhance the low-dose doxorubicin cytotoxicity and the combination with caryophyllane sesquiterpenes further potentiated this effect. Also, an increased intracellular accumulation of doxorubicin and rhodamine 123 induced by caryophyllane sesquiterpenes was found, thus suggesting their interference with P-gp function. A lowered expression of P-gp induced by the combinations, with respect to doxorubicin alone, was observed too. Docking studies found that the binding site of caryophyllane sesquiterpene was next to the ATP binding domain of P-gp and that β-caryophyllene possessed the stronger binding affinity and higher inhibition potential calculated by MM-PBSA. Present findings strengthen our hypothesis about the potential chemosensitizing power of caryophyllane sesquiterpenes and suggest that combining a chemosensitizer and a metronomic schedule can represent a suitable strategy to overcome drawbacks of doxorubicin chemotherapy while exploiting its powerful activity

Chemopreventive potential of caryophyllane sesquiterpenes: an overview of preliminary evidence

Chemoprevention is referred to as a strategy to inhibit, suppress, or reverse tumor development and progression in healthy people along with high-risk subjects and oncologic patients through using pharmacological or natural substances. Numerous phytochemicals have been widely described in the literature to possess chemopreventive properties, although their clinical usefulness remains to be defined. Among them, caryophyllane sesquiterpenes are natural compounds widely occurring in nature kingdoms, especially in plants, fungi, and marine environments. Several structures, characterized by a common caryophyllane skeleton with further rearrangements, have been identified, but those isolated from plant essential oils, including β-caryophyllene, β-caryophyllene oxide, α-humulene, and isocaryophyllene, have attracted the greatest pharmacological attention. Emerging evidence has outlined a complex polypharmacological profile of caryophyllane sesquiterpenes characterized by blocking, suppressing, chemosensitizing, and cytoprotective properties, which suggests a possible usefulness of these natural substances in cancer chemoprevention for both preventive and adjuvant purposes. In the present review, the scientific knowledge about the chemopreventive properties of caryophyllane sesquiterpenes and the mechanisms involved have been collected and discussed; moreover, possible structure-activity relationships have been highlighted. Although further high-quality studies are required, the promising preclinical findings and the safe pharmacological profile encourage further studies to define a clinical usefulness of caryophyllane sesquiterpenes in primary, secondary, or tertiary chemoprevention

Influence of extraction methods on the composition of essential oils of Achillea millefolium L. from Lithuania

In this study, flowering aerial parts of Achillea millefolium were used as a matrix for supercritical CO2 extraction (SFE) of volatile oil. The collected extracts were analyzed by GC-FID and GC-MS methods and their composition were compared with that of the essential oil isolated by hydrodistillation (HD). The composition of the essential oil obtained by hydrodistillation and SFE methods is widely different. Indeed, the SFE volatile oil had a pale yellow color whereas the HD oil had a blue color due to the presence of chamazulene (48.0% vs. 4.3%). Other important constituents of HD oil were (E)- caryophyllene (19.5 %) and γ-muurolene (13.1%). The CO2 supercritical extract was dominated by (E)-caryophyllene (26.0%), γ-muurolene (22.0%), and caryophyllene oxide (8.1%)