RESUMEN
Bakuchiol (BAK) has been reported to have a diverse pharmacological property as an antibiotic, anti-cancer, anti-hypolipidemic, anti-inflammatory and anti-convulsant agent. This study aimed to elucidate the immunomodulation and anti-inflammatory mechanism of bakuchiol using lipopolysaccharide stimulated RAW 264.7 macrophages and various animal models. The present study has shown that BAK significantly suppressed the pro-inflammatory cytokine expression in a dose-dependent manner and its oral administration significantly decreased delayed hypersensitivity responses as compared to control group. The assessment of immunomodulatory activity was carried out by the testing Hemagglutinating antibody (HA) titer, delayed type hypersensitivity (DTH) responses and phagocytic index by carbon clearance test. On the other hand, it showed significant decrease in circulating antibody titer and carbon clearance assay in a concentration-dependent manner. BAK has significantly potentiated the cellular immunity as well as humoral immunity by facilitating the footpad thickness responses in sheep RBCs in sensitized mice by significantly decreasing circulating antibody titer. Molecular studies revealed that BAK inhibited the activation of upstream mediator nuclear factor-κB by suppressing the phosphorylation of IκBα and p65. The responses were statistically significant as compared with the control (*p < 0.05, **p < 0.01).
Asunto(s)
Antiinflamatorios/farmacología , Rechazo de Injerto/prevención & control , Hipersensibilidad Tardía/prevención & control , Inmunosupresores/farmacología , Inflamación/prevención & control , Macrófagos/efectos de los fármacos , Fenoles/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Rechazo de Injerto/inmunología , Rechazo de Injerto/metabolismo , Hipersensibilidad Tardía/inmunología , Hipersensibilidad Tardía/metabolismo , Inmunidad Humoral/efectos de los fármacos , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/metabolismo , Mediadores de Inflamación/metabolismo , Lipopolisacáridos , Activación de Linfocitos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , FN-kappa B/metabolismo , Fagocitosis/efectos de los fármacos , Fosforilación , Células RAW 264.7 , Ovinos , Transducción de Señal , Trasplante de PielRESUMEN
Despite substantial developments of extraction and separation techniques, isolation of natural products from natural sources is still a challenging task. Undoubtedly hybrid methods like liquid chromatography with NMR spectroscopy or liquid chromatography coupled with mass spectrometry made on-line structure elucidation possible and provided impressive examples of natural product identification without prior isolation, however, in many cases the necessity to get the purified compounds in hand is still a fact. The process begins with the collection of desired plant material which is subjected to the suitable extraction process. The complex crude extracts are then monitored by various chromatographic procedures to separate and quantify the desired compounds. The active plant extracts are then fractionated to isolate the bioactive compounds in their pure form. The fully identified compound is used as a lead for the production of related analogues to modulate the biological activity and to carry out structure-activity relationship. The major isolated bioactive compound is used for semi-synthetic modification or total synthesis should be carried out such that it is relatively easy to modify the structure of the lead compound. This is a simple and cost-effective way to increase the chance to discover lead compounds. The biological activity in vitro and in vivo has to be done after purification.
Asunto(s)
Productos Biológicos/análisis , Descubrimiento de Drogas/métodos , Cromatografía Liquida , Espectroscopía de Resonancia Magnética , Relación Estructura-ActividadRESUMEN
Betulinic acid (BA) is a pentacyclic triterpenoid natural product reported to inhibit cell growth in a variety of cancers. However, the further clinical development of BA got hampered because of poor solubility and pharmacological properties. Interestingly, this molecule offer several hotspots for structural modifications in order to address its associated issues. In our endeavor, we selected C-3 position for the desirable chemical modification in order to improve its cytotoxic and pharmacological potential and prepared a library of different triazoline derivatives of BA. Among them, we previously reported the identification of a potential molecule, that is, 3{1N(5-hydroxy-naphth-1yl)-1H-1,2,3-triazol-4yl}methyloxy betulinic acid (HBA) with significant inhibition of cancer cell growth and their properties. In the present study, we have shown for the first time that HBA decreased the expression of phosphotidylinositol-3 kinase (PI3K) p110α and p85α and caused significant downregulation of pAKT and of NFκB using human leukemia and breast cancer cells as in vitro models. Further it was revealed that PI3K inhibition by HBA induced cell cycle arrest via effects on different cell cycle regulatory proteins that include CDKis cyclins and pGSK3ß. Also, this target-specific inhibition was associated with mitochondrial apoptosis as was reflected by the increased expression of mitochondrial bax, downregulated bcl2 and decreased mitochondrial levels of cytochrome c, together with reactive oxygen species generation and decline in mitochondrial membrane potential. The apoptotic effectors such as caspase 8, caspase 9 and caspase 3 were found to be upregulated besides DNA repair-associated enzyme, that is, PARP cleavage caused cancer cell death. Pharmacodynamic evaluation revealed that both HBA and BA were safe upto the dose of 2000 mg/kg body weight and with acceptable pharmacodynamic parameters. The in vitro data corroborated with in vivo anticancer activity wherein Ehrlich solid tumor showed that HBA as a more potent agent than BA without any body weight loss and mortality.
Asunto(s)
Apoptosis/efectos de los fármacos , Puntos de Control del Ciclo Celular/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Neoplasias/enzimología , Fosfatidilinositol 3-Quinasas/metabolismo , Triterpenos/farmacología , Animales , Línea Celular Tumoral , Femenino , Humanos , Estructura Molecular , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Neoplasias/patología , Triterpenos Pentacíclicos , Fosfatidilinositol 3-Quinasas/genética , Inhibidores de las Quinasa Fosfoinosítidos-3 , Triterpenos/química , Ácido BetulínicoRESUMEN
The present study focused on the antibacterial and biofilm inhibitory potential of 4-epi-pimaric acid isolated from aerial parts (stem and leaves) of Aralia cachemirica L. (Araliaceae) against oral cavity pathogens. 4-epi-Pimaric acid exhibited minimum inhibitory concentration (MIC) in the range of 4-16 µg/ml and minimum bactericidal concentration (MBC) two- to four-folds higher than MIC. There was significant inhibition in the biofilm formation by Streptococcus mutans on the saliva coated surface (P < 0.05), and confocal microscopy revealed that 4-epi-pimaric acid inhibited the clumping and attachment of S. mutans. At 8 × MIC concentration, it significantly prevented the pH drop and reduced S. mutans biofilms (P < 0.05). Increased propidium iodide staining and leakage of 260- and 280-nm absorbing material by 4-epi-pimaric acid treated cells of S. mutans suggested that it probably causes disruption of the cytoplasmic membrane structure. It also exhibited significant suppression of TNF-α expression in human neutrophils, suggestive of its anti-inflammatory activity. Furthermore, the compound was found to be significantly safe (IC(50) >100 µg/ml) in the MTT assay on AML-12 cell lines. In conclusion, 4-epi-pimaric acid showed promising antibacterial, anti-biofilm and anti-inflammatory potency and this compound can be exploited for therapeutic application in oral microbial infections.
Asunto(s)
Antibacterianos/farmacología , Aralia/química , Biopelículas/efectos de los fármacos , Diterpenos/farmacología , Extractos Vegetales/farmacología , Streptococcus mutans/efectos de los fármacos , Animales , Antibacterianos/química , Antiinflamatorios/química , Antiinflamatorios/farmacología , Biopelículas/crecimiento & desarrollo , Línea Celular , Diterpenos/química , Diterpenos/toxicidad , Fibroblastos/efectos de los fármacos , Humanos , Ratones , Pruebas de Sensibilidad Microbiana , Boca/microbiología , Hojas de la Planta/química , Tallos de la Planta/química , Streptococcus mutans/crecimiento & desarrolloRESUMEN
Labisia pumila (Myrsinaceae), is a popular herb among the women in Malaysia known locally as "Kacip Fatimah". Recently many nutraceutical products containing the powdered or extracted parts of the plant have become available for women's health care. However no evaluation of the effect of the repeated dosing of any herbal product of this plant had been undertaken prior to a 28-day sub-acute study presented in this report. The results showed that a dose of 50mg/kg of an aqueous extract of L. pumila corresponded to no-adverse-effect-level (NOAEL), whereas higher doses were associated with some toxicity concerns.