ABSTRACT
Propolis has been used to treat several diseases since ancient times, and is an important source of bioactive natural compounds and drug derivatives. These properties have kept the interest of investigators around the world, leading to the investigation of the chemical and biological properties and application of propolis. In this report, the chemical constituents that are responsible for the anticancer activities of propolis were analyzed. The propolis was sourced from Al-Baha in the southern part of the Kingdom of Saudi Arabia. Standard protocols for chemical fractionation and bioactivity-guided chemical analysis were used to identify the bio-active ethyl acetate fraction. The extraction was performed in methanol and then analyzed by gas chromatography-mass spectrometry (GC-MS). The major compounds are triterpenoids, with a relative concentration of 74.0%; steroids, with a relative concentration of 9.8%; and diterpenoids, with a relative concentration of 7.9%. The biological activity was characterized using different approaches and cell-based assays. Propolis was found to inhibit the proliferation of cancer cells in a concentration-dependent manner through apoptosis. Immunofluorescence staining with anti-α-tubulin antibodies and cell cycle analysis indicated that tubulin and/or microtubules are the cellular targets of the L-acetate fraction. This study demonstrates the importance of Saudi propolis as anti-cancer drug candidates.
Subject(s)
Propolis/chemistry , Propolis/pharmacology , Acetates/pharmacology , Alanine Transaminase/blood , Animals , Apoptosis/drug effects , Aspartate Aminotransferases/blood , Blood Glucose/metabolism , Cell Cycle Checkpoints/drug effects , Cell Line, Tumor , Cell Proliferation/drug effects , Cholesterol/blood , Cluster Analysis , Creatinine/blood , Electric Impedance , Gas Chromatography-Mass Spectrometry , Humans , Inhibitory Concentration 50 , Jurkat Cells , Kidney/drug effects , Kidney/pathology , Lipoproteins, HDL/blood , Liver/drug effects , Liver/pathology , Male , Microtubules/drug effects , Microtubules/metabolism , Rats , Saudi ArabiaABSTRACT
Propolis is a glue material collected by honeybees which is used to seal cracks in beehives and to protect the bee population from infections. Propolis resins have a long history in medicinal use as a natural remedy. The multiple biological properties are related to variations in their chemical compositions. Geographical settings and availability of plant sources are important factors for the occurrence of specific natural products in propolis. A propolis ethylacetate extract (800mg) from Saudi Arabia (Al-Baha region) was separated by preparative scale high-speed countercurrent chromatography (HSCCC) using a non-aqueous solvent system n-hexane-ACN (1:1, v/v). For multiple metabolite detection, the resulting HSCCC-fractions were sequentially injected off-line into an atmospheric pressure chemical ionization mass-spectrometry (APCI-MS/MS) device, and a reconstituted mass spectrometry profile of the preparative run was visualized by selected ion traces. Best ion-intensities for detected compounds were obtained in the negative APCI mode and monitored occurring co-elution effects. HSCCC and successive purification steps resulted in the isolation and characterization of various bioactive natural products such as (12E)- and (12Z)-communic acid, sandaracopimaric acid, (+)-ferruginol, (+)-totarol, and 3ß-acetoxy-19(29)-taraxasten-20a-ol using EI-, APCI-MS and 1D/2D-NMR. Cycloartenol-derivatives and triterpene acetates were isolated in mixtures and elucidated by EI-MS and 1D-NMR. Free fatty acids, and two labdane fatty acid esters were identified by APCI-MS/MS. In total 19 metabolites have been identified. The novel combination of HSCCC fractionation, and APCI-MS-target-guided molecular mass profiling improve efficiency of lead-structure identification.
Subject(s)
Propolis/chemistry , Acetates , Animals , Bees , Chemical Fractionation , Chromatography, High Pressure Liquid/methods , Countercurrent Distribution/methods , Molecular Weight , Propolis/metabolism , Saudi Arabia , Solvents , Stereoisomerism , Tandem Mass Spectrometry/methodsABSTRACT
Development of hypopharyngeal glands (HPG) of native honey bee workers (Apis mellifera jemenitica) and Carniolan hybrid bees reared under normal constipations was studied. The HPG development degree and acini surface were influenced with age. The development degrees increased gradually. The maximum developmental degree was recorded at 9-days old in both two races. The development decreased after 12 days. The HPG development degree and acini surface of all ages in native bees were significantly less than Carniolan hybrid bees. Histological studies on HPG showed that there were some differences between the glands of the two races at the maximum developmental stage (9-days). The staining of cell cytoplasm by haematoxylin and eosin was similar. However secretory cells numbers were more in Carniolan hybrid than the native one.
