Photoprotective effects of Romanian propolis on skin of mice exposed to UVB irradiation

We aimed at investigating the antioxidant, antiinflamatory, antiapoptotic and antigenotoxic effects of a Romanian Propolis (RP) extract in two concentrations (RP1 3 mg, respectively RP2 1.5 mg polyphenols/cm²), topically administered, either prior to or after UVB exposure, in a Swiss mouse model. Our results showed that both concentrations of RP extract, independent of the time of administration, significantly attenuated the malondialdehyde (MDA) formation and restored glutathione peroxidase (GPx) activity. However, the 8-hydroxy-2-deoxy-guanosine (8-oxo-dG), nitric oxide (NO) and glutathione (GSH) levels were not influenced by UVB exposure and RP treatment. Interleukin (IL)-6 levels were significantly decreased by RP treatment, both before and after UVB-exposure. RP2 extract, in both regimens, significantly reduced the epidermal hyperplasia and dermal inflammation, whereas RP1 pre-treatment diminished only the dermal inflammation. The effect of our RP extract in terms of reduction of sunburn cell formation and of activated caspase-3 and TUNEL-positive cells was observed in both subsets of the experiment, RP2 having a slightly better protective effect as compared to RP1. The antigenotoxic effect of RP was demonstrated by significantly reduced cyclobutane pyrimidine dimers (CPDs) formation. Our results suggest that RP extract might be a potential chemopreventive candidate by modulation of multiple UVB-induced signaling pathways in skin.     

Comparative study of the antibacterial activity of propolis from different geographical and climatic zones

Propolis is a natural substance produced by honeybees upon collection and transformation of resins and exudates from plants. Comparative studies on propolis collected from a wide range of countries are crucial for linking its provenance to antibacterial activity and thus ensuring that the beneficial properties of propolis are used more efficiently by the general public. This study reports the in vitro screening of ethanol extracts of propolis (n = 40), collected from a wide range of countries within the tropical, subtropical and temperate zones, and on the comparison of their activity against a range of Gram-positive and Gram-negative bacteria using a broth microdilution assay. The results obtained revealed that propolis extracts were mostly active against Gram-positive bacteria. The samples were subjected to principal component analysis (PCA) in order to model their activity against Gram-positive microorganisms. Three distinct clusters were distinguished in the PCA mapping based on MIC values, categorizing samples with strong (MIC range 3.9-31.25 mg/L), moderate (MIC range 31.25-> or =500 mg/L) and weak antibacterial activity or inactivity (MIC > or = 500 mg/L only). It is hypothesized that for samples of tropical provenance differences in the activity profiles may depend on the climatic characteristics of the collection sites. High antibacterial activity was observed for samples from locations characterized by a wet-tropical rainforest-type climate.     

Proteomic analysis identifies differentially expressed proteins after red propolis treatment in Hep-2 cells

Here we investigated alterations in the protein profile of Hep-2 treated with red propolis using two-dimensional electrophoresis associated to mass spectrometry and apoptotic rates of cells treated with and without red propolis extracts through TUNEL and Annexin-V assays. A total of 325 spots were manually excised from the two-dimensional gel electrophoresis and 177 proteins were identified using LC–MS–MS. Among all proteins identified that presented differential expression, most were down-regulated in presence of red propolis extract at a concentration of 120 μg/mL (IC50): GRP78, PRDX2, LDHB, VIM and TUBA1A. Only two up-regulated proteins were identified in this study in the non-cytotoxic (6 μg/mL) red propolis treated group: RPLP0 and RAD23B. TUNEL staining assay showed a markedly increase in the mid- to late-stage apoptosis of Hep-2 cells induced by red propolis at concentrations of 60 and 120 μg/mL when compared with non-treated cells. The increase of late apoptosis was confirmed by in situ Annexin-V analysis in which red propolis extract induced late apoptosis in a dose-dependent manner. The differences in tumor cell protein profiles warrant further investigations including isolation of major bioactive compounds of red propolis in different cell lines using proteomics and molecular tests to validate the protein expression here observed.     

蜂胶醇提物通过抑制C/EBP同源蛋白表达减轻氧化低密度脂蛋白诱导的血管内皮细胞凋亡

目的:研究蜂胶醇提物(ethanol extract of propolis,EEP)对氧化低密度脂蛋白(oxidized low-density lipoprotein,ox-LDL)诱导的血管内皮细胞凋亡的抑制作用,并探讨可能的分子机制。方法:体外培养人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs),给予EEP(7.5、15和30 mg/L)、4-苯丁酸(4-phenylbutyric acid,PBA;4 mmol/L)预处理1 h,再加入ox-LDL(100 mg/L)或衣霉素(tunicamycin,TM;4 mg/L)继续培养24 h。分别采用MTT法和Annexin V-FITC/PI双染法检测细胞活力和凋亡情况;试剂盒测定培养液乳酸脱氢酶(lactic dehydrogenase,LDH)和细胞内caspase-3活性。分别采用Western blot和real-time PCR技术检测内质网应激(endoplasmic reticulum stress,ERS)凋亡途径关键蛋白C/EBP同源蛋白(C/EBP homologous protein,CHOP)和Bcl-2的表达变化。结果:与ERS抑制剂PBA相似,EEP呈剂量依赖性地减轻ox-LDL所诱导的HUVECs损伤,表现为细胞活力增加(P0.01或P0.05),LDH漏出、凋亡率和caspase-3活性降低(P0.05或P0.01),且可抑制ERS诱导剂TM所致的HUVECs活力下降(P0.05),以及LDH漏出、细胞凋亡率和caspase-3活性增加(P0.05或P0.01);与PBA相似,EEP可抑制ox-LDL所诱导的CHOP上调和Bcl-2下调(P0.05或P0.01);另外,与TM组比较,EEP预处理组CHOP蛋白和mRNA表达上调也受到明显抑制(P0.05或P0.01)。结论:EEP可减轻oxLDL所诱导的HUVECs凋亡,其机制可能与抑制CHOP介导的ERS凋亡途径有关。     

Phloroglucinols from Anti‐Microbial Deposit‐Resins of Australian Stingless Bees (Tetragonula carbonaria)

Stingless bees accumulate deposits of plant resins that are mixed with beeswax to produce propolis. Previous studies have reported anti‐microbial constituents of stingless bee (Tetragonula carbonaria) propolis from East Australia, but several components remained to be characterized. In the search of natural products yet unreported for Australian propolis, four bee deposit‐resins of T. carbonaria bees were analysed by gas and liquid chromatography mass spectrometry with accurate mass measurements. Ethanolic extracts of the deposit‐resins were tested in vitro against Staphylococcus aureus ATCC 25983 and Pseudomonas aeruginosa ATCC 27853 by the agar diffusion method. Phloroglucinols, flavonoids and isoprenoids were identified in samples. The crude extracts showed strong anti‐staphylococcal effects but were less active against the Gram‐negative bacterium. The diagnostic data enabled the identification of markers that can be used for profiling other Australian propolis sources and to target the isolation of bioactive phloroglucinols in future studies against antibiotic resistant S. aureus strains. Copyright © 2014 John Wiley & Sons, Ltd.