Abstract
Bee venom (BV) from honey bee (Apis mellifera L.) has been used in oriental medicine and cosmetic ingredients because of its diverse pharmacological activities. In many studies, among BV components, phospholipase A2 (PLA2) is known as a major player in BV-induced allergic reaction. Therefore, we removed PLA2 from BV using ultrafiltration and then investigated in vitro phototoxicity and in vivo skin sensitization of PLA2-free BV (PBV) in comparison with regular BV. The 3T3 neutral red uptake phototoxicity assay can be appropriated to identify the phototoxic effect of a test substance upon the exposure of ultraviolet A. Chlorpromazine, a positive control, showed high levels of photoirritation factor and mean photo effect values, while BV and PBV had less of these values. Local lymph node assay is an alternative method to evaluate skin sensitization potential of chemicals. BALB/c mice were treated with p-phenylenediamine (PPD, positive control), BV, or PBV. In all of PPD concentrations, stimulation indexes (SI) as sensitizing potential of chemicals were ≥1.6, determined to be sensitizer, while SI levels of BV and PBV were below 1.6. Thus, based on these findings, we propose that both BV and PBV are nonphototoxic compounds and nonsensitizers.
Highlights
Bee venom (BV) from honey bee (Apis mellifera L.) is a complex mixture of active peptides and various enzymes, such as melittin, phospholipase A2 (PLA2), apamin, adolapin protease-inhibitors, bioactive amines, and mast cell degranulating peptide [1, 2]
The BALB/c 3T3 cells were exposed to various concentration of the test materials, after which they were exposed to UVA (5 J/cm2) for 1 h, following which the cell viability was determined as per the neutral red uptake (NRU) phototoxicity assay
Inflammation and allergic contact dermatitis are usually associated with various proinflammatory mediators, including nitric oxide (NO) and prostaglandin E2, which are the markers of inflammatory reactions production and the synthesis of inflammatory cytokines [20]
Summary
Bee venom (BV) from honey bee (Apis mellifera L.) is a complex mixture of active peptides and various enzymes, such as melittin, phospholipase A2 (PLA2), apamin, adolapin protease-inhibitors, bioactive amines, and mast cell degranulating peptide [1, 2]. Fibroblasts have lower toxicity and higher sensitivity than keratinocytes when exposed to UVA irradiation [9,10,11] Due to this reason, neutral red uptake (NRU) phototoxicity assay using BALB/c 3T3 cell line was adopted by the Organization for Economic Co-operation and Development (OECD) in 2004, as a Test Guideline (TG) [12]. LLNA-ELISA method does not require the use of radioisotope 3H-thymidine, but 5-bromo-2-deoxyuridine (BrdU), which is incorporated into the DNA of proliferating lymphocytes during the S-phase of cell cycle This assay has been approved by the Environmental Protection Agency (EPA), OECD, Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), and the European Centre for the Validation of Alternative Methods (ECVAM). Our studies revealed that PBV and BV can be developed for cosmetic application because they appear to have neither phototoxicity nor sensitizing effects
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