Apis Mellifera Venom and Melittin Block neither NF-κB-p50-DNA Interactions nor the Activation of NF-κB, Instead They Activate the Transcription of Proinflammatory Genes and the Release of Reactive Oxygen Intermediates

Abstract

Many alternative treatment approaches, originating from Asia, are becoming increasingly popular in the Western hemisphere. Recently, an article published in a renowned journal reported that venom of apis mellifera (bee venom (BV)) and melittin mediate immune-modulating effects by blocking the activation of the transcription factor NF-κB. Such a modus operandi would corroborate the many claims of beneficial effects of BV treatment and give immediate credit to this form of therapy. Fibroblast-like synoviocytes from rheumatoid arthritis patients and dermal fibroblast cells and white blood cells from healthy volunteers were used to study the effects of BV and melittin on the activation of NF-κB and a series of genes that are markers of inflammation. EMSAs demonstrate that neither BV nor melittin blocked IL-1β-induced NF-κB activation; neither did they affect phosphorylation or degradation of IκB. Contrary to published data, even high concentrations of BV and melittin were without any effect on NF-κB-p50-DNA interactions. More importantly, in fibroblast-like synoviocytes, but also in dermal fibroblasts as well as in mononuclear cells exposed to BV or melittin, mRNA levels of several proinflammatory genes are significantly increased, and Western blot data show elevated cyclooxygenase-2 protein levels. Furthermore, exposure to BV higher than 10 μg/ml resulted in disintegration of all cell types tested. In addition, large quantities of oxygen radicals are produced in a dose-dependent manner in leukocytes exposed to BV. Taken together, data presented in this work do not corroborate an earlier report regarding the effectiveness of BV as an inhibitor of the transcription factor NF-κB.

Effects of Melittin on the Production of Matrix Metalloproteinase-1 and -3 in Rheumatoid Arthritic Fibroblast-Like Synoviocytes

“…Interestingly, we found that melittin does not inhibit TNF-α-or IL-1β-induced MMP3 expression, or NF-κB activity (data not shown). These results are consistent with a recent report that melittin does not decrease NF-κB activity induced by IL-1β or TNF-α (15). Taken together, these results suggest that melittin inhibits LPS-mediated FLS activation by acting on a signaling pathway that LPS does not share with IL-1β or TNF-α.…”

Evaluation of the Cytogenetic Status of Human Lymphocytes After Exposure to a High Concentration of Bee Venom In Vitro

“…This peptide has a highly cytotoxic effect on different types of cells, mostly due to disruption of the cell membrane. Namely, melittin shares its amphipathic properties with a series of peptides that affect the cell membrane bilayer integrity, either by creating defects, disrupting it, or by forming pores (44).…”

Hyaluronan Production in Synoviocytes as a Consequence of Viral Infections

“…EMSA-Preparation of nuclear extract, execution of EMSA, and EMSA specificity control experiments have been described in detail on a number of occasions (16,19,20). In short, oligonucleotides resembling consensus binding sites for NF-B (5Ј-AGT TGA GGG GAC TTT CCC AGG C-3Ј), AP-1 (c-Jun) (5Ј-CGC TTG ATG AGT CAG CCG GAA-3Ј), and CRE (5Ј-AGA GAT TGC CTG ACG TCA GAG AGC TAG-3Ј)) were purchased from Promega (Promega, Mannheim, Germany).…”

“…A number of additional controls (competition and supershift assays) for EMSAs utilizing the consensus NF-B elements have been shown before (17,19). Parts of such control experiments are also shown in Fig.…”