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Bee Venom: Its Potential Use in Alternative Medicine
1Laboratory of Research on Local Animal Products, Ibn-Kh aldoun University of Tiaret, 14000, Alge-
ria; 2Department of Biology, Faculty of Life and Nature Sciences, Mohamed El Bachir El Ibrahimi
University, Bordj Bou Arreridj, 34000, Algeria
Abstract: In recent times, bee venom (BV) from honey bee (Apis mellifera L.) has become the focus
of interest as a form of alternative and preventive medicine for the treatment of a number of clinical
cases such as arthritis, rheumatism, pain, cancer and a vast range of other conditions. BV contains sev-
eral biochemically and pharmacologically active substances. Some of these compounds are well stud-
ied and th eir mechanisms of action established, despite the fact that few are undergoing clinical trials.
Scientists are now performing intensive research work especially human clinical trials to improve the
potential use of BV and its chemical constituents as the next drugs generation in the treatment of chronic disorders. N ever-
theless, the dual effect of some bee venom components is also impo rtant in the design of future therapeutic goals. This pa-
per gives recent evidences on the chemical and biological properties of the major components of bee venom, their under-
lying molecular action mechanism, and reasons of its consideration as a promising alternative medicine.
Keywords: Alternative medicine, Bee venom, biological properties, clinical trials, chronic disorders, molecular mechanism.
Many products based on traditional knowledge are im-
portant sources of income, food and health care for large
populations throughout the world. Since ancient times, tradi-
tional medicines and natural products are being used for the
treatment of many disorders. Bee venom from honey bee
(Apis mellifera L.), also known as apitoxin, has traditionally
been used in folk medicine to relieve pain and to treat in-
flammatory diseases such as arthritis and rheumatism [1-4].
BV still continues to be used in a similar way until the cur-
rent time. The first scientific work on BV was published in
the late 19th century where rheumatic patients of the Austrian
physician, Philip Terc, were improved following bee stings
treatment . Undoubtedly, BV is in no way a new d iscov-
ery since its use dated back to ancient times more than 6000
years (medicine in ancient Egypt). The Greeks and Romans
also used bee products for medicinal purposes. Hippocrates
(460-370 BC), Aristotle (384-332 BC) and Galen (130-200
AD), prescribed the use of honey and bee venom as a cure
for baldness .
Bee venom has been shown to contain several biochemi-
cally or pharmacologically active substances including poly-
peptides (melittin, apamin, and mast cell degranulating pep-
tide), amines (histamine, serotonin, dopamine, and norepi-
nephrine), and enzymes (phospholipase, hyaluronidase, his-
tidine decarboxylase) [7-10]. These substances were claimed
to directly or indirectly express its potency and medical effi-
cacy. Bee venom has been suggested as an effective healing
agent for alleviating persistent pain and treating several
*Address correspondence to this author at the Department of Biology, Fac-
ulty of Life and Nature Sciences, Mohamed El Bachir El Ibrahimi Univer-
sity, Bordj Bou Arreridj, 34000, Algeria; Tel: +213 34 293 442;
E-mails: firstname.lastname@example.org; email@example.com
ailments including different rheumatic disorders involving
inflammation and degeneration of connective tissue (differ-
ent types of arthritis) [1, 11-13], neurological disorder ,
related-immune syndrome (multiple sclerosis)  and der-
matological conditions (eczema, psoriasis, herpes virus in-
fections) . BV was revealed to be effective in the reduc-
tion of tumors of many different types of malignant d iseases
[15, 16], as it was found to stimulate natural immunity
through activation of the pituitary and adrenal glands and to
stimulate the body to produce natural cortisone . Un-
questionably, BV has become the focus of interest as a form
of alternative and preventive medicine. Scientific works are
especially focusing on human clinical trials to improve the
potential use of BV and its chemical constituents as the next
drugs generation in the treatment of chronic disorders. This
review aims to summarize the evidence to date elucidating
the most salient biological properties of BV. It also revealed
prominent knowledge on the responsible underlying action
mechanism of bee venom constituents and how BV and/or its
components support its potential use as an alternative therapy.
BIOACTIVE COMPONENTS OF BEE VENOM
Among the several bee products, BV has been high-
lighted due to its chemical composition . BV is toxic,
contains 88 % water and only 0.1µg dry venom. The dry
venom is known to be a very complex mixture of peptides
including melittin, apamin, adolapin, and the MCD peptide.
It also contains enzymes (e.g., PLA2), biologically active
amines (e.g., histamine and epinephrine) and nonpeptide
components (including lipids, carbohydrates and free amino
acids. Table 1 summarizes the major components of bee
venom, their biochemical characteristics and their main
therapeutical and pharmacological properties.
2 Anti-Infective Agents, 2015, Vol. 13, No. 1 Yuva Bellik
Table 1. Components of bee venom and their major biological properties [4, 19-21].
(% dry weight)
Strong surface activity on cell lipid membrane
Anti-inflammatory in very small doses Activation of PLA2
Increases capillary permeability and blood circulation; lowers blood pres-
sure and blood coagulation, exhibits immuno-stimulatory and immuno-
Radiation protective; influences the central nervous system.
Exhibits anticancer, antibacterial, antifungal and antiviral properties
Exhibits anti-inflammatory properties; stimulates the release of cortisone;
exhibits antiserotonin action
Cytotoxic effect against cancer
Inhibits Ca2+-activated K+ channel
Possesses immuno-suppressor and nociceptive effect
Exhibits anti-inflammatory, analgesic effect and nociceptive effect
Inhibits histamine release in low dose and high dose
Shows anti-allergic effect
Exhibits anti-inflam matory a ctivity , anti-nociceptive, and antipyretic effect
Inhibits of PLA2 and COX activity
Inhibits of erythrocytes aggregation
Procamine A, B
Inhibits the activity of different proteases like trypsin,
chymotprypsin, plasmin and thrombin, thus decreasing inflammation and
High-affinity binding sites in rat brain
High-affinity binding sites in rat brain
Inhibition of phosphodiesterase and the enzyme-activating c apacity o f
β-Adrenergic and anti-arrhythmic effects
Phosph olipase A2
Exhibits antigenicity, allergenicity, inflammatory activity and nociceptive
Prevents neuronal cell death caused by prion peptides.
Phosph olipase B
Exhibits nerve regeneration, activation of neurons and glial cells, facilita-
tion of neurotransmitter release and interaction with melittin
Exhibits detoxicating activity; cleavage of the toxic lysolecetin
Selectively attacks tissue hyaluronic acid polymers
Increases the permeability of capillaries
Dilates blood vessels, increases the permeability of capillaries and in-
creases blood circulation
Stimulates smooth muscles
Agonist at dopamine receptors
Agonist at adrenoceptors
Causes alarm for the bee colony and its defensive behavior
P, Ca, Mg
Bee Venom: Its Potential Use in Alternative Medicine Anti-Infective Agents, 2015, Vol. 13, No. 1 3
Melittin, a 26-residue peptide , present as the prevail-
ing toxic component of the BV, accounts for 50% of its
composition . The sequence of melittin is Gly-Ile-Gly-
Ile-Ser-Trp-Ile-Lys-Arg-Lys-Arg-Gln-Gln , in which the
amino-terminal region (residues 1–20) is predominantly hy-
drophobic with no lytic activity whereas the carboxy-
terminal region (residues 21–26) is hydrophilic and respon-
sible for the lytic action . The amphipathic property of
melittin makes it water-soluble as a monomer or as a
tetramer [26-28]. This polypeptide readily inserts into mem-
branes and disrupts both natural and synthetic phospholipid
bilayers  (Fig. 1). In fact, the affinity of melittin for
membranes, composed of negatively charged lipids, has been
shown to be about 100-fold greater than for zwitterionic lip-
ids . The action of melittin in membranes is mediated
through pore formation, wh ich causes membrane permeation
 and lyses both prokaryotic and eukaryotic cells in a non-
selective manner . This mode of action is responsible for
hemolytic , anti-microbial [34, 35], anti-fungal ,
anti-tumor  and leishmanicidal  activities of melittin .
Melittin also enhances the activity of PLA2 by the mediation
of enhanced influx of calcium ions . In addition, melittin
is a potent inhibitor of Na+K+-ATPase , and Ca2+-
ATPase  as well as H+K+-ATPase . Melittin exerts
several other biological properties such as inhibiting replica-
tion of HIV-1 by suppressing viral gene expression; melittin
reduces levels of intracellular Gag and viral mRNAs, and
decreases HIV long terminal repeat (LTR) activity , anti-
transformation effect by specifically eliminating cells that
express the oncoprotein , and mediation of signal trans-
duction through stimulating nucleotide exchange by hetero-
trimeric GTP-binding proteins . Alternatively, erythro-
cyte membrane represents the prime target for the action of
melittin; the latter binds rapidly to erythrocytes, reduces the
rotational mobility of band 3 protein in human erythrocyte,
and induces the release of hemog lobin .
The structural and pharmacological properties of apamin
might have a central role in its cytotoxic effects on cancer
cells and its nociceptive activity . Apamin is a small bee
venom polypeptide that corresponds to less than 2% of
venom dry weight and consists of 18 amino acids containing
2 disulfide bridges. Arg-13 and Arg-14 are in the active site
of the toxin . It is the only polypeptide neurotoxin that
passes the blood-brain barrier and induces hyperexcitability
. Apamin blocks the Ca2+-activated K+ channels  in
several cells and does not present lytic properties. As for
other potent venom neurotoxins, apamin binds with high
affinity to specific receptors of a post-synaptic membrane
and blocks many inhibitory or hyper-polarization effects,
including α-adrenergic, cholinergic, purinerg ic, and relaxes
the neurotensin-induced effects .
Adolapin is a basic polypeptide with 103 amino acids
residues and comprising 1% of dry bee venom . It has
been reported to possess anti-nociceptive, anti-inflammatory
and antipyretic effects . Adolapin can inhibit prostaglan-
din synthesis via inhibition of cyclooxygenase activity .
It also inhibits the lipoxygenase from human platelets. In
addition, adolapin has been shown to have an analgesic ef-
The mast cell degranulation peptide (MCDP) is chemi-
cally similar to apamin and accounts for 22 amino acid resi-
dues. It is rich in α -helix, and presents two disulfide bonds
in its structure between Cys3,15 and Cys5,19. The sequence of
Fig. (1). Melittin makes channels into phospholipids vesicle membranes.
4 Anti-Infective Agents, 2015, Vol. 13, No. 1 Yuva Bellik
MCDP is lle-Lys-Cys-Asn-Cys-Lys-Arg-His-Val-lle-Lys-
Pro-His-lle-Cys-Arg-Lys-lle-Cys-Gly-Lys-Asn . This
peptide possesses striking immunological and pharmacologi-
cal activities. At higher concentrations, it has anti-
inflammatory properties, but at low concentrations, it is a
strong mediator of mast cell degranulation and histamine
release . It was suggested that at high doses, disulfide
exchange between IgE and the MCDP on the mast cell sur-
face might inhibit the release of histamine, which would al-
low the MCDP to act as an anti-allergic agent . MCDP is
also an epileptogenic neurotoxin, an avid blocker of the po-
tassium channels and can cause a significant lowering of the
blood pressure in rats .
Bee venom phospholipase A2 (BV-PLA2) is the most
lethal bee venom peptide. It consists of a single chain of 128
amino acid residues and contains four disulphide bridges
. BV-PLA2 comprises 12-15% of the dry weight of bee
venom and it is extremely alkaline. It is a hydrolytic enzyme
that specifically cleaves the sn-2 acyl bond of phospholipids
at the lipid/water interface . Its activity can be enhanced
by melittin. Many studies have reported a synergistic action
of BV-PLA2 with melittin during erythrocyte lysis process
. It has been demonstrated that melittin helps in the ex-
posure of membrane phospholipids to the catalytic site of the
enzyme, by opening melittin-induced channels . BV-
PLA2 exerts many other pharmacological activities including
anti-human immunodeficiency virus (HIV) activity, neuro-
toxicity , myotoxicity, and neurite out growth induction [58,
Hyaluronidase is the enzyme responsible for hyaluronic
acid hydrolysis and condroitin sulfate, and to a small extent,
dermatan sulfate. Hyaluronidase isolated from bee venom, a
potent allergen originally described as a "spreading factor",
specifically degrades hyaluronic acid in the extracellular
matrix of skin, thereby facilitating the diffusion of other
venom constituents into the body. Hyaluronidase from bee
venom is a single polypeptide composed of 350 residues. It
is secreted as a basic glycoprotein with a carbohydrate that
accounts for 7% the protein mass , and contains two di-
sulfide bridges and four potential glycosylation sites.
Hyaluronidase shares a 50% sequence identity with
hyaluronidases from other hymenoptera and, also with sev-
eral mammalian enzymes (as much as 30 % identity) .
In fact, bee venom contains an arsenal of biologically
active molecules, in addition to those aforementioned, it of-
fers a great hope as an important source in the development
of potential therapeutic agents and their application for treat-
ing comp lex diseases.
COLLECTION OF BEE VENOM
Collecting bee venom requires careful work with the
highest degree of cleanliness, since the venom is injected
directly without further processing or sterilization. The
venom collector must ensure safety against the disturbed
bees to obtain dry venom. Early collection method of bee
venom required surgical removal of the venom gland or
squeezing each individual bee until a droplet could be col-
lected from the tip of the sting . Since the early 1960's,
extraction by the electro-shock method has continuously
improved and is at present the standard procedure . Dif-
ferent extraction or collection methods result in different
compositions of the final products. Venom co llected under
water to avoid evaporation of very volatile compounds
seems to yield the most potent venom . Venom collected
from surgically removed venom sacs showed different pro-
tein contents from that collected with the electroshock
method . The standard electro-shock method cannot be
recommended for venom collection from African honeybees
or the more defensive races . Colony arousal can become
so overwhelming that bees start killing each other and alert
other colonies or attack the beekeeper and bystanders. The
mass reaction of African honeybees may also result in con-
tamination of the collected venom. Nevertheless, venom is
collected by this method in Brazil and Argentina, with only
minor modifications. Even European colonies remain dis-
turbed for up to a week after collection and it is stated by
Mitev  that colonies from which venom has been col-
lected, every three days they produce 14% less honey.
Meanwhile, Morse and Benton  found no such evidence
for reduced productivity. Galuszka  found that when
using electro-shock treatment, the most efficient collection
cycle was three 15-minute stimulations at intervals of three
days, repeated after 2-3 weeks. An Argentinean beekeeper
found that by modifying the electric stimulus, his collection
efficiency greatly increased and the bees remained disturbed
for less time .
Various trap designs stimulate bees by applying a mild
electric shock through wires above the collecting tray. The
most widely used designs are modifications of the one first
presented by Benton and coworkers . A review by Mraz
 discusses further developments. The trays are placed
either between the bottom board and brood chamber at the
hive entrance or in a special box between supers and the hive
cover . It is unlikely that a bee will eject all the contents
of its venom sac, even after repeated stinging . There-
fore, typically, only 0.5 to 1.0 jil (0.2 j£l) of venom can be
collected per bee, with an average of ten stings per bee .
This results in less than 0.1 ijg (0.11 jig) of dry venom per
bee. Consequently, at least 1 million stings are required to
make one gram of dry bee venom. Dotimas and Hider 
reported that 1 g of venom can be collected from twenty
hives over a two hour period. Exact production figure is un-
available. The main venom producer in the USA had pro-
duced about 3000 grams of venom over 30 years .
BEE VENOM: AN ALTERNATIVE MEDECINE
The use of bee products as a medicine dates back to the
most ancient written records. They have continued to be used
in modern folk medicine ever since. In addition to the other
bee products, bee venom and its chemical components have
been actively studied in many countries including the United
States, China, Russia, and Northern European countries and
received scientific endorsement. Evidences suggested that
BV can be useful in a wide variety of medical situations such
as arthritis and rheumatism, pain and cancer. The literature
of bee venom therapy is very extensive. Below are described
most studied and well-known BV constituents including
Bee Venom: Its Potential Use in Alternative Medicine Anti-Infective Agents, 2015, Vol. 13, No. 1 5
their major biological properties as well as their mode of
Anti-inflammatory and Anti-nociceptive Properties
Different cytokines are extremely associated with in-
flammatory disorders, especially, tumor necrosis factor-α
(TNF-α) and inter leukin-1β (IL-1β), which are major con-
tributors to chronic inflammatory diseases . In recent
years, many of the inflammatory disorders are becoming
frequent in aging society. Rheumatoid arthritis and os-
teoarthritis are the major inflammatory disorders affecting
people worldwide. Unfortunately, the current clinically used
anti-inflammatory medications have the inconveniences of
adverse effects, partial efficacy and high cost of drug .
This has highlighted the need for safer and more effective
treatments [74, 75]. Accordingly, the discovery of drugs that
can be used for the treatment of inflammatory diseases is
important in human health . Bee venom anti-
inflammatory activity is one of the most extensively investi-
gated biological actions [12, 77-81]. Many research works
have demonstrated that bee venom injection can reduce the
inflammation process in arthritis and rheumatism [1, 13, 19,
82-84]. Bee venom (0.5, 1, and 5 µg/ml) and melittin (5 and
10 µg/ml) were reported to decrease LPS-induced production
of PGE2, TNF-α, and sodium nitroprusside (SNP)-induced
NF-κB activation by preventing p50 translocation through
interaction of melittin and sulfhydryl residue of p50 and/or
IκB kinases (IKKα and IKKβ) [80, 85]. Son and colleagues
 proposed the anti-arthritic mechanism of BV in prevent-
ing RA (Fig. 2). In addition, BV inhibited the production of
nitric oxide (NO), expression of COX-2, inducible NOS
(iNOS), and cytosolic PLA2 (cPLA2) [86, 87]. The decrease
in COX-2 and PLA2 expression, and the decrease in the lev-
els of TNF-α, IL-1, IL-6, NO, ROS, and intracellular calcium
were reported to be associated with the anti-inflammatory
effect. Moreover, the inhibition of PLA2 activity by BV has
an important role in suppressing the progression of RA. Re-
ports from clinical trials have showed that bee venom acu-
puncture (BVA) treatment was very effective in arthritic
patients . Likewise, patients treated with BVA therapy
twice a week for 3 months showed a significant decrease in
tender joint, swollen joint and duration of morning stiffness
. In addition, BVA was clinically effective in knee arthri-
tis patien ts . Collective evidence from in vitro and in
vivo experiments showed that BVA may become a promising
treatment for both rheumatoid arthritis [3, 83] and os-
Recently, it has been reported that some calcium channel
blockers can decrease areas of atherosclerotic lesions, ROS
production and expression of inflammatory cytokines .
Apamin has long been known as a highly selective blocker
of Ca2+-activated K+ channels . Apamin exhibited anti-
inflammatory effects on serotonin and dextran oedemas pro-
voked in the rat paw. It also inhibited the elevation of the
haptoglobin and seromucoid content in the sera of rats and
haptoglobin content in the sera of rabbits with model in-
flammation . Apamin (0.05$mg/kg) reduced expression
of TNF-α, lipids, Ca2+ levels, vascular cell adhesion mole-
cule (VCAM)-1, intracellular cell adhesion molecule
(ICAM)-1, TGF-β1, and fibronectin as well as the nuclear
factor kappa B (NF-κB) signaling pathway [94, 95]. Another
study showed that the anti-atherosclerotic effect of apamin is
displayed by decreasing the apoptotic macrophages through
reducing the expression of pro-apoptotic genes (Bax,
caspase-3 and poly-(ADP-ribose) polymerase (PARP) pro-
tein) levels, as well as through increasing expression of anti-
apoptotic genes (Bcl-2 and Bcl-xL) . Likewise, BV in-
hibited the proliferation of rheumatoid synovial cells by in-
ducing apoptosis through decreasing the expression of B-cell
leukaemia/lymphoma-2 (BCL2) and the activation of BCL2-
associated X protein (BAX) and caspase-3 .
The MCD peptide, a strong mast cell degranulating factor
from bee venom, has potent anti-inflammatory activity (at
doses as low as 0.1 mg/kg) in a variety of animal models
. It was proposed that the peptide might be a useful
therapeutic agent in some arthritic conditions . As for the
anti-inflammatory (anti-arthritis) effects of adolapin, it is
presumably due to its ability to inhibit the PG synthesis sys-
Interestingly, other pre-clinical studies on several animal
models have demonstrated that bee venom can be used not
only as an anti-inflammatory therapeutic agent but also for
safe anti-nociceptive purposes [12, 73, 98-100]. In addition,
BV injection can cause an initial nociceptive action as well
as a prolonged anti-nociceptive effect. Following the subcu-
taneous injection of melittin, MCD peptide and PLA
related, it has been noticed that the three peptides were able
to produce distinct nociceptive paw flinches through the re-
lease of proinflammatory or inflammatory mediators and/or
endogenous algogenic substances . Among the studied
polypeptides, melittin was found to be responsible for the
prolonged painful stimulation of BV injection, leading to
both tonic nociception and hypersensitivity, while the other
polypeptides contributed only to the early nociceptive re-
sponses. Recent evidences in rodents have highlighted the
action of ERKs, p38 and JNK in generating nociceptive sen-
sitivity and neural plasticity in the pain sensory system, par-
ticularly in the dorsal horn of the spinal cord [102-104]. Bee
venom induced nociceptive processing through direct and
indirect actions of BV components. Nociceptors can be acti-
vated through activation of membrane-bound pain sensors
that cause an increase of intracellular Ca2+ concentration and
phosphorylation of various subtypes of MAPKs and PKs
(PKC and PKA), that hypersensitize ionic nociceptor mole-
cules (TRPV1, P2X3, ASICs, and 5-HT3) and G-protein
coupled receptors (P2Y1, H1, BK1 and BK2), leading to up-
regulation of TTX-resistant voltage-dependent sodium chan-
nels (Fig. 3) .
Bee venom has been reported to exhibit anti-tumor activ-
ity both in vitro and in vivo . One of the most impli-
cated facts in tumorigenesis and anti-tumor therapy is apop-
tosis, a process of central importan ce implicated in the
pathogenesis and pathophysiology of several human diseases
. Apoptotic cell death process is regulated by the ex-
pression of several proteins. The main involved are members
of bcl-2 family  and cysteine proteases caspases .
Nonetheless, evidences from various studies have stated that
apoptosis, necrosis and lysis of tumor cells were proposed as
possible mechanisms by which BV inhibited tumor growth
6 Anti-Infective Agents, 2015, Vol. 13, No. 1 Yuva Bellik
Fig. (2). Proposed anti-arthritic effect of BV (melittin). BV (melittin) inhibits the release of IκB through the inhibition of IKKs. This inhibi-
tion might be due to an interaction between the sulfhydryl (SH) group of IKKα and IKKβ with BV (melittin) molecule, which results in
NF-κB inactivation, and thus reduces the generation of inflammatory mediators. BV (melittin) may also interact directly with p50 of NF-κB
and thereby inhibits the translocation of p50 into the nucleus.
Abbreviations: P, phosphorus; Ub, ubiquitin; NF-κB, nuclear factor-κB; IκB, inhibitor of NF-κB; IKK, IκB kinase; NEMO, NF-κB essential
[15, 16, 109-111]. Many reports have shown that BV can
induce cell cycle arrest, growth inhibition, and apoptosis in
various tumor cells including ovary, renal, lung, liver, pros-
trate, bladder, mammary and cancer leukemia cells as well as
breast cells. Bee venom was reported to exert anti-tumor
activity through induction of apoptosis and inhibition of
COX-2 mRNA expression and PGE2 synthesis in human
lung cancer NCI-H1299 cells . A more recent study
evidenced that BV (1–5 µg/ml) inhibited the growth of lung
cancer cells by induction of apoptosis through increase of
death receptor 3 expression and inactivation of NF-κB in
lung cancer cell lines A549 and NCI-H460 . BV (10
µg/ml) also induced apoptosis of human RA proliferated
synovial fibroblasts through decrease in BcL2 expression and
an increase in Bax and caspase-3 expression . Likewise,
it has been demonstrated that BV induced apoptosis in hu-
man leukemic U937 cells through downregulation of the
ERK and Akt signal pathway, which is followed by down-
regulation of Bcl-2, activation of caspase-3, p38 MAPK and
JNK . BV has been shown to induce apoptosis in hu-
man osteosarcoma MG-63 cells , and mammary carci-
noma cell proliferation in vitro and tumor growth in vivo
. BV and melittin (approximately 0.4-0.8 µg/ml) inhib-
ited the proliferation of v ascular smooth muscle cells
through induction of apoptosis via suppression of NF-ƙB and
Akt activation, and downregulation of Bcl-2 . It (1.0,
3.0, 9.0 mg/kg) also inhibited K1735M2 mouse melanoma
cells in vitro and the growth of murine B16 melanomas in
vivo . BV treatment protected against ethanol-induced
hepatocyte apoptosis through the regulation of Bcl-2 with the
subsequent inactivation of the caspase and poly-(ADP-
ribose) polymerase (PARP) . Recent study on structure-
activity relationship showed that melittin can initiate an
apoptotic machinery that depends on calcium influx and ac-
tivation of Ca2+/calmodulin-dependent protein kinase
(CaMKII), transforming growth factor-β-activated kinase 1
(TAK1), and JNK/p38 signaling pathway. In addition, melit-
tin can sensitize hepatocellular carcinoma cells to tumor ne-
crosis factor-related apoptosis-inducing ligand (TRAIL-
induced apoptosis) by activating CaMKII-TAK1-JNK/ p38
and inhibiting IKK-NFƙB pathways . Moreover, melittin
could inhibit the growth and angiogenesis of human hepato-
cellular carcinoma BEL-7402 cell [117, 118]. BV also in-
duced cytotoxic effects on TSGH-8301 human bladder can-
cer cells through intracellular Ca2+-modulated intrinsic death
pathway . Furthermore, melittin-loaded perfluorocar-
bon nanoparticles possessed the ability to safely deliver im-
portant payloads via intravenous ways, which target and kill
tumor cells .
Bee Venom: Its Potential Use in Alternative Medicine Anti-Infective Agents, 2015, Vol. 13, No. 1 7
Fig. (3). A schematic drawing of proposed underlying mechanisms of the bee venom induced nociception and hyperalgesia to heat and me-
chanical stimuli applied in the periphery. melittin, MCD pept ide, apamin and tertiapin bind directly to the membrane of a nociceptor cell lead-
ing to activation of it. Meanwhile, melittin, MCD peptide, bv PLA2, and hyaluronidase cause tissue damage and release ATP and H+ that
activate P2X3/P2Y, TRPV1 and ASIC. Indirect actions of melittin, MCD peptide and bv PLA2 cause degranulation of mast cells and release
histamine, BK and 5-HT that activate H1 receptor, 5-HT3 receptor and BK1/2 receptors. The firing of nociceptor terminals will be mediated
by voltage-dependent sodium channels (TTXr Nav1.8/1.9), VDCC, VDPC, Kir and Ca2+–K+. Dorsal root reflex and axon reflex may cause
release of glutamate and neuropeptides (SP and CGRP) that further activate their autoreceptors on the nociceptor terminals or blood vessels
causing inflammatory extravasation (neurogenic) with infiltration of macrophage, immune cells and platelets and many cytokines (TNFalpha,
IL1beta, PAF, etc.). The syringe indicates transcutaneous injection of bee venom.
Abbreviations: 5-HT3, 5-hydroxytryptamine receptor 3; 12-HETE, 12-hydroxyeicosatetraenoic acids; AA, arachidonic acid; ASIC, acid-
sensing ionic channel; ATP, adenosine triphosphate; BK1/2, bradykinin receptors 1/2; bv PLA2, bee venom phospholipase A2; Ca2+–K+,
calcium-dependent potassium channel; CGRP, calcitonin-gene related peptide; COX-1/2, cyclooxygenases1/2; Glu, glutamate; H1, histamine
receptor type 1; iGluRs, ionotropic glutamate receptors; IL1β, interleukin 1β; IL6, interleukin 6; Kir, inward-rectifier potassium channel,
LOXs, lipoxygenases; MAPKs, mitogen-activated protein kinases; MCD peptide, mast cell degranulating peptide; MCL peptide, mastocy-
tolyitic peptide; NK1, neurokinin 1; NOS, notric oxide synthase; P2X3, P2-purinoreceptor X3; P2Y, P2- purinoreceptor Y; PAF, platelet-
activated factor; PGs, prostaglandins; PKA, protein kinase A; protein kinase C; protein kinase G; SP, substance P; TNFα, tumor-necrosis
factor α; TRPV1, transient receptor potential vanilloid receptor 1; TTXr, tetrodotoxin-resistant; VDCC, voltage dependent calcium channel,
VDPC, voltage-dependent potassium channel .
Interestingly, it has been shown recently that BV induced
cell cycle arrest and apoptosis both in human breast cancer
MCF7 cells  and human cervical epidermoid carcinoma
Ca Ski cells  via a mitochondria-dependent pathway.
More recently, it has been demonstr ated that BV
(1-10$µg/ml) and melittin (0.5-2.5$µg/ml) inhibited prostate
cancer cells in vitro and in vivo through induction of apop-
totic cell death mediated by the suppression of constitutively
activated NF-κB . The molecular mechanism underly-
ing apoptotic effect of BV is now well established and de-
scribed. Several authors have proposed the mode of action of
bee venom and/or melittin in anti-tumor th erapy depending
on the cancer cell type [119, 121-124].
Multiple Sclerosis Improvement
Recently, there has been considerable interest in the psy-
choneurological approach of bee venom therapy (BVT), es-
pecially in the context of its mode of action as neuroprotec-
tive agents in progressive neurodegenerative disorders such
as multiple sclerosis, Parkinson, and Alzheimer diseases.
Since multiple sclerosis (MS) is a chronic neurological dis-
order characterized by inflammation, demyelination, and
8 Anti-Infective Agents, 2015, Vol. 13, No. 1 Yuva Bellik
axonal degeneration in the central nervous system (CNS)
that has a negative consequential impact on the lives of pa-
tients and their families, MS can cause motor, sensory, or
visual impairment; fatigue; bowel, bladder, and sexual dys-
function; cognitive impairment; and depression . Re-
cently, many surveys have showed that people with MS, in
addition to the conventional medications, frequently used
complementary and alternative medicine treatments includ-
ing bee venom therapy [126-130]. Nonetheless, bee sting
therapy has been used by a small number of MS patien ts
. Almost no animal or human substantial studies have
so far showed significant effect of BV for patients with pro-
gressive forms of MS. The most randomized investigations
conducted on MS patients have demonstrated that no signifi-
cant reduction in the number of lesions, relapse rate, disabil-
ity, fatigue, and quality of life was detected after application
of the bee sting therapy [131-133], despite the highly anti-
inflammatory properties of BV constituents including melit-
tin and adolapin. Authors deemed that the inefficiency of BV
in the improvement of MS was due to the PLA2 present in
BV. As it was reported that PLA2 is expressed in abundance
in the lesions of MS animal model , thus, inhibiting
PLA2 may prevent both the onset and the progression of MS
. Others believed that BV may be an effective treatment
for patients with multiple sclerosis. An extensive study has
been carried out by Krivopalov-Moskvin and colleagues
(http://www.api-centre.ru). They concluded that over 2000
MS patients, only 5-7% of MS patients showed no improve-
ment following the BVT. In addition, Hauser and coworkers
 revealed improvement rates between 50 to 60 %.
Other Biological Properties of BV
Antibacterial and Antifungal
Bee venom has long been known to have a n atural antim-
icrobial effect [136-139]. The antibacterial properties of BV
are due to the potential action of melittin , which has
very low cell selectivity and acts strongly on the cell mem-
brane lipid through pores forming channels. BV has been
referred to as the natural penicillin  and has been re-
ported to kill both gram-positive and gram-negative bacteria
[140, 141-143]. It has been also shown that melittin com-
pletely dissolved gram-positive as well as gram-negative
bacteria . Recently, a comparative antifungal study re-
vealed that BV exhibited prominent antifungal activities
Interestingly, Nermine and colleagues  assessed the
immunological effects of BV in mice with intracerebral can-
didiasis and demonstrated that BV enhanced the host’s im-
mune response, manifested by significant decrease in the
fungal load in the brain and significant increase in TNF-α m-
RNA expression. Park and Lee  found that melittin ex-
erted its antifungal effect via apoptosis.
It is worthnoting however, that one of the most widely
known and extensively tested properties of bee products is
the antimicrobial activity of honey. Many scientific tests
have been conducted with a variety of bacteria, fungi, viruses
and other microorganisms. Table 2 summarizes the suscepti-
bility of the most frequently studied and well-known bacte-
rial strains to honey especially manuka honey. It should be
noted that other reports demonstrating similar results are not
Oxidative stress is considered to play a pivotal role in the
pathogenesis of aging and several degenerative diseases.
Studies have highlighted the ability of BV (0, 1, 2.5, 5 or 10
µg/kg) to remove the deleterious effects of reactive oxygen
species (ROS) . BV potently inhibited the production of
-) and hydrogen peroxide in human neutrophil
. In addition, it was assumed that BV acupuncture de-
creased the level of ROS induced oxidative injury to syno-
vial fluid proteins . Reports from animal and human ex-
periments revealed that BV may protect against sequel of
oxidative stress induced by rheumatoid arthritis [78, 80, 162,
164, 165]. Importantly, BV is endowed of radioprotective
effects [166-168] and protected against the deleterious ef-
fects of ionizing radiations in terms of reduction of chromo-
somes aberrations in bone marrow cells of Wistar rats in vivo
Bee venom immunotherapy (BVI) has been reported to
be highly valuable in healing, capable of improving health
related quality of life [170, 171]. BVI is effective for reduc-
ing local and systemic allergic reactions. It has been revealed
that up to 95% of people susceptible to bee sting w ere pro-
tected from the risk of systemic reactions . However,
BVI can assure a co mplete protection against adverse (aller-
gic) reactions from stings . In fact, BVI uses much
lower amounts of BV when compared to those used in treat-
ing arthritic patients. Sublingual immunotherapy with the
introduction of BV under the tongue is safe , and can
significantly reduce reactions in people allergic to bee stings
Although few research works were done on the antipara-
sitic properties of BV and BV constituents. Recently, an im-
portant genetic investigation demonstrated that expression of
the honey bee PLA2 gene in the midgut of transgenic mos-
quitoes severely reduced their ability to sustain plasmodium
development and the transmission of the parasite to other
vertebrate hosts . In addition, it has been shown that
cecropin A-melittin hybrid peptides show remarkable
leishmanicidal activity [177-179], which involves targeting
of the plasma membrane of Leishmania donavani promas-
tigote. Further, N-terminal fatty acylation has been shown to
increase the leishmanicidal activity of the hybrid peptides
BEE VENOM THERAPY LIMITATIONS
Although the promise of BV application in alternative
medicine for the evidences discussed above, the use of BV in
conventional medicine has languished, and the foregoing BV
therapy is often painful. Human and animal toxicity cases
have been shown for BV following bee stings [180, 181].
Understandably, there is no doubt that BV and its chemical
components are not without cytotoxic effects. Bee stings are
Bee Venom: Its Potential Use in Alternative Medicine Anti-Infective Agents, 2015, Vol. 13, No. 1 9
Table 2. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of honey against various bac-
Antimicrobial activity of
Antimicrobial activity of
[149, 150, 153]
[146, 147, 157]
indeed unsafe for allergic people. A recent study revealed
that about 1 to 5 % of the people worldw ide are hypersensi-
tive to BV or other insects like wasps and hornets . In
addition, it has been referred that both hyaluronidase and
phospholipase A2 are the main allergens since they can cause
pathogenic reactions in the majority of patients susceptible to
BV where it has been observed that 71% of patients had spe-
cific serum IgE to recombinant Hya and 78% to recombinant
phospholipase A2 . Moreover, it has been found that
constituents of, and whole bee venom cause cell membrane
instability, especially severe disruption of red blood cell
membranes and apoptosis and necrosis of several cell types.
Stuhlmeier  showed that BV and melittin considerably
raise mRNA levels of pro-inflammatory genes including
COX-2, TNF-α and IL-8 in dermal fibroblasts and mononu-
clear cells from healthy persons and cultured human fibro-
blast-like synoviocytes from rheumatoid arthritis patients.
Furthermore, it has been recognized that the nociceptive ef-
fect caused both in the peripheral and central nervous sys-
tems follow ing bee venom injection significantly increases
the risk of development of prolonged pathological pain con-
Bee venom has long been a remedy and considered to be
highly important traditional and alternative medicine for the
treatment of various illnesses. A significant number of in
vitro and in vivo as well as clinical studies provided substan-
tial evidences that BV and its chemical constituents are ef-
fective inhibitors of the inflammatory and several other
chronic processes. The mechanisms of the protective effects
may be several fold including effects on cellular growth (dif-
ferentiation or cell cycle), apoptosis, anti-inflammatory, anti-
nociceptive immunosuppressive effects, antitumor and anti-
cancer activities. The$inhibitory$ability$of$BV$and$ its$con4
and$its$components. Despite these considerations, there is a
resistance in Western medical circles either to accept these
results or to test bee venom treatments according to the
Western medical standards. In addition, the existing scien-
tific research works are replete with non-randomized and
uncontrolled investigations, and there is a real paucity of
well-intended human clinical trials evaluating the therapeutic
effectiveness of BV, as well as the lack of standard-
ized BV formulations and restricted protocols for BV appli-
cation. Accordingly, more experiments on single BV con-
stituents as well as whole bee venom are necessary. Further
detailed studies at cellular and molecular levels, appropriate
animal models and human clinical investigations are needed.
In view of the foregoing, future researches should ideally
10 Anti-Infective Agents, 2015, Vol. 13, No. 1 Yuva Bellik
conduct human intervention trials with large sample sizes
and adequate design to prove the clinical efficacy and safety
profile of BV, establish what, if any, adverse effects are ob-
served, and assess effects on relapse levels and disease pro-
gression. The goal of prospect BV therapy efforts will be
purely pharmacological aspects including several relevant
concerns such as standardized preparations, method for ad-
ministering BV, applications forms, dose, toxicity, bioavail-
ability and delivery system.
CONFLICT OF INTEREST
The author confirms that this article content has no con-
flict of interest.
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Received: October 09, 2014 Revised: Ja nua ry 28, 2015 Accepted: February 19, 2015
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