Melittin is a basic 26-amino-acid polypeptide that constitutes 40–60% of dry honeybee (Apis mellifera) venom. Although much is known about its strong surface activity on lipid membranes, less is known about its pain-producing effects in the nervous system. In this review, we provide lines of accumulating evidence to support the hypothesis that melittin is the major pain-producing substance of bee venom. At the psychophysical and behavioral levels, subcutaneous injection of melittin causes tonic pain sensation and pain-related behaviors in both humans and animals. At the cellular level, melittin activates primary nociceptor cells through direct and indirect effects. On one hand, melittin can selectively open thermal nociceptor transient receptor potential vanilloid receptor channels via phospholipase A2-lipoxygenase/cyclooxygenase metabolites, leading to depolarization of primary nociceptor cells. On the other hand, algogens and inflammatory/pro-inflammatory mediators released from the tissue matrix by melittin’s pore-forming effects can activate primary nociceptor cells through both ligand-gated receptor channels and the G-protein-coupled receptor-mediated opening of transient receptor potential canonical channels. Moreover, subcutaneous melittin up-regulates Nav1.8 and Nav1.9 subunits, resulting in the enhancement of tetrodotoxin-resistant Na+ currents and the generation of long-term action potential firing. These nociceptive responses in the periphery finally activate and sensitize the spinal dorsal horn pain-signaling neurons, resulting in spontaneous nociceptive paw flinches and pain hypersensitivity to thermal and mechanical stimuli. Taken together, it is concluded that melittin is the major pain-producing substance of bee venom, by which peripheral persistent pain and hyperalgesia (or allodynia), primary nociceptive neuronal sensitization, and CNS synaptic plasticity (or metaplasticity) can be readily induced and the molecular and cellular mechanisms underlying naturally-occurring venomous biotoxins can be experimentally unraveled.
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Habermann E. Bee and wasp venoms. Science 1972, 177: 314–322.
Son DJ, Lee JW, Lee YH, Song HS, Lee CK, Hong JT. Therapeutic application of anti-arthritis, pain-releasing, and anti-cancer effects of bee venom and its constituent compounds. Pharmacol Ther 2007, 115: 246–270.
Chen J, Lariviere WR. The nociceptive and anti-nociceptive effects of bee venom injection and therapy: A double-edged sword. Prog Neurobiol 2010, 92: 151–183.
Chen J, Guan SM. Bee venom and pain. In Toxinology: Toxins and Drug Discovery. Edited by Gopalakrishnakone P. New York: Springer; in press.
Williams JC, Bell RM. Membrane matrix disruption by melittin. Biochim Biophys Acta 1972, 288: 255–262.
Tosteson MT, Tosteson DC. The sting. Melittin forms channels in lipid bilayers. Biophys J 1981, 36: 109–116.
Tosteson MT, Alvarez O, Hubbell W, Bieganski RM, Attenbach C, Caporales LH, Levy JJ, Nutt RF, Rosenblatt M, Tosteson DC. Primary structure of peptides and ion channels. Role of amino acid side chains in voltage gating of melittin channels. Biophys J 1990, 58: 1367–1375.
Schwarz G, Zong RT, Popescu T. Kinetics of melittin induced pore formation in the membrane of lipid vesicles. Biochim Biophys Acta 1992, 1110: 97–104.
Fattal E, Nir S, Parente Jr RA, Szoka FC. Pore-forming peptides induce rapid phospholipid flip-flop in membranes. Biochemistry-US 1994, 33: 6721–6731.
Smith R, Separovic F, Milne TJ, Whittaker A, Bennett FM, Cornell BA, Makriyannis A. Structure and orientation of the pore-forming peptide, melittin, in lipid bilayers. J Mol Biol 1994; 241: 456–466.
Bechinger B. Structure and functions of channel-forming peptides: magainins, cecropins, melittin and alamethicin. J Membr Biol 1997, 156: 197–211.
Chen LY, Cheng CW, Lin JJ, Chen WY. Exploring the effect of cholesterol in lipid bilayer membrane on the melittin penetration mechanism. Anal Biochem 2007, 367: 49–55.
Chen X, Wang J, Kristalyn CB, Chen Z. Real-time structural investigation of a lipid bilayer during its interaction with melittin using sum frequency generation vibrational spectroscopy. Biophys J 2007, 93: 866–875.
Matsuzaki K, Yoneyama S, Miyajima K. Pore formation and translocation of melittin. Biophys J 1997, 73: 831–838.
Klocek G, Schulthess T, Shai Y, Seelig J. Thermodynamics of melittin binding to lipid bilayers: Aggregation and pore formation. Biochemistry 2009, 48: 2586-2596.
Mollay C, Kreil G. Enhancement of bee venom phospholipase A2 activity by melittin, direct lytic factor from cobra venom and polymyxin B. Febs Lett 1974, 46: 141–144.
Hassid A, Levine L. Stimulation of phospholipase activity and prostaglandin biosynthesis by melittin in cell culture and in vivo. Res Commun Chem Pathol Pharmacol 1977, 18: 507–517.
Nishiya T. Interaction of melittin and phospholipase A2 with a zobenzene containing phospholipid. J Biochem 1991, 109: 383–388.
Vernon LP, Bell JD. Membrane structure, toxins and phospholipase A2 activity. Pharmacol Ther 1992, 54: 269–295.
Sharma SV. Melittin-induced hyperactivation of phospholipase A2 activity and calcium influx in ras-transformed cells. Oncogene 1993; 8: 939–947.
Li KC, Chen J. Altered pain-related behaviors and spinal neuronal responses produced by s.c. injection of melittin in rats. Neuroscience 2004, 126: 753–762.
Chen YN, Li KC, Li Z, Zhang ZW, Ji YH, Gao GD, Chen J. Effects of bee venom peptidergic components on rat behaviors related to pain and inflammation. Neuroscience 2006, 138: 631–640.
Du YR, Xiao Y, Lu ZM, Ding J, Xie F, Fu H, Wang Y, Strong JA, Zhang JM, Chen J. Melittin activates TRPV1 receptors in primary nociceptive sensory neurons via the phospholipase A2 cascade pathways. Biochem Biophys Res Commun 2011, 408: 32–37.
McMahon SB, Koltzenburg M, Tracey I, Turk DC. Wall and Melzack’s Textbook of Pain. Philadelphia: Elsevier; 2006.
Schmidt RF, Willis WD. Encyclopedia of Pain. Berlin: Springer; 2007.
Chen J, Han JS, Zhao ZQ, Wei F, Hsieh JC, Bao L, Chen ACN, Dai Y, Fan BF, Gu JG, Hao SL, Hu SJ, Ji YH, Li YJ, Li YQ, Lin Q, Liu XG, Liu YQ, Lu Y, Luo F, Ma C, Qui YH, Rao ZR, Shi L, Shyu BC, Song XJ, Tang JS, Tao YX, Wan Y, Wang JS, et al. Pain. In Neuroscience in the 21 st Century: From Basic to Clinical. Edited by Pfaff DW. New York; Springer; 2013: 965–1023.
Koyama N, Hirata K, Hori K, Dan K, Yokota T. Computer-assisted infrared thermographic study of axon reflex induced by intradermal melittin. Pain 2000, 84: 133–139.
Koyama N, Hirata K, Hori K, Dan K, Yokota T. Biphasic vasomotor reflex responses of the hand skin following intradermal injection of melittin into the forearm skin. Eur J Pain 2002, 6: 447–453.
Sumikura H, Andersen OK, Drewes AM, Arendt-Nielsen L. A comparison of hyperalgesia and neurogenic inflammation induced by melittin and capsaicin in humans. Neurosci Lett 2003, 337: 147–150.
Sumikura H, Andersen OK, Drewes AM, Arendt-Nielsen L. Secondary heat hyperalgesia induced by melittin in humans. Eur J Pain 2006, 10: 121–125.
Schumacher MJ, Schmidt JO, Egen NB, Dillon KA. Biochemical variability of venoms from individual European and Africanized honeybees (Apis mellifera). J Allergy Clin Immunol 1992, 90: 59–65.
Schumacher MJ, Tveten MS, Egen NB. Rate and quantity of delivery of venom from honeybee stings. J Allergy Clin Immunol 1994, 93: 831–835.
Armstrong D, Dry RM, Keele CA, Markham JW. Method for studying chemical excitants of cutaneous pain in man. J Physiol 1951, 115: 59–60.
Armstrong D, Dry RM, Keele CA, Markham JW. Pain-producing substances in blister fluid and in serum. J Physiol 1952, 117: 4p–5p.
Armstrong D, Dry RM, Keele CA, Markham JW. Pain-producing actions of tryptamine and 5-hydroxytryptamine. J Physiol 1952, 117: 70P–71P.
Armstrong D, Dry RM, Keele CA, Markham JW. Observations on chemical excitants of cutaneous pain in man. J Physiol 1953, 120: 326–351.
Armstrong D, Keele CA, Jepson JB, Stewart JW. Development of pain producing substance in human plasma. Nature 1954, 174: 791–792.
Armstrong D, Jepson JB, Keele CA, Stewart JW. Activation by glass of pharmacologically active agents in blood of various species. J Physiol 1955, 129: 80–81.
Armstrong D, Jepson JB, Keele CA, Stewart JW. Pain-producing substance in human inflammatory exudates and plasma. J Physiol 1957, 135: 350–370.
Keele CA. Chemical causes of pain & itch. Proc R Soc Med 1957, 50: 477–484.
Keele CA. The chemistry of pain production. Proc R Soc Med 1967, 60: 419–422.
Bleehen T, Hobbiger F, Keele CA. Identification of algogenic substances in human erythrocytes. J Physiol 1976, 262: 131–149.
Lariviere WR, Melzack R. The bee venom test: a new tonic-pain test. Pain 1996, 66: 271–277.
Chen J, Luo C, Li HL, Chen HS. Primary hyperalgesia to mechanical and heat stimuli following subcutaneous bee venom injection into the plantar surface of hindpaw in the conscious rat: A comparative study with the formalin test. Pain 1999, 83: 67–76.
Chen HS, Chen J. Secondary heat, but not mechanical, hyperalgesia induced by subcutaneous injection of bee venom in the conscious rat: effect of systemic MK-801, a non-competitive NMDA receptor antagonist. Eur J Pain 2000, 4: 389–401.
Chen J, Chen HS. Pivotal role of capsaicin-sensitive primary afferents in development of both heat and mechanical hyperalgesia induced by intraplantar bee venom injection. Pain 2001, 91: 367–376.
Wheeler-Aceto H, Porreca F, Cowan A. The rat paw formalin test: comparison of noxious agents. Pain 1990, 40: 229–238.
Hong Y, Abbott FV. Behavioural effects of intraplantar injection of inflammatory mediators in the rat. Neuroscience 1994, 63: 827–836.
Cooper B, Bomalaski JS. Activation of mechanonociceptors by pro-inflammatory peptides melittin and PLAP peptide. Exp Brain Res 1994, 100: 18–28.
Chen J, Luo C, Li HL. The contribution of spinal neuronal changes to development of prolonged, tonic nociceptive responses of the cat induced by subcutaneous bee venom injection. Eur J Pain 1998, 2: 359–376.
You HJ, Chen J. Differential effects of subcutaneous injection of formalin and bee venom on responses of wide-dynamic-range neurons in spinal dorsal horn of the rat. Eur J Pain 1999, 3: 177–180.
Li MM, Yu YQ, Fu H, Xie F, Xu LX, Chen J. Extracellular signal-regulated kinases mediate melittin-induced hypersensitivity of spinal neurons to chemical and thermal but not mechanical stimuli. Brain Res Bull 2008, 77: 227–232.
Yu YQ, Chen J. Activation of spinal extracellular signaling-regulated kinases by intraplantar melittin injection. Neurosci Lett 2005, 381:194–198.
Yu YQ, Zhao F, Chen J. Activation of ERK1/2 in the primary injury site is required to maintain melittin-enhanced wind-up of rat spinal wide-dynamic-range neurons. Neurosci Lett 2009, 459: 137–141.
Lu ZM, Xie F, Fu H, Liu MG, Cao FL, Hao J, Chen J. Roles of peripheral P2X and P2Y receptors in the development of melittin-induced nociception and hypersensitivity. Neurochem Res 2008, 33: 2085–2091.
Ding J, Xiao Y, Lu D, Du YR, Cui XY, Chen J. Effects of SKF-96365, a TRPC inhibitor, on melittin-induced inward current and intracellular Ca2+ rise in primary sensory cells. Neurosci Bull 2011, 27: 135–142.
Hao J, Liu MG, Yu YQ, Cao FL, Li Z, Lu ZM, Chen J. Roles of peripheral mitogen-activated protein kinases in melittin-indced nociception and hyperalgesia. Neuroscience 2008, 152: 1067–1075.
Ding J, Zhang JR, Wang Y, Li CL, Lu D, Guan SM, Chen J. Effects of a non-selective TRPC channel blocker, SKF-96365, on melittin-induced spontaneous persistent nociception and inflammatory pain hypersensitivity. Neurosci Bull 2012, 28: 173–181.
Yu YQ, Zhao ZY, Chen XF, Xie F, Yang Y, Chen J. Activation of tetrodotoxin-resistant sodium channel NaV1.9 in rat primary sensory neurons contributes to melittin-induced pain behavior. NeuroMol Med 2013, 15: 209–217.
Yu YQ, Zhao F, Guan SM, Chen J. Antisense-mediated knockdown of NaV1.8, but not NaV1.9, generates inhibitory effects on complete Freund’s adjuvant-induced inflammatory pain in rat. PLoS One 2011, 6: e19865.
Yu YQ, Chen XF, Yang Y, Yang F, Chen J. Electrophysiological identification of tonic and phasic neurons in sensory dorsal root ganglion and their distinct implications in inflammatory pain. Physiol Res 2014; 63: 793–799.
This review was supported by grants from the National Basic Research Development Program of China (2013CB835100), the National Natural Science Foundation of China (81171049, 31300919, and 31400948), the National Key Technology R&D Program, China (2013BAI04B04), and the Twelfth Five-Year Project of China (AWS12J004).
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Chen, J., Guan, SM., Sun, W. et al. Melittin, the Major Pain-Producing Substance of Bee Venom. Neurosci. Bull. 32, 265–272 (2016). https://doi.org/10.1007/s12264-016-0024-y
- Spinal dorsal horn