Honey bee venom decreases the complications of diabetes by preventing hemoglobin glycation Javad Behroozi a , Adeleh Divsalar a, ⁎, Ali Akbar Saboury b,c a Department of Cell & Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran b Institute of Biochemistry & Biophysics, University of Tehran, Tehran, Iran c Center of Excellence for Biothermodynamics, University of Tehran, Tehran, Iran abstract article info Article history: Received 14 May 2014 Received in revised form 16 September 2014 Accepted 17 September 2014 Available online 20 September 2014 Keywords: Bee venom Diabetes Hemoglobin Glycation Honey bee venom (BV) and its constituents have been reported to contain a wide variety of pharmaceutical prop- erties such as analgesic, anti-inflammatory, anti-nociceptive, and anticancer effects. Hyperglycemia in diabetes leads to increased protein glycation resulting in structural and functional alteration in proteins. Here, we inves- tigated the effect of BV on the glycation of human hemoglobin. Hemoglobin was incubated with glucose in the presence or absence of BV for 5 weeks. The glycation extent of hemoglobin was examined by UV–visible, Circular Dichroism (CD) and fluorometry methods. Results of the present study showed that BV prevents glycation- induced increasing in β-sheet structure, decreasing in free amino groups, altering in the secondary structure and heme degradation in the hemoglobin. These results imply that BV has a significant antiglycation effect, which can restrain glycation-induced alteration in the secondary structure and function of hemoglobin. Hence, BV has the potential to be used as a natural drug to prevent diabetes complications. © 2014 Elsevier B.V. All rights reserved. 1. Introduction Honey bee (Apis mellifera L.) venom, which is stored by honey bees within their venom sacs for self-defense against a broad diversity of predators, has traditionally been used to treat a variety of diseases and conditions, such as arthritis, rheumatism, back pain, cancerous tumors, and skin disease [1–3]. Bee venom (BV) contains various peptides including mellitin, apamin, adolapin and mast cell degranulation peptide, which have a wide variety of pharmaceutical properties. It also contains enzymes (e.g. phospholipase A2) and non-peptide components (e.g. histamine, lipids and carbohydrates) [4–6]. Two major ingredients of BV are phos- pholipase A and melittin [7]. Melittin is a small protein containing 26 amino acid residues with a molecular weight of 2850 Da and is the prin- cipal toxin in BV [8]. Melittin has been reported to contain pro- inflammatory [9], anti-inflammatory [10], anti-nociceptive [11], and an- ticancer effects [12]. BV exhibits pharmacological actions such as analgesic, anti- arthritic, and anti-inflammatory effects attributable to bioactive compounds [13,14]. It has been used for the treatment of inflammatory diseases such as rheumatoid arthritis and relief of pain in oriental med- icine [15,16]. Besides, previous studies have reported that BV induces apoptotic death in mouse melanoma cells [1]; however, the underlying mechanisms are not clear. BV therapy is done by various methods, such as Apitherapy (using live honey bee stings) [17], apipuncture (bee venom acupuncture) and direct injection of BV [18]. Hyperglycemia, a hallmark of diabetes mellitus (DM), leads to in- creased protein glycation resulting in structural and functional alter- ation in proteins [19]. Diabetic patients are susceptible to long-term complications of diabetes, such as retinopathy [20], cataract [21], ath- erosclerosis [22,23], neuropathy [24], and nephropathy [25] and incom- plete and prolonged wound healing [26]. Protein glycation is the most important factor in the development of these complications [27], and these complications of diabetes as a later result of disordered glu- cose metabolism, are the main reason of morbidity and mortality for patients [28]. Glycation is a non-enzymatic reaction between sugars and a free amino group of proteins resulting in advanced glycation end-products (AGEs) [29]. Protein glycation and AGEs are accompanied by increased free radical activity that leads to the biomolecular damage in diabetes [19]. AGEs generate oxygen free radicals that may potentiate the devel- opment of atherosclerosis [30]. Moreover, AGEs can produce oxygen free radicals through an indirect process, by inducing the release of cy- tokines through interaction of AGEs with their cellular receptors [31]. Because of widespread occurrence of AGEs and the oxidative stress derived from them in a variety of diseases and diabetes complications, it has a great deal of interest to identify and develop AGE inhibitor that can suppress AGE formation [32]. Numerous AGE inhibitors have been developed, such as amino guanidine the most well-known AGE in- hibitor. In animal models, amino guanidine ameliorated diabetic Journal of Molecular Liquids 199 (2014) 371–375 ⁎ Corresponding author. E-mail address: divsalar@khu.ac.ir (A. Divsalar). http://dx.doi.org/10.1016/j.molliq.2014.09.034 0167-7322/© 2014 Elsevier B.V. All rights reserved. Contents lists available at ScienceDirect Journal of Molecular Liquids journal homepage: www.elsevier.com/locate/molliq