Therapeutic potential of infrared-treated bee venom: enhanced multi-faceted bioactivity via compositional modulation.

The new type of coronavirus infection (COVID-19), caused by coronavirus-2 (SARS-CoV-2), has led to a world pandemic due to severe acute respiratory syndrome. In addition to drug and vaccine studies for COVID-19, studies on various foods maintain to increase immunity and alternative treatment, and in this context, bee products are also being researched. Although many studies are showing that bee products have antimicrobial properties and immune-enhancing effects, there are limited studies on the effectiveness of these products against coronavirus. Some peptides in royal jelly are reported to be potent antibacterial and antifungal agents that may be beneficial for avoiding co-infections in COVID-19 patients. Positive results have been found Pollen, a fine and powder-like substance produced by flowering plants and collected by bees, in many studies investigating the effects of pollen on health such as antimicrobial, antiviral and anti-inflammatory. Bee venom; It is a yellowish-colored, bitter-sweet, pungent-smelling substance that is produced in the venom sac of bees, normally in liquid form, but dries up and crystallizes after contact with air. Melittin, the primary component of bee venom having more than 40 biologically and pharmacologically active compounds including phospholipase A2, histamine, epinephrine, free amino acids, peptide and apamin, has been stated to have antitumor, antimicrobial, anti-nociceptive and anti-inflammatory activities. Phospholipase A2 (PLA2) secreted from bee and snake venom is known to have strong anti-HIV activities. Melittin, phosphorylase A2 and hyaluronidan, which are the most significant components of bee venom, constitute 50% of bee venom. Moreover, researches on the relationship between bee venom and COVID-19 are limited. The target of this review is to bring together the studies on the health effects of royal jelly, bee pollen and bee venom, and to contribute to the existing studies.

Bee venom is a medicinal product that is widely used in traditional therapies owing to its excellent anti-inflammatory activity. However, the use of bee venom has shown adverse effects. Therefore, there is a need for research that can remove the cytotoxicity of bee venom and enhance its efficacy. In this study, we hydrolyzed melittin, the main component of bee venom, and removed the other components to eliminate the toxicity of bee venom. To compare the efficacy of bee venom and detoxified bee venom, we examined their antioxidant effects using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. In addition, cytotoxicity was confirmed in MCF 10A and RAW 264.7 cells, using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Detoxified bee venom showed a strong antioxidant activity and decreased a cytotoxicity in MCF 10A and RAW 264.7 cells. The anti-inflammatory activity of detoxified bee venom and bee venom were assessed by comparison of the expression of inflammatory cytokine mRNA and phosphorylation of IκBα in RAW 264.7 cells. Degranulation in RBL-2H3 cells was analyzed through β-hexosaminidase release assay to confirm the allergenic activity of bee venom and detoxified bee venom. Treatment of the detoxified bee venom inhibited inflammatory cytokine mRNA expression, IκBα phosphorylation, and β-hexosaminidase release. Taken together, the results indicated that compared to bee venom, detoxified bee venom exhibited decreased cytotoxicity and allergenicity and increased anti-inflammatory activity. In conclusion, detoxification of bee venom efficiently decreases the adverse effects, making it suitable for medicinal applications.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12010-021-03653-2.

Gentamicin (GM) is a frequently used aminoglycoside for managing serious illnesses; nonetheless, renal complications limit its use. Bee venom (BV) is a biological toxin that exhibits anti-inflammatory and antioxidant activities. This study was designed to explore the mitigating effect of BV remediation on GM induced renal injury. Twenty male rats were divided into four groups (five rats each), namely, control (saline subcutaneously); BV group (1 mg/kg S/C twice weekly for 1 month); GM group (100 mg/kg i. p. for 1 week); and GM-BV group (the same aforementioned dosages of GM and BV, with GM administered in the last week for 4 weeks). BV mitigated the GM-inflicted kidney damage, as evidenced by a substantial improvement in the renal function and oxidative state. In addition, a downregulation in the expression of inflammatory biomarkers (Casp-1, IL-6, TNF-α, and NF-κB/P65/P50) and an upregulation of oxidative stress marker expression (NRF2) were noticed. BV upregulated the expression of aquaporins (AQPs) and renal water channel proteins (AQP1 and AQP2), which are useful for the early detection of renal injury. Additionally, BV exposure exerted a mitigating effect on the apoptotic cascade, as evidenced by the downregulation of cleaved Caspase-3 (Casp-3) and cytochrome c (Cyto c). BV administration also led to an improvement in RBC, WBC, and platelet counts, along with enhanced Hb levels. Interestingly, BV could protect against GM triggered nephrotoxicity.

Background/Objectives: Bee venom (BV), as a natural product, is one of the foundations of the pharmaceutical industry, through which many diseases, including serious ones, can be effectively treated. The BV nanofilm is an effective antidote delivered into the human body to target the affected area and address the issue without major side effects. In this study, we investigated the intriguing therapeutic effects of apitoxin (bee venom) used in isolation, combined with the powerful properties of zinc oxide nanoparticles. Methods and Results: BV nanofilm was evaluated using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD). The BV extract was analyzed using mass spectrometry (MS), which identified 84 active components present at varying concentrations. BV was treated with both polyvinyl alcohol (PVA) and zinc oxide nanoparticles (ZNPs) to increase the intensity of OH and CH2 groups and to enhance the dispersion of C=O. BV has demonstrated anti-type 2 diabetes activity by inhibiting α-amylase and α-glucosidase, which are starch-degrading enzymes. The nanofilm is an active mixture of BV, PVA, and ZNPs, which exhibited the highest antidiabetic activity with IC50 values of 30.33 μg/mL and 5.55 μg/mL for the inhibition of α-amylase and α-glucosidase, compared to IC50 of 51.69 µg/mL and IC50 of 7.30 µg/mL for BV, respectively. The nanofilm also showed higher anti-inflammatory activity by inhibiting red blood cell (RBC) hemolysis, with an IC50 of 16.99 μg/mL in comparison to IC50 of 72.99 µg/mL for BV alone. The nanofilm demonstrated broad-spectrum antimicrobial activity, effectively targeting both Gram-positive (Staphylococcus aureus ATCC 6538 and Bacillus subtilis ATCC 6633) and Gram-negative bacteria (Salmonella typhi ATCC 6539, Escherichia coli ATCC 8739). Furthermore, increased antioxidant activity was recorded by inhibiting the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging effect with an IC50 of 4.26 μg/mL and 19.43 μg/mL for nanofilm and BV, respectively. BV was found to be more toxic to liver tissue (HepG2 cell line) than nanofilm, with IC50 values of 18.5 ± 0.08 μg/mL and 52.27 ± 0.7 μg/mL, respectively. The BV extract displayed higher toxicity to liver tissue (2.3%) with 97.7% viability at 250 μg/mL, compared to nanofilm, which showed 0.09% toxicity and 99.9% viability at the same concentration. Conclusions: the BV nanofilm emerges as a promising alternative medicine, offering an innovative solution for treating various diseases through its high concentration of therapeutically active compounds and effortless targeting delivery.

Anti-cancer effect of bee venom toxin and melittin in ovarian cancer cells through induction of death receptors and inhibition of JAK2/STAT3 pathway

Formaldehyde exposure is associated with inflammation and oxidative stress, leading to systemic toxicity. Natural products such as bee venom have been proposed as alternative therapeutic agents due to their anti-inflammatory and antioxidant properties. This study investigated the potential of Algerian Apis mellifera intermissa bee venom to attenuate formaldehyde-induced acute inflammation and oxidative stress in male rats. Inflammation was induced by subplantar injection of 0.1 mL of a 2.5% formaldehyde solution into the hind paw. One hour later, rats received either subcutaneous bee venom (0.76 mg/kg) or oral diclofenac sodium (10 mg/kg) as a reference drug. Paw edema was quantified using ImageJ software. Blood samples were analyzed for erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), white blood cell count (WBC), platelet count (PLT), and albumin levels. Oxidative stress biomarkers including malondialdehyde (MDA), catalase (CAT), glutathione (GSH), and glutathione S-transferase (GST) were assessed in erythrocytes. Formaldehyde exposure produced sustained paw edema, impaired body weight gain, reduced food and water intake, systemic inflammatory alterations, and pronounced oxidative stress characterized by increased lipid peroxidation and depletion of erythrocyte antioxidant defenses. Bee venom treatment significantly attenuated paw swelling, improved systemic inflammatory alterations, and partially restored redox balance, with effects comparable to those of diclofenac. The overall protective effects of bee venom were comparable to those of diclofenac. These findings demonstrate that A. mellifera intermissa venom exerts significant anti-inflammatory and antioxidant actions and may represent a promising natural therapeutic candidate for inflammation associated with oxidative stress.

Bee venom has long been used as a traditional folk medicine in Korea. It has been reportedly used for the treatment of arthritis, cancer, and inflammation. Although its anti-inflammatory activity in lipopolysaccharide- (LPS-) stimulated inflammatory cells has been reported, the exact mechanism of its anti-inflammatory action has not been fully elucidated. Therefore, the aim of this study was to investigate the anti-inflammatory mechanism of bee venom in BV2 microglial cells. We first investigated whether NO production in LPS-activated BV2 cells was inhibited by bee venom, and further iNOS mRNA and protein expressions were determined. The mRNA and protein levels of proinflammatory cytokines were examined using semiquantitative RT-PCR and immunoblotting, respectively. Moreover, modulation of the transcription factor NF-κB by bee venom was also investigated using a luciferase assay. LPS-induced NO production in BV2 microglial cells was significantly inhibited in a concentration-dependent manner upon pretreatment with bee venom. Bee venom markedly reduced the mRNA expression of COX-2, TNF-α, IL-1β, and IL-6 and suppressed LPS-induced activation of MyD88 and IRAK1 and phosphorylation of TAK1. Moreover, NF-κB translocation by IKKα/β phosphorylation and subsequent IκB-α degradation were also attenuated. Thus, collectively, these results indicate that bee venom exerts its anti-inflammatory activity via the IRAK1/TAK1/NF-κB signaling pathway.

Oxidative damage and proinflammatory cytokines are involved in exhaustive exercise-induced fatigue. This study aimed to investigate the effects of bee venom, a natural toxin, on fatigue and tissue damage in rats that underwent forced swimming exercise. Rats were divided into four groups: control, swimming exercise (SE), bee venom (BV) and swimming exercise + bee venom (SE + BV). SE and SE + BV groups were subjected to forced swimming (load of 7% body weight) for 5days. BV and SE + BV groups were injected with 1mg/kg BV subcutaneously. Swimming time, blood lactate and TNF-α levels, MDA and GSH levels in liver and gastrocnemius muscle were evaluated. Swimming time was shorter in SE + BV group than SE group. There was no difference in lactate levels between SE and SE + BV groups. MDA and GSH levels were increased in SE, BV and SE + BV groups. TNF-α levels were increased in BV group compared to control and SE groups. Our study demonstrated that BV administration before exhaustive exercise in rats did not provide anti-fatigue effect. Additionally, BV did not show anti-inflammatory activity and had different effects on antioxidant capacity at tissue level. Further research might explore the effects of different doses and durations of BV on exhaustive exercise.

Bee venom (BV), or apitoxin, is a complex substance produced by a gland in the abdominal cavity of bees. The main component of BV is melittin, which is a largely studied substance due to its biological properties. To date, the most well-known bee venom and melittin are derived from domesticated honey bees, while venom and melittin derived from wild honey bees have been under-investigated. Hence, this study primarily reports the antimicrobial activities of bee venom and synthetic melittin derived from four different honey bee species (Apis mellifera, A. cerana, A. dorsata, and A. florea) in Thailand. All the bee venom extracts and melittins showed more robust antibacterial activities against Gram-positive (Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus, S. aureus MRSA, and S. epidermidis) than Gram-negative bacteria (Escherichia coli, Klebsiella pneuminiae, and Salmonella typhimurium) or a fungus (Candida albicans), while the synthetic melittins also have antimicrobial activity at higher concentrations than the bee venom extract. Furthermore, the A. cerana venom extract showed the highest activity against the tested bacteria, followed by A. mellifera, A. florea, and A. dorsata. Therefore, A. cerana venom may be further developed for use in medical applications as a potential alternative agent against Gram-positive bacteria and antibiotic-resistant bacteria.

Ethanolic extracts of seasonally collected natural bee products (honey, propolis, royal jelly (RJ), and bee venom (BV)) were tested for their potential as antimicrobial agents against antibiotic-resistant bacteria and fungi. These extracts exhibited various inhibitory effects on antibiotic-resistant bacteria (Streptococcus pneumoniae, Staphylococcus aureus, MRSA, Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus vulgaris, and Haemophilus influenzae) and fungi (Aspergillus brasiliensis and Candida albicans), with the exception of S. pneumonia, which was not inhibited by honey and RJ extracts, and P. aeruginosa, which was not inhibited by RJ extracts. Interestingly, extracts of BV and its major content, melittin (MEL), displayed a wide spectrum of antimicrobial activity against all tested bacteria and fungi. This is the first study to show that propolis extract has bactericidal activity against S. pneumoniae and that BV extract and MEL have antibacterial activity against P. vulgaris, H. influenzae, and H. influenzae type b. Extracts of bee products collected in the spring generally exhibited the most significant antibacterial and antifungal activities. Based on total phenolic content (TPC) and total flavonoid content (TFC), it was found that spring samples of propolis, RJ, and honey, in that order, were the richest. Also, LC-MS-MS analysis of MEL content in BV demonstrated that it was the highest in spring sample. In terms of MIC and MBC values, Gram-positive bacteria were the most susceptible to bee products. First and foremost, the antimicrobial activity of bee products was ranked in descending order based on MIC values: BV, MEL, propolis, RJ, and honey.

For last couple of decades, natural products have served us well in combating different types of cancer. The main sources of these useful compounds are from both animal and plant origin. Here we will present anticancer ability of bee venom (BV) and apigenin (API) towards different types of cancer cells in vitro. BV from honey bees is a complex mixture of a variety of different active peptides while API is a natural flavonoid found in several dietary plant foods. Anticancer effect of whole BV was tested in human cervical carcinoma HeLa cells and their drug-resistant HeLa CK subline while anticancer effect of API was tested in human breast cancer MCF-7 and MDA MB-231 cells. Cytotoxicity of both compounds towards cancer cells was evaluated by MTT assay whereas type of cell death was analysed by differential staining using acridine orange/ethidium bromide and was further verified by Western blot analysis. BV displayed dose-dependent cytotoxicity against both cell lines tested with drug-resistant HeLa CK cells being more sensitive to BV than their parental cell lines. Similarly, API inhibited the growth of both cell lines in a dose-dependent manner with MCF-7 cells being more sensitive. Treatment with BV induced a necrotic type of cell death, as shown by characteristic morphological features, fast staining with ethidium bromide and a lack of cleavage of apoptotic marker poly (ADP-ribose) polymerase (PARP) on Western blot. On the contrary, cell treated with API showed apoptosis as a dominant type of cell death in both cell lines which was further verified by Western blot analysis detecting cleaved PARP. In view of accumulating evidence on anti-proliferative and pro-cell death activity, both tested compounds could be used in the development of future anticancer drugs. Undoubtedly, therapeutic applications of BV and API are promising, however further in vitro and in vivo studies are warranted to resolve precise mechanisms responsible for their anticancer properties

This study aims to assess the antirheumatic activity of bee venom (BV) and/or hesperidin as natural products in complete Freund's adjuvant (CFA)-induced arthritis in male Wistar rats. Rheumatoid arthritis was induced in 30 male Wistar rats (weight 130 g to 150 g; age 10 to 12 weeks) by subcutaneous injection of CFA into the right hind paw of the rats. The rats were divided into five groups of six rats in each and administered the following regimens for 21 days: Normal group (given the equivalent volume of saline and carboxymethylcellulose), arthritic group (given the equivalent volume of saline and carboxymethylcellulose), arthritic group treated with BV (treated with BV along with carboxymethylcellulose), arthritic group treated with hesperidin (treated with hesperidin along with saline), and arthritic group treated with BV and hesperidin (treated with BV and hesperidin concurrently). Bee venom and/or hesperidin successfully reversed the CFA-arthritis-induced increases in right hind leg paw swelling, leukocytes' count, liver lipid peroxidation, serum inflammatory cytokine interleukin (IL-2 and IL-12) levels and spleen tumor necrosis factor-alpha messenger ribonucleic acid expression. Moreover, the CFA-induced down-regulation in serum IL-10 level and spleen IL-4 messenger ribonucleic acid expression as well as the deterioration in the antioxidant defense system were significantly improved as a result of BV and hesperidin administration. Both treatments also markedly counteracted the severe inflammatory changes and leukocytic infiltration in the periarticular tissue of the ankle joints. In addition, BV and hesperidin obviously amended the lymphoid hyperplasia in white pulps of spleen as well as the widening of the medulla and mononuclear cell infiltration found in thymus. Bee venom and hesperidin administration produced their ameliorative effects on rheumatoid arthritis via their antioxidant, antiinflammatory and immunomodulatory potentials. BV plus hesperidin particularly seemed to be the most potent in improving rheumatoid arthritis in Wistar rats.

High levels of blood glucose and lipids are well-known risk factors for heart diseases. Bee venom is a natural product that has a potent hypoglycemic, hypolipidemic, anti-inflammatory, and antioxidant effects. The current study aimed to determine the bee venom effects on cardiac dysfunction compared to combined therapy of metformin and atorvastatin in diabetic hyperlipidemic rats. The median lethal dose of bee venom was estimated, and then 50 adult male albino rats were categorized into five groups. One group was fed a standard diet and served as a negative control, while the other groups were given nicotinamide and streptozotocin injections to induce type 2 diabetes. After confirming diabetes, the rats were fed a high-fat diet for four weeks. The four groups were divided as follows: one group served as a positive control, whereas the other three groups were treated with bee venom (0.5 mg/kg), bee venom (1.23 mg/kg), and combined therapy of metformin (60 mg/kg) and atorvastatin (10 mg/kg), respectively, for four weeks. Upon termination of the experiment, blood samples and heart tissue were obtained. Administration of bee venom using both doses (0.5 and 1.23 mg/kg) and combined therapy of metformin and atorvastatin revealed a significant decrease in the concentrations of glucose, total cholesterol, triacylglycerol, low-density lipoprotein cholesterol, very low-density lipoprotein cholesterol, troponin I, creatine kinase, and lactate dehydrogenase activities. Moreover, a significant decrease had been detedcted in malondialdehyde, nuclear factor-kappa-β levels, and relative mRNA expression of vascular cell adhesion molecule-1 and galectin-3 in heart tissue compared to the positive control (P < 0.0001). Furthermore, there was a significant increase in bodyweight levels of insulin, high-density lipoprotein cholesterol, and total antioxidant capacity in heart tissue compared to the positive control (P < 0.0001). The results indicate that bee venom can ameliorate cardiac dysfunction through attenuating oxidative stress and downregulating the NF-κβ signaling pathway.

Around the globe, human societies have employed insects and the compounds collected from them as a source of therapeutic resources. These creatures have not only been used medically, but also mystically and magically in a variety of civilizations to cure various diseases. For pharmaceutical study, insects seem to be an almost limitless resource. Medicinal potential of insects makes a substantial contribution to the debate over biodiversity preservation.&#x0D; Bee venom treatment is common in conventional medicine to cure ailments including rheumatism, arthritis, discomfort, malignant tumors, and skin. Several peptides with a range of medicinal benefits are present in bee venom including Melittin, apamin, ado lapin, the mast cell degranulating peptide, enzymes (phospolipase-A2) and amines including histamine and adrenaline. Melittin and phospolipase-A2 may be used to treat cancer cells, which can include leukemia and cancer cells of the kidney, liver, prostate, lung, and mammary gland. Bee venom may cause cancer cells to undergo apoptosis, according to a recent study by Moon et al. In rheumatoid synovial cells, bee venom promotes apoptosis by decreasing the expression of BCL2 and increasing the expression of BAX and caspase-3. In synovial fibroblasts from rheumatoid arthritis patients, bee venom causes apoptosis by activating caspase-3 [1].&#x0D; Human immunodeficiency virus can be eliminated by a toxin present in bee venom (HIV). Melittin, which surrounds the HIV virus among other viruses, is present in bee venom. Nanoparticles in this melittin are abundant and target a crucial component of the virus' structure. For use in upcoming clinical studies, nanoparticles are simple to produce in large numbers [2]. Maggot treatment is a kind of biotherapy that includes injecting live, sterilized maggots (fly larvae) into the nonhealing skin and soft tissue wounds of a person or an animal in order to debride the wound of necrotic (dead) tissue and disinfect it. Maggot treatment has been shown to aid in wound healing. The Pseudomyrmex sp. often known as the samsum ant, is a species of South American tree ant. Its venom has a wide range of therapeutic benefits, including the treatment of hepatitis and the protection of the liver [3].&#x0D; The utilization of insects as a natural product has the potential to provide a treatment that is effective in both treating and preventing illnesses. Development of insects as significant new alternative medicines has advanced significantly in recent years. Since insects are very diverse and have long used a wide variety of natural chemicals to adapt to environmental changes, this is an intriguing and quickly growing new field to study in medicine

Bee venom is used as a traditional medicine for treatment of arthritis. The anti-inflammatory activity of the n-hexane, ethyl acetate, and aqueous partitions from bee venom (Apis mellifera) was studied using cyclooxygenase (COX) activity and pro-inflammatory cytokines (TNF-alpha and IL-1beta) production, in vitro. COX-2 is involved in the production of prostaglandins that mediate pain and support the inflammatory process. The aqueous partition of bee venom showed strong dose-dependent inhibitory effects on COX-2 activity (IC50 = 13.1 microg/mL), but did not inhibit COX-1 activity. The aqueous partition was subfractionated into three parts by molecular weight differences, namely, B-F1 (above 20 KDa), B-F2 (between 10 KDa and 20 KDa) and B-F3 (below 10 KDa). B-F2 and B-F3 strongly inhibited COX-2 activity and COX-2 mRNA expression in a dose-dependent manner, without revealing cytotoxic effects. TNF-alpha and IL-1beta, are potent pro-inflammatory cytokines and are early indicators of the inflammatory process. We also investigated the effects of three subfractions on TNF-alpha and IL-1beta production using ELISA method. All three subfractions, B-F1, B-F2 and B-F3, inhibited TNF-alpha and IL-1beta production. These results suggest the pharmacological activities of bee venom on anti-inflammatory process include the inhibition of COX-2 expression and the blocking of pro-inflammatory cytokines (TNF-alpha, and IL-1beta) production.