Effects of cosmetics containing purified honeybee(Apis mellifera L.) venom on acne vulgaris

Acne vulgaris is a chronic dermatologic problem with multiple factors involved in its pathogenesis. Alternative solutions to acne treatment were instigated by antibiotic resistance despite of its extensive use. Purified bee venom （PBV） has been proposed as a promising candidate for that purpose. The present study was designed to confirm the antibacterial effect of PBV and access the efficacy of cosmetics containing PBV in subjects with acne vulgaris. METHODS： The skin bacterium Propionibacterium acnes was incubated with PBV at various concentrations and bacterial growth was evaluated using the colony forming unit （CFU） assay. The mechanism of PBV employed in killing P. acnes was examined by scanning electron microscopy （SEM） and transmission electron microscopy （TEM）. In addition, a total of 12 subjects were randomized in a double-blind, controlled trial to receive either cosmetics containing PBV or cosmetics without PBV for two weeks. Evaluations included lesion counts and skin microorganism. RESULTS： PBV exhibited antimicrobial activity in a concentration-dependent manner, reducing the number of P. acnes CFU by approximately 6 logs at a concentration of 0.5 mg. When PBV concentration was higher than 1.0 mg, no P. acnes colonies were spotted on an agar. TEM and SEM of untreated P. acnes illustrated the normal pleomorphic structure, whereas the PBV- treated bacterium lost the integrity of surface architecture. Significant difference （P=0.027） in the grading levels based on numbers of lesion counts for inflammatory and noninflammatory was observed in favour of the PBV group compared with the control group. In terms of average decrement of skin microorganism, subjects receiving cosmetics containing PBV experienced a significant 57.5% decrease of adenosine triphosphate levels, whereas participants receiving cosmetics without PBV experienced a nonsignificant decrease of 4.7%. CONCLUSION： These results show that the in vitro actions of antimicrobial activity of PBV were translated in vivo. Cosmetics containing PBV provided a certain degree of efficacy in terms of lesion counts and skin microorganism concentration compared with cosmetics without PBV in subjects with acne vulgaris. PBV may be a good candidate compound for developing therapeutic drua for the treatment of acne vulaaris.

Anti-arthritic effects of microneedling with bee venom gel

Objective:To combine with transdermal drug delivery using microneedle to simulate the bee venom therapy to evaluate the permeation of bee venom gel.Methods:In this study,the sodium urate and LPS were used on rats and mice to construct the model.Bee venom gelemicroneedle combination effect on the model is to determine the role of microneedle gel permeation by observing inflammation factors.Results:Compared with the model group,the bee venom gelemicroneedle combination group can reduce the level of serum nitric oxide of the acute gouty inflammation model caused by sodium urate,and on LPS induced mouse model of acute inflammation effect and the micro.Conclusions:Bee venom can significantly suppress the occurrence of gouty arthritis inflammation in rats and mice LPS inflammatory reaction.Choose the 750 mm microneedle with 10N force on skin about 3 minutes,bee venom can play the optimal role,and the anti-inflammatory effect is obvious.Microneedles can promote the percutaneous absorption of the active macromolecules bee venom gel.

Inhibitory effects of microinjection of morphine into thalamic nucleus submedius on ipsilateral paw bee venom-induced inflammatory pain in the rat

Objective To examine whether microinjection of morphine into the rat thalamic nucleus submedius (Sm) could depress the bee venom (BV)-induced nociceptive behaviours. Methods In inflammatory pain model induced by BV subcutaneous injection into rat unilateral hind paw,the inhibitory effects of morphine microinjection into thalamic nucleus submedius (Sm) on the spontaneous nociceptive behavior,heat hyperalgesia and tactile allodynia,and the influence of naloxone on the morphine effects were observed in the rat. Results A single dose of morphine (5.0 μg,0.5 μL) applied into the Sm ipsilateral to the BV injected paw significantly depressed the spontaneous paw flinching response. Morphine also significantly increased the heat paw withdrawal latencies in the bilateral hind paw and the tactile paw withdrawal threshold in the ipsilateral hind paw 2 hours after BV injection. All these depressive effects could be effectively antagonized by pre-treatment with the opioid receptor antagonist naloxone (1.0 μg,0.5 μL) in the Sm 5min prior to morphine administration. Naloxone alone injected to the Sm had no effect on the BV-induced nociceptive behavior. Conclusion These results suggest that Sm is involved in opioid receptor-mediated anti-nociception in the rat with the BV-induced inflammatory pain. Together with results from previous studies,it is likely that this effect is produced by activation of the Sm-ventrolateral orbital cortex-periaqueductal gray pathway,leading to activation of the brainstem descending inhibitory system and depression of the nociceptive inputs at the spinal cord level.

In Vitro Study on the Effect of Bee Venom on Some Cell Lines and Lumpy Skin Disease Virus

Bee venom （BV） was used from long time ago in the medical field as treatment of chronic joint affections. In the recent decades, the screening process of new sources of antimicrobials discovers its high advantageous characteristics for combating various types of microbes, as well as trials to discover its anti-cancer medicinal fields. Lumpy skin disease virus （LSDV） causes disease in cattle of economic importance, and this work aimed to find treatment as well as alternative inactivant for LSDV. The use of bee venom as antiviral was experimented in this work and exhibited satisfied inhibitory effects on LSDV, meanwhile, the antigenic properties was still intact. The viability of virus was tested in tissue culture cells lines and in embryonated chicken eggs. According to doses and time of exposure, the cell lines of Hep-2 （human larynx carcinoma） and MCF7 （breast carcinoma cell line） were treated with different concentrations of BV and examined after 24 h post-inoculation. The Hep-2 and MCF7 cell lines were treated with various concentrations of BV in descending doses as follow： 25, 20, 15, 10, 5 and 0.5 ug/mL of BV. Then bee venom pathological effects on Hep-2 cells and MCF7 cells were observed, such as apoptosis, retarded growths and cytolysis. The results indicate the possibilities of using bee venom as anti-neoplastic and antiviral.

Effect of natural extract eye drops in dry eye disease rats

AIM:To investigate the therapeutic effect of natural extract eye drops containing bee venom,musk,and deer antlers in dry eye disease(DED)animal models.METHODS:Scopolamine-injected DED rats and lacrimal gland-excised rats were allocated into control,saline,and natural extract groups respectively and a normal group(lacrimal gland excision was not performed)in lacrimal gland-excised rats.After eye drop instillation 4 times a day for 5 d,corneal fluorescein staining(CFS)scores,tear MUC5AC levels,and tear lactic dehydrogenase(LDH)levels were measured.RESULTS:In scopolamine-injected rats,the natural extract-treated group had significantly lower CFS scores(1.7±0.5,4.7±1.4,3.8±1.9,P=0.006)and tear LDH levels(0.10±0.01,0.19±0.01,0.16±0.08 OD,P=0.014)but higher tear MUC5AC levels(12.9±3.7,7.9±2.0,9.7±3.6 ng/m L,P=0.041)compared with the control and saline-treated groups.There were no significant differences between the control and saline-treated groups.In lacrimal gland-excised rats,the natural extract-treated group also had lower CFS scores(4.3±1.2,11.5±2.3,9.0±1.9,P<0.001,P=0.001)and tear LDH levels(0.30±0.08,0.48±0.12,0.39±0.05 OD,P<0.05)but higher tear volume(4.3±0.9,1.9±0.7,2.8±1.1 mm,P=0.005,P=0.124)and tear MUC5 AC levels(8.2±2.0,2.9±1.2,5.4±2.2 ng/m L,P<0.001,P=0.047)compared with the control and saline-treated groups.There were no significant differences in the CFS scores,tear MUC5AC level,and tear LDH level between the normal and natural extracttreated groups.CONCLUSION:The natural extract consisting of bee venom,musk,and deer antlers may have effectiveness in DED treatment by restoring the damaged ocular surface,increasing tear volume,and recovering the tear mucin layer in DED rats.

Effectiveness of bee venom acupuncture in alleviating post-stroke shoulder pain: a systematic review and meta-analysis

BACKGROUND：Shoulder pain is a common complication of stroke.Bee venom acupuncture（BVA）is increasingly used in the treatment of post-stroke shoulder pain.OBJECTIVE：To summarize and evaluate evidence on the effectiveness of BVA in relieving shoulder pain after stroke.SEARCH STRATEGY：Nine databases,namely MEDLINE,EMBASE,the Cochrane Library,the China National Knowledge Infrastructure（CNKI）,the Japan Science and Technology Information Aggregator,Electronic（J-STAGE）,and four Korean medical databases,namely,the National Assembly Library,the Research Information Service System,the National Discovery for Science Leaders,and OASIS,were searched from their inception through August 2014 without language restrictions.INCLUSION CRITERIA：Randomized controlled trials（RCTs）were included if BVA was used at acupoints as the sole treatment,or as an adjunct to other treatments,for shoulder pain after stroke.DATA EXTRACTION AND ANALYSIS：Two review authors independently selected trials for inclusion,assessed methodological quality and extracted data.RESULTS：A total of 138 potentially relevant articles were identifi ed,4 of which were RCTs that met our inclusion criteria.The quality of studies included was generally low,and a preponderance of positive results was demonstrated.All four trials reported favorable effects of BVA on shoulder pain after stroke.Two RCTs assessing the effects of BVA on post-stroke shoulder pain,as opposed to saline injections,were included in the meta-analysis.Pain was signifi cantly lower for BVA than for saline injections（standardized mean difference on 10-cm visual analog scale：1.46 cm,95%CI=0.30–2.62,P=0.02,n=86）CONCLUSION：This review provided evidence suggesting that BVA is effective in relieving shoulder pain after stroke.However,further studies are needed to confi rm the role of BVA in alleviating post-stroke shoulder pain.Future studies should be conducted with large samples and rigorous study designs.

Bee wax coated water-soluble fraction of bee venom improved altered glucose homeostasis in streptozotocin-induced diabetic rats

OBJECTIVE:To evaluate the anti-diabetic efficacy of orally administered bee wax coated water-soluble fraction of bee venom(BWCBVA)drug over orally administered bee wax(BW)and intraperitoneally administered whole bee venom(BV)in streptozotocin(STZ)-induced diabetic rats.METHODS:Diabetes induced by intraperitoneal administration of 60 mg/kg STZ was treated with BWCBVA,BW,and BV for 21 d.The biochemical,protein and histological changes,and physical characteristics of BWCBVA were then analyzed.RESULTS:The BWCBVA group shows significantly decreased blood glucose level as compared to the BW and intraperitoneally administered whole BV treated group.Moreover,BWCBVA significantly normalizes the serum biochemical parameters and increases the body weight.Also,administration of BWCBVA significantly reverses the altered liver expression of phosphatidylinositide 3-kinases-p85 and liver glucokinase.Histological analysis of the pancreas an increase in the islet cell numbers and decrease inβ-cell damage.Co-administering BWCBVA 0.25 mg/kg with nifedipine(6.8 mg/kg)and nicorandil(13.8 mg/kg)to the diabetic rats results in insulin secretion through enhanced calcium ion influx.High performance liquid chromatography and gas chromatography was performed to identify the pharmacologically important compounds present in BWCBVA.CONCLUSION:Our results indicate that BWCBVA,an orally administered colon specific drug delivery system,can be effective in treating diabetes mellitus.

Bee venom phospholipase A2 ameliorates amyloidogenesis and neuroinflammation through inhibition of signal transducer and activator of transcription-3 pathway in Tg2576 mice

Background:Neuroinflammation and accumulation ofβ-amyloid(Aβ)play a significant role in the onset and progression of Alzheimer’s disease(AD).Our previous study demonstrated that signal transducer and activator of transcription-3(STAT3)plays a major role in neuroinflammation and amyloidogenesis.Methods:In the present study,we investigated the inhibitory effect of bee venom phospholipase A2(bvPLA2)on memory deficiency in Tg2576 mice,which demonstrate genetic characteristics of AD and the mechanism of its action at the cellular and animal level.For in vivo study,we examined the effect of bvPLA2 on improving memory by conducting several behavioral tests with the administration of bvPLA2(1 mg/kg)to Tg2576 mice.For in vitro study,we examined the effect of bvPLA2 on amyloidogenesis and neuroinflammation by treating bvPLA2 on LPSactivated BV2 cells.Results:We found that bvPLA2 alleviated memory impairment in Tg2576 mice,as demonstrated in the behavioral tests assessing memory.In the bvPLA2-treated group,Aβ,amyloid precursor protein(APP),and β-secretase 1(BACE1)levels and β-secretase activity were significantly decreased.Expression of pro-inflammatory cytokines and inflammation-related proteins decreased in the brain of bvPLA2-treated group,whereas anti-inflammatory cytokines increased.In addition,bvPLA2 reduced STAT3 phosphorylation in the brains of the bvPLA2-treated group.At the cellular level,bvPLA2 inhibits production of nitric oxide,pro-inflammatory cytokines,and inflammation-related proteins including p-STAT3.Additionally,bvPLA2 inhibits the production of Aβin cultured BV-2 cells.Results from the docking experiment,pull-down assay,and the luciferase assay show that bvPLA2 directly binds STAT3 and,thus,regulates gene expression levels.Moreover,when the STAT3 inhibitor and bvPLA2 were administered together,the anti-amyloidogenic and anti-inflammatory effects were further enhanced than when they were administered alone.Conclusion:These results suggest that bvPLA2 could restore memory by inhibiting the accumulation of Aβ and inflammatory responses via blockage of STAT3 activity.

Effectiveness of bee venom acupuncture for patients suffering from periarthritis humeroscapularis

OBJECTIVE:To evaluate the efficacy of bee venom acupuncture in humeroscapularis(PHS)patients.METHODS:One hundred and twenty patients diagnosed with PHS were assigned into four groups:BV1(0.01 mg/kg),BV2(0.005 mg/kg),BV3(0.0025 mg/kg),and control group(vitamin B1 plus novocain 3%injection)with 15 d of treatment.The outcomes of the study including visual analogue scale(VAS)score andβ-endorphin,inflammatory cytokines including interleukin-10(IL-10),IL-1βand tumor necrosis factorα(TNF-α)and shoulder function score were assessed at baseline,after 10 and 15 d of treatment.RESULTS:All four groups reported statistically significant improvement in VAS score,motion range,and shoulder function score(P<0.01),only the BV3 group showed significant increase of anti-inflammatory(IL-10)and decrease of pro-inflammatory(IL-1β,TNF-α)cytokines after treatment(P<0.05).The BV3 group presented a significant difference between all outcomes compared to the control and other groups.CONCLUSION:BV3 groups showed better recovery including reduced pain,improved motor function and normalized inflammatory cytokines than current therapy used in Vietnam and other groups.

Occupational Asthma Caused by Inhalable Royal Jelly ancl Its Cross-reactivity with Honeybee Venom

Royal jelly is a honeybee nutriment secreted from the glands in the hypopharynx of worker bees essential in the development of queen bees. Ingestion of royal jelly has been reporied to trigger rhinitis, asthma, and anaphylaxis, but occupational asthrna occurring after inhalation of volatile royal jelly is rare.

Bee venom acupuncture reduces neuroinflammation modulating microglia/macrophage phenotype polarization in spinal cord injury compression model

Aim: The present study aimed to examine whether apipuncture (stimulation of acupuncture points with bee venom)at ST36 and GV3 acupoints promotes neuroprotection and reduces neuroinflammation by modulating M1 and M2 phenotype polarization.Methods: Wistar rats were treated with bee venom (BV) (0.08 mg/kg) injection at acupoints ST36 and GV3 [BV (ST36 + GV3)-spinal cord injury (SCI)] or BV injection at non-acupoints [BV (NP)-SCI] or no treatment (CTL-SCI)after SCI by compression. The spinal cord mRNA expression of iNOS, Arg-1 and TGF-β was measured by real time PCR and the levels of IBA-1;BCL-2;NeuN e CNPase was measured by western blotting. Locomotor performance was measured by Basso, Beattie, and Bresnahan (BBB) and grid-walking tests.Results: Apipuncture treatment was able to (1) ameliorate locomotor performance;(2) reduce inflammatory markers (Cox-2 levels) and activation of microglia and macrophages;(3) reduce the polarization of the M1 phenotype marker (iNOS) and increase M2 (Arg-1 and TGF-β) phenotypic markers;(4) promote neuroprotection by reducing the death of neurons and oligodendrocytes;and (5) increase the expression of the anti-apoptotic factor BCL-2.Conclusion: Apipuncture treatment induces locomotor recovery and neuroprotection after the compression model of spinal cord injury. Further, it reduces neuroinflammation by decreasing M1 polarization and increasing M2 phenotype.

Expression of a bee venom phospholipase A_2 from Apis cerana cerana in the baculovirus-insect cell

Bee venom phospholipase A2(BvPLA2) is a lipolytic enzyme that catalyzes the hydrolysis of the sn-2 acyl bond of glycerophospholipids to liberate free fatty acids and lysophospholipids.In this work,a new BvPLA2(AccPLA2) gene from the Chinese honeybee(Apis cerana cerana) venom glands was inserted into bacmid to construct a recombinant transfer vector.Tn-5B-4(Tn) cells were transfected with the recombinant bacmid DNA for expression.Sodium dodecylsulfate-polyacrylamide gel electrophoresis(SDS-PAGE) analysis revealed a double band with molecular weights of 16 and 18 kDa.Products of hexahistidine AccPLA2 fusion protein accumulated up to 5.32% of the total cellular proteins.The AccPLA2 fusion protein was cross reactive with the anti-AmPLA2(BvPLA2 of the European honeybee,Apis mellifera) polyclonal serum.The reaction resulted in a double glycosylation band,which agrees with the band generated by the native AmPLA2 in Western blot analysis.The PLA2 activity of the total extracted cellular protein in the hydrolyzing egg yolk is about 3.16 μmol/(min·mg).In summary,the recombinant AccPLA2 protein,a native BvPLA2-like structure with corresponding biological activities,can be glycosylated in Tn cells.These findings provided fundamental knowledge for potential genetic engineering to produce AccPLA2 in the pharmaceutical industry.

A novel peptide from Apis mellifera and solid-phase synthesis of its analogue

A novel peptide designated secapin-1, was purified and characterized from Apis mellifera. The molecular weight of 25 amino acid peptide secapin-1 was found to be 2821.5625 Da by ESI-FTICR-MS. It showed high identity to secapin. The sequence of secapin-1 was determined to be YIINVPPRCPPGSKFVKNKCRVIVP by automatic Edman degradation. A disulfide bond was formed between Cys9 and Cys20 residues. In addition, an analogue of secapin-1 was synthesized by solid phase peptide synthesis method. The synthesis product was successfully purified and identified to homogeneity by using a combination of SEC, IEC, and RP-HPLC techniques.