Therapeutic potential of royal jelly to control Toxoplasma gondii infection in mice

@#At present, there are several synthetic medications for toxoplasmosis therapy; however, these agents cannot be permanently applied because of adverse side effects or therapeutic failures and drug resistance in parasites. The present experimental investigation was aimed to study the effects of royal jelly (RJ) obtained from Apis mellifera in comparison with atovaquone against Toxoplasma gondii infection in mice. After treatment of infected mice with RJ at the doses of 200, 400, and 600 mg/kg for 14 consecutive days, we evaluated the therapeutic activity of RJ by measuring the mean number and the mean size of T. gondii tissue cysts, oxidant-antioxidant enzymes, pro-inflammatory cytokines, the mRNA expression levels of bradyzoite surface antigen 1 (BAG1), as well as the toxic effect on liver and kidney function. Treatment of the infected mice with RJ significantly (p < 0.001) decreased the mean number and the mean diameter of T. gondii tissue cysts and downregulated BAG1 in a dose-dependent response. After treatment of infected mice with RJ, the level of oxidative stress markers was significantly diminished, but a significant increase (p < 0.05) in the level of antioxidant markers such as glutathione peroxidase (GPx) and superoxide dismutase (SOD) enzymes was observed. Treatment of the infected mice with RJ significantly enhanced the level of pro-inflammatory cytokines IFN-γ and IL-1β, whereas it caused no substantial change in the serum levels of liver and kidney enzymes. The findings of this in vivo study revealed the favorable therapeutic effect of RJ on latent T. gondii infection in mice. It was found that RJ considerably inhibited the infection by decreasing the number and size of tissue cysts, reducing oxidative stress, and boosting the level of pro-inflammatory cytokines, but had no significant toxic impact on the function of vital organs such as liver and kidney. However, additional surveys are required to confirm these findings and clarify the exact mechanisms and their efficiency in clinical subjects.

Spores of Paenibacillus larvae, Ascosphaera apis, Nosema ceranae and Nosema apis in bee products supervised by the Brazilian Federal Inspection Service

ABSTRACT Due to their ecological and economic importance, honey bees have attracted much scientific attention, which has intensified due to the recent population decline of these insects in the several parts of the world. Among the factors related to these patterns, infection by pathogens are the most relevant, mainly because of the easy dissemination of these microorganisms. Although no zoonotic diseases are associated with these insects, the presence of infectious agents in bee products should still be considered because they play a role as disease dispersers, increasing the risk to animal health. Because of the possibility of dispersion of pathogens via bee products, this work aimed to identify the presence of spores of the pathogens Paenibacillus larvae, Ascosphaera apis and Nosema spp. in samples of honey, pollen and royal jelly that are registered with Brazil's Federal Inspection Service (S.I.F.) and commercially available in the state of São Paulo. Of the 41 samples of bee products analyzed, only one showed no contamination by any of these pathogens. N. ceranae and P. larvae had the highest prevalence considering all the samples analyzed (present in 87.80% and 85.37% of the total, respectively), with N. apis present in 26.83% and A. apis present in 73.17% of the samples. These results provide support for the formulation of government regulations for sanitary control of exotic diseases by preventing dispersion of pathogens, including through illegal importation, since local and international trade and the transfer of colonies between regions play important roles in the dispersion of these microorganisms.

Anti-senescence effect and molecular mechanism of the major royal jelly proteins on human embryonic lung fibroblast (HFL-I) cell line / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Royal jelly (RJ) from honeybee has been widely used as a health promotion supplement. The major royal jelly proteins (MRJPs) have been identified as the functional component of RJ. However, the question of whether MRJPs have anti-senescence activity for human cells remains. Human embryonic lung fibroblast (HFL-I) cells were cultured in media containing no MRJPs (A), MRJPs at 0.1 mg/ml (B), 0.2 mg/ml (C), or 0.3 mg/ml (D), or bovine serum albumin (BSA) at 0.2 mg/ml (E). The mean population doubling levels of cells in media B, C, D, and E were increased by 12.4%, 31.2%, 24.0%, and 10.4%, respectively, compared with that in medium A. The cells in medium C also exhibited the highest relative proliferation activity, the lowest senescence, and the longest telomeres. Moreover, MRJPs up-regulated the expression of superoxide dismutase-1 (SOD1) and down-regulated the expression of mammalian target of rapamycin (MTOR), catenin beta like-1 (CTNNB1), and tumor protein p53 (TP53). Raman spectra analysis showed that there were two unique bands related to DNA synthesis materials, amide carbonyl group vibrations and aromatic hydrogens. These results suggest that MRJPs possess anti-senescence activity for the HFL-I cell line, and provide new knowledge illustrating the molecular mechanism of MRJPs as anti-senescence factors.

Identification of genes related to high royal jelly production in the honey bee (Apis mellifera) using microarray analysis

Abstract China is the largest royal jelly producer and exporter in the world, and high royal jelly-yielding strains have been bred in the country for approximately three decades. However, information on the molecular mechanism underlying high royal jelly production is scarce. Here, a cDNA microarray was used to screen and identify differentially expressed genes (DEGs) to obtain an overview on the changes in gene expression levels between high and low royal jelly producing bees. We developed a honey bee gene chip that covered 11,689 genes, and this chip was hybridised with cDNA generated from RNA isolated from heads of nursing bees. A total of 369 DEGs were identified between high and low royal jelly producing bees. Amongst these DEGs, 201 (54.47%) genes were up-regulated, whereas 168 (45.53%) were down-regulated in high royal jelly-yielding bees. Gene ontology (GO) analyses showed that they are mainly involved in four key biological processes, and pathway analyses revealed that they belong to a total of 46 biological pathways. These results provide a genetic basis for further studies on the molecular mechanisms involved in high royal jelly production.

Beekeeping and Bee Products; Boon for Human Health and Wealth.

The cross-fertilization by bee increases average agricultural yield by 20 to 25 percent. Its products like honey, pollen, royal jelly, propolis, bee venom have immense medical importance. Honey is useful for healing the wounds, helps to build up hemoglobin, used as laxative blood purifier, preventive against cold, cough, sore throat, eye ailments, burns and gastrointestinal disorder etc. Honey has antibiotic property and is effective in reducing the risk of heart disease, cancer and diabetes. Pollen lowers blood pressure, increases hemoglobin and erythrocyte content, useful in pernicious anemia, sterility, hypertension, in complaints of the nervous and endocrine system. Royal jelly has antimicrobial, anti-inflammatory, anti-aging, vasodilative and hypotensive, antioxidant, antihypercholesterolemic and antitumor property. Royal jelly has a diuretic effect, prevents obesity, builds up resistance to infection, regulates the functioning of the endocrine glands and is good for arteriosclerosis and coronary deficiency. Bee venom acts as antibiotic and useful for lowering of blood pressure, in neural disorders and rheumatoid arthritis and acute rheumatic carditis, treating certain eye diseases, hypertension and gynecological and children’s diseases. Propolis, a resinous substance has pharmacologically active constituents as flavonoids, phenolics and other various aromatic compounds. Propolis has antibacterial, antifungal, antiviral, antioxidant and anti-inflammatory proprieties. It is used to treat mouth and gum disorders, gum decay, resistance to general illness, cure burns and fungal skin complaints. Beeswax is used to prepare polishes, waterproofing, electrical insulation, cosmetics, cold creams etc. It is also useful in engineering, pharmaceutical and confectionary industries.

Reduction Effect of Royal Jelly and Rape Honey Alone and in Combination Against Methicillin-Resistant Staphylococcus aureus (MRSA) Strains

Multidrug resistant and methicilin-resistant Staphylococcus aureus (MRSA) is involved in severe difficult to treat skin and soft tissue infections in humans. In the present study the antibacterial reduction effect of royal jelly (RJ), rape honey (RH), as well as in combination (RJ:RH, 1:100 w/w) against multidrug resistant MRSA strains was evaluated by means of a microbiological method "in vitro". Royal jelly and rape honey mixture possessed a higher antibacterial activity than rape honey. The concentrations of royal jelly (20 and 30% v/v) had a total inhibitory effect against tested MRSA strains. Royal jelly alone and in rape honey mix (RJ:RH, 1:100 w/w) have a potential as alternative therapeutics against MRSA strains, resistant for antibiotics.

Anti-inflammatory effects of royal jelly on ethylene glycol induced renal inflammation in rats

ABSTRACT Objective: In this study, anti-inflammatory effects of Royal Jelly were investigated by inducing renal inflammation in rats with the use of ethylene glycol. For this purpose, the calcium oxalate urolithiasis model was obtained by feeding rats with ethylene glycol in drinking water. Materials and Methods: The rats were divided in five study groups. The 1st group was determined as the control group. The rats in the 2nd group received ethylene glycol (1%) in drinking water. The rats in the 3rd group were daily fed with Royal Jelly by using oral gavage. The 4th group was determined as the preventive group and the rats were fed with ethylene glycol (1%) in drinking water while receiving Royal Jelly via oral gavage. The 5th group was determined as the therapeutic group and received ethylene glycol in drinking water during the first 2 weeks of the study and Royal Jelly via oral gavage during the last 2 weeks of the study. Results: At the end of the study, proinflammatory/anti-inflammatory cytokines, TNF-α, IL-1β and IL-18 levels in blood and renal tissue samples from the rats used in the application were measured. Conclusion: The results have shown that ethylene glycol does induce inflammation and renal damage. This can cause the formation of reactive oxygen species. Royal Jelly is also considered to have anti-inflammatory effects due to its possible antiradical and antioxidative effects. It can have positive effects on both the prevention of urolithiasis and possible inflammation during the existing urolithiasis and support the medical treatment.

The immunomodulatory effects of lyophilized royal jelly on immunoglobulin IgG,IgA,IgM and complement C3,C4 in patients with low immunity / 中国基层医药

Objective To investigate the immunomodulatory effects of lyophilized royal jelly ( LRJ) on immunoglobulin IgG,IgA,IgM and complement C3 and C4 in patients with low immunity ,and to evaluate the clinical efficacy of LRJ.Methods A self-controlled clinical study of immunomodulatory efficacy of LRJ was performed in 32 patients with low immunity.Every patient took orally LRJ,two times a day.The hepatic function AST,ALT,and immunoglobulin IgG,IgA,IgM and complement C3,C4 were assayed after treatment for 28 days,and then the efficacy and safety of LRJ were evaluated.Results After took orally LRJ,the levels of IgG,IgA,and IgM [(10.31 ±2.03)g/L, (1.69 ±0.55)g/L,(1.51 ±0.55)g/L] were significantly higher than those before treatment [(8.52 ±1.94)g/L, (1.38 ±0.57)g/L,(1.36 ±0.55)g/L](t=5.21,2.25,1.56,all P0.05).Conclusion LRJ can increase the levels of immu-noglobulin IgG,IgA and IgM,thus to improve the symptoms of patients with low immunity .

Cytoprotective Potential of Royal Jelly on Human Umbilical Vein Endothelial Cells against Nicotine Toxicity via Catalase.

Aims: To examine the cytoprotective effects and mechanisms of a royal jelly extract in protecting the human umbilical vein endothelial cells (HUVECs) from nicotine toxicity. Study Design: Laboratory experimental tests. Place and Duration of Study: Department of Physiology and Department of Surgery, Faculty of Medicine, Srinakharinwirot University, Bangkok 10110, Thailand, between June 2011 and February 2012. Methodology: Cytotoxic assay of royal jelly to HUVECs was performed by using the 3- (4,5-dimethylthiazol,2-yl)-2,5-diphenyltetrazolium bromide (MTT) reagent. The cytoprotective effect was then investigated by examining the presence of vacuole-like structures in HUVECs exposed to nicotine 5 or 7.5 mM with and without royal jelly. Cells were stained with crystal violet and photographed under phase contrast microscope. mRNA levels of genes involved in intracellular antioxidant system, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione reductase (GSR) were confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Catalase activity was also determined by examining peroxidative function. Results: Vacuole-like structures were found in the cytoplasm of HUVECs exposed to 5 mM nicotine and higher. Royal jelly alone at the concentrations lower than 2 mg/ml did not affect the structure or the survival rate of HUVECs after 1, 4, and 7 days of treatment. For cytoprotective effect, royal jelly 1-4 mg/ml mixed with 5 mM nicotine could obviously decrease the numbers of cells containing vacuole-like structures in the cytoplasm of HUVECs with the dose- and time-dependent fashion. The catalase mRNA levels and catalase activity in HUVECs exposed to 5 mM nicotine decreased significantly, but recovered when the cells were treated with royal jelly. Conclusion: Royal jelly can be safety applied to endothelial cells even at high doses. Royal jelly is able to attenuate the abnormal vacuole-like structures induced in endothelial cell cytoplasm when exposed to nicotine. Further investigation of antioxidant gene expression showed that the mechanism possibly involves a reduction of oxidative stress via an up-regulation of catalase. Besides the supplementary food, royal jelly could be useful for endothelial cell protection from nicotine toxicity found in smoking or nicotine addiction.

Dietary effect of royal jelly supplementation on epidermal levels of hydration, filaggrins, free amino acids and the related enzyme expression in UV irradiated hairless mice

Ultraviolet (UV) irradiation reduces epidermal hydration, which is paralleled by the reduction of natural moisturizing factors (NMFs). Of various NMFs, free amino acids (AAs) are major constituents generated by filaggrin degradation. In this study, we attempted to determine whether dietary supplementation of royal jelly (RJ) in UV-irradiated mice can alters epidermal levels of hydration, filaggrins, and free AAs as well as of peptidylarginine deiminase-3 (PAD3), an enzyme involved in filaggrin degradation processes. Albino hairless mice were fed either a control diet (group UV+: UV irradiated control) or diets with 1% RJ harvested from different areas in Korea (groups RJ1, RJ2, and RJ3) or imported from China (group RJ4) for six weeks in parallel with UV irradiation. A normal control group (group UV-) was fed a control diet without UV irradiation for six weeks. Reduced epidermal levels of hydration, total filaggrins, and PAD3 were observed in group UV+; in group RJ1, these levels were increased to a level similar to that of group UV-. In addition, profilaggrins, two repeat intermediates (2RI), a precursor with two filaggrin repeats, and filaggrin were increased. Although no alteration of AAs was observed in any of the groups, and glutamate and serine, major AAs of NMF in group RJ1 were higher than in group UV+. Despite the increased levels of PAD3, epidermal levels of hydration, filaggrins, glutamate, and serine in groups RJ2, RJ3, and RJ4 were similar to those in group UV+. Dietary supplementation of RJ1 improves epidermal hydration in parallel with enhanced expression and degradation of filaggrin, but not by increased protein expression of PAD3, along with increased generation of glutamate and serine.

Royal jelly enhances migration of human dermal fibroblasts and alters the levels of cholesterol and sphinganine in an in vitro wound healing model

Oral administration of royal jelly (RJ) promotes wound healing in diabetic mice. Concerns have arisen regarding the efficacy of RJ on the wound healing process of normal skin cells. In this study, a wound was created by scratching normal human dermal fibroblasts, one of the major cells involved in the wound healing process. The area was promptly treated with RJ at varying concentrations of 0.1, 1.0, or 5 mg/ml for up to 48 hrs and migration was analyzed by evaluating closure of the wound margins. Furthermore, altered levels of lipids, which were recently reported to participate in the wound healing process, were analyzed by HPTLC and HPLC. Migration of fibroblasts peaked at 24 hrs after wounding. RJ treatment significantly accelerated the migration of fibroblasts in a dose-dependent manner at 8 hrs. Although RJ also accelerated the migration of fibroblasts at both 20 hrs and 24 hrs after wounding, the efficacy was less potent than at 8 hrs. Among various lipid classes within fibroblasts, the level of cholesterol was significantly decreased at 8 hrs following administration of both 0.1 ug/ml and 5 mg/ml RJ. Despite a dose-dependent increase in sphinganines, the levels of sphingosines, ceramides, and glucosylceramides were not altered with any concentration of RJ. We demonstrated that RJ enhances the migration of fibroblasts and alters the levels of various lipids involved in the wound healing process.

Vitamins B1, B2, B6 and PP contents in royal jelly

Hydrosoluble vitamins B1, B2, B6 and PP are essential organic substances for human organism, functioning as coenzymes on several metabolic cycles. In the present investigation four vitamins of B complex and its vitamers contents were determined in royal jelly samples marketed in São Paulo, Brazil. A single extraction process was employed, and each vitamin was determined by HPLC using C18 column and detected by fluorescence. Four samples from different suppliers were analyzed, and the results varied from 0.08 to 0.41 mg/100g (vitamin B1 orthiamine); from 0.01 to 0.05 mg/100g (vitamin B2 or ribofl avin); from 0.13 to 0.38 mg/100g (piridoxal - vitamin B6); from 0.26 to 1.38 mg/100g (piridoxamine - vitamin B6); from 0.21 to 0.57 mg/100g (niacin - vitamin PP);and from 1.56 to 2.00 mg/100g (niacinamide - vitamin PP). These data show that royal jelly is not an important source of the analyzed vitamins, though the results indicate that the employed technique is suitable for determining these four vitamins and its vitamers.

Parâmetros morfofisiológicos testiculares de camundongos (Mus musculus) suplementados com geleia real / Morphophysiological parameters of mice (Mus musculus) testicles supplemented with royal jelly

Avaliaram-se os efeitos da geleia real sobre os parâmetros morfofisiológicos testiculares de camundongos (Mus musculus). Utilizaram-se 57 machos Swiss, com quatro meses de idade, distribuídos aleatoriamente em seis tratamentos: T1: solução fisiológica, via intraperitoneal; T2: 0,1mg de geleia real, via intraperitoneal; T3: 0,2mg de geleia real, via intraperitoneal; T4: água destilada, via oral; T5: 0,1mg de geleia real, via oral; e T6: 0,2mg de geleia real, via oral. Após 45 dias de suplementação com geleia real, os animais sacrificados e pesados tiveram seus testículos coletados, incluídos em parafina e corados com hematoxilina/eosina. Não houve diferença entre os tratamentos quanto aos: pesos corporal e testicular, índice gonadossomático, diâmetro tubular, altura do epitélio, comprimento total dos túbulos seminíferos, comprimento tubular por grama de testículo, índices tubulossomático e leydigossomático e valores de proporção volumétrica referentes à túnica própria, epitélio seminífero, vaso sanguíneo e vaso linfático. Foi encontrada diferença entre T1 e T3 em relação aos túbulos seminíferos e ao espaço intertubular.

The effects of royal jelly on the morphophysiological parameters of mice (Mus musculus) testicles were studied. Fifty-eight male Swiss mice were evaluated. They were four-month old and were randomly distributed in six treatments: T1: physiological solution, intraperitonial route; T2: 0.1mg of royal jelly, intraperitonial route; T3: 0.2mg of royal jelly, intraperitonial route; T4: distilled water, orally; T5: 0.1mg of royal jelly, orally; and T6: 0.2mg of royal jelly, orally. After 45 days of supplementation with royal jelly, the animals were weighted, slaughtered, and the testicles collected, included in paraffin, and stained with haematoxylin-eosin. No differences among treatments were observed for: body and testicular weights, gonadossomatic index, tubular diameter, epithelial height, total length of seminiferous tubules, tubular length per testicle gram, tubulossomatic and leydigossomatic indexes and the value of volumetric proportion related to tunic, seminiferous epithelium, blood vessel, and lymphatic vessel. Differences between T1 and T3 about the seminiferous tubules and intertubular space were found.

Preparation and stability test of Lyophiled Royal Jelly Soft Capsule / 中成药

AIM:To prepare the Lyophiled Royal Jelly Soft Capsule and study its stability and Influential factors.METHODS:The suspending agent and processing method were optimized using sedimentation volume rate as the index.Soft capsules were prepared and product stability under high temperature and high humidity environment was studied according to the determination of the content of 10-HAD by HPLC.RESULTS:The finished product yield in pilot test was more than 90%,the soft capsule products stored in cold were stable,while those stored under room temperature or high temperature and high humidity were unstable with a noticeable decrease in quality.Water content in capsule shell affects the 10-HDA content of the finished product.CONCLUSION:The preparative process is feasible and the products should be storaged in cold enviroment.

Determination of the Levels of 10-HDA in Royal Jelly Preparations Using TLC-UV Spectrophotometry / 中国药房

This paper reports the determination of 10-hydroxy-trans-△~2-decenoic acid(10-HDA) levels in royal jelly preparations using TLC-UV spectrophotometry. This method was accurate, fast and simple with a CV of 12. 7%, offering a tool for drug control. It can be used in qualitative of quantitative analysis of 10-HDA in any royal jelly preparation. Thirty-nine royal jelly preparatons were determined and the results showed that 10-HDA was found only in 51. 3% of them. It seems to be necessary to work out a standard for qualitative identification and quantitative determination of 10-HDA in royal jelly prepartions.

POTENTIATING EFFECTS OF ROYAL JELLY ON IMMUNO LOGICAL FUNCTION IN MICE / 中国药理学通报

Royal jelly (2.5g/kg/d igX8d) increased the clearance rate of iv charcoal particles in normal mice, and completely antagonized the decrease of clearance rate caused by cortisone acetate. It also potentiated the delayed type hypersensitivity ( DTH ) reaction of footpad induced by SRBC in normal mice, and entirely antagonized the inhibition of DTH reaction caused by eye lophosphamide. Royal jelly (2.5 g/kg/d ig X 7 d ) had no marked influence on hemolysin formation in normal mice and immunosuppressed mice caused by eye lophosphamide.

Experimental study of the inhibitory effects of biotic royal jelly on the growth of ascitic hepatoma cell H_( 22) in mice / 第二军医大学学报

Objective: To observe the inhibitory effects of biotic royal jelly on the ascitic hepatoma cell H 22 . Methods: Mice bearing H 22 tumor were fed on different types of royal jelly: No.1, 2 and 3. Their anti tumor effects were observed in vivo . The general royal jelly and normal saline were observed as control. Results: Among these biotic royal jelly, the biotic royal jelly No.1 showed obvious tumor inhibiting and survival prolonging effects. In addition, it increased the number of WBC and augmented the amount of IL 2 and IFN ?; the pathological study also indicated the denaturing and necrosis of most tumor cells with nuclei constraining and cell membrane rupturing, and large amount of lymphocytes and plasmacytes infiltrating around the mass. Conclusion: The No.1 biotic royal jelly has obvious anti tumor effect, and it may take effects by inhibiting or killing tumor cells and improving the immunity of the host. [