Apimondia 2015 Abstract book (South Korea)

screened as arginine-induced insulin secretagogues. Brazilian propolis induced insulin secretion higher than arginine did. Surprisingly Chinese propolis had no activity for insulin production.

Propolis is a resinous mixture that bees collect from tree buds, sap flows, or other botanical sources, suggested that the contents of propolis are different from the places. We focused Drupanin and Artepillin C, which are small chemical compounds and found in Brazilian propolis specifically. They had some insulin secretion activity, similar to arginine, indicated that there were some other factors, which were the arginine mimetics for insulin secretion.

To identify target factor(s) of arginine/Brazilian-propolis induced insulin secretion, our previous study was that nanobeads technology facilitated us to purify chemical-target factors. This time we chose the other way, insulinassociated factor purification. Two insulin-associated factors were purified and identified.

Latest results will be present and discussed.

ATO-026

Anti-parasitic activity of propolis

Eman AbdelRahman, Ahmed Hegazi

National Research Centre, Egypt

Propolis (bee glue) is a resinous hive product, collected from various plant sources. It has been long used in folk medicine of different nations as early in Egypt as 3000 BC. It has attracted much attention as a useful material applied in medicine due to its pharmacological and biological activities. Researchers have been interested in the investigation of isolated compounds responsible for propolis action; since propolis-containing products have been marketed and humans have used propolis for different purposes. The efficacy of propolis in different protocols in vitro and in vivo suggests its therapeutic properties. Recently, attention is being focused on the antiparasitic activity of propolis, the goal of this review is to discuss the potential of propolis for the development of new drugs, by comparing data from the literature that suggest candidate areas for the establishment of drugs against parasites.

ATO-045

Biological activity of Artepillin C obtained from Brazilian green propolis

Byungyoon Cha

CHUBU University, Japan

Brazilian green propolis, which is produced by bees that collect exudates from Baccharis dracunculifolia DC in southern and southeastern regions of Brazil, contains high levels of Artepillin C (ARC) and other flavonoids. Studies have demonstrated that ARC possesses antimicrobial, antioxidant and antitumor activities. ARC also exhibits anti-inflammatory effects in vitro and in vivo. In our previous study, we demonstrated that ARC induces adipocyte differentiation and stimulates glucose uptake in 3T3-L1 adipocytes by activating peroxisome proliferator-activated receptor (PPAR) as a ligand (Choi SS et al. 2011. Biochem Pharmacol.81:925-33). In the present study, we investigated the comprehensive effects of ARC on metabolic functions in the diabetic obese ob/ob mouse model, and in a co-culture system of adipocytes and macrophages. We demonstrated that ARC suppressed the production of proinflammatory mediators such as monocyte chemotactic protein 1 (MCP -1) and

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interleukin 6 (IL-6), and decreased the development of insulin resistance by regulating inflammatory interactions between adipocytes and macrophages in adipose tissue. ARC also blocked the expression of these proinflammatory mediators in TNF-stimulated adipocytes and fatty acid-stimulated macrophages. In a rodent model of diabetes and insulin resistance (ob/ob mice), ARC ameliorated abnormalities in plasma glucose and lipid levels. In vivo, ARC reduced the level of inflammatory cytokines. These results indicate that ARC may have beneficial effects for the prevention and/or treatment of insulin resistance.

Symposium: Clinical Studies of Bee Venom

ATO-006

Treatment of neuralgia with microdoses of bee venom

Cristina Pavel

Romanian Apitherapy Society, Romania

Bee venom is powerful analgesic and can replace successfully classic medication for pain management. The paper presents 6 cases of neuralgia (trigeminal neuralgia, lombosciatica, acute lumbago, pudendal neuralgia and lumbar hernia 2 cases) which were helped by only few sessions of bee venom in microdoses. In all cases, patients tried before several other treatments, even surgery (for lumbar hernia), but pain persisted. Because of acute pain, the doses of bee venom were very small, otherwise pain would have been aggravated. The advantage of bee venom treatment is very fast but also long lasting therapeutic response.

ATO-029

Effect of Bee venom acupuncture as a complementary modality for treatment of chronic low back pain

Maha Saber, Eitedal Daoud, Aliaa al Gendyl, Khaled Abdel-Wahhab, Ahmed Hegazi

National Research Centre, Egypt

Bee venom acupuncture (BVA) from honeybee (Apis mellifera) was assessed for their potential use as a complementary modality for treatment of chronic low back pain (CLBP). The purpose of this study is to determine whether BVA is effective at improving pain intensity, functional status and quality of life in patients with CLBP. This study is a randomized, controlled clinical trial with two parallel arms. We intend to compare the effects of BVA by bee sting or bee venom collected by the electric shock device and pharmacotherapy with pharmacotherapy only in patients with CLBP Patients with chronic LBP will be recruited with a target sample size of 40 participants (18 and 65 years of age, experiencing LBP lasting for at least the previous three months, scoring 4 points on a 10-cm visual analog scale (VAS) of LBP at the time of screening). The results revealed that the bee venom apiacupuncture showed significant improvement in patients received bee venom group compare to patients received pharmacotherapy only. It is concluded that both modes of treatment for CLBP gave improvement regarding pain intensity, disability and quality of life being more evident in bee venom group supported with improved serum (IL1 and NF-KB).

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ATO-034

Apitherapy in diabetes

Vasily Krylov, Anna Deriugina, Olga Barinova

Lobachevsky State University of Nizhny Novgorod, Russia

The paper investigated the effects of bee products for the maintenance of blood glucose levels when modeling rats diabetes 1 and 2 types. Experimental modeling of diabetes was conducted on 70 rats. Type 2 diabetes in animals caused per os administration of 40% glucose for 14 days. Type 1 diabetes cause ip administration of alloxan (130 mg / kg). Therapy bee products (propolis (per os 100 mg / kg), royal jelly (per os 100 mg / kg), bee venom, (i p 0,1 mg / kg) was performed 10 days. Control animals were simulated in which the diabetes without further apitherapy and rats not exposed. The results showed that in type 1 diabetes glucose in the blood of rats was 32,47 ± 1,08 mmol / l, with type 2 diabetes - 25,15 ±1,92 mmol / l, in the normal - 5,22 ± 0.21 mmol / l. Apitherapy defined decrease in blood glucose in rats. Propolis reduced concentration of glucose in the blood in type 2 diabetes by 50% in a week, royal jelly by 58% relative to the control group. With type 1 diabetes a decrease in concentration of glucose in the blood for a group of "propolis" - 38% "royal jelly" - 41%. The maximum decrease of glucose recorded in animals injected ip bee venom. A week before the rate was restored to normal in type 2 diabetes, and approaches to normal in the group with type 1 diabetes. In control rats, there was no spontaneous lowering blood glucose.

ATO-044

Anti-wrinkle effects of honeybee venom serum on facial wrinkles

Sangmi Han1, Sungnam Chun2, Kwankyu Park3, Young mee Nichollos4, Sokcheon Pak5

1 National Academy of Agricultural Science, Republic of Korea

2 DongSung Pharm. Co. Ltd

3 School of Medicine, Catholic University of Daegu

4 Manuk Doctor Ltd

5 Charles Sturt University

Acial wrinkles are an undesirable outcome caused by extrinsic photodamage and intrinsic aging process. Currently, no effective strategies are known to prevent facial wrinkles. The main aim of the present study was to evaluate the clinical effects of bee venom serum by measuring the clinical wrinkles objectively with a device that converts the wrinkle roughness to numerical values. This is the first clinical study to determine the effects of bee venom containing cosmetics on facial wrinkles in human skin. The main aim of this study was to evaluate the clinical effects of bee venom serum. We analyzed images using replica and a device of Visioline which is an objective technique to reproduce changes in photodamaged skin. The image analysis using replicas was performed at week 12 when significant differences in total wrinkle area, total wrinkle count and average wrinkle depth were observed. This analysis correlated well with the clinical findings. Moreover, the significant difference in visual grade was observed after 8 weeks with bee venom serum indicating the faster effect of bee venom serum on wrinkle improvement. Our findings suggest that bee venom plays an important role in the improvement of skin wrinkles which is used in the cosmetics industry as an anti-wrinkle agent. Bee venom serum provided a greater efficacy in terms of total wrinkle area, total wrinkle count and total wrinkle area in subjects with photodamaged skin. Longterm treatment with bee venom containing cosmetics could be safe because the irritation potential of bee venom is negligible.

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ATO-071

Comprehensive and functional venomics of social wasps

Vespa crabro flavofasciate Cameron and Vespa analis parallela Andre

Kyungjae Yoon1, Kyungmun Kim1, Phuong Nguyen2, Jong Bok Seo3, Young Han Park4, Ki-Gyoung Kim5, Hong-Yul Seo5, Young Ho Koh2, Si Hyeock Lee1

Vespa crabro is a cosmopolitan social wasp species whereas Vespa analis is commonly found in Asia. Both species are widely distributed in Korea and known to be aggressive when disturbed, resulting in frequent sting accidents. Although major venom components of well-known Vespa wasps have been reported, no comparative transcriptomic analysis of venom gland between V. crabro and V. analis has been conducted to date. To investigate the differences in venom properties between these two wasps, total RNA was extracted from each venom gland and used for RNA-sequencing. A total of 36 and 37 venom-specific genes were identified in venom gland transcriptomes from V. crabro and V. analis and their expression profiles were different.Prepromastoparan A, Vespid chemotactic precursor and vespakinin-T were the top three genes that were most prevalently transcribed in the venom gland of V. crabro, and their transcription rates were 112-, 16and 161-fold higher compared with V. analis, respectively, as judged by FPKM values. Their differential transcription profiles were confirmed by quantitative real-time PCR. Our findings suggest that V. crabro produce venom with much more enriched venom components, thereby resulting in higher toxicity compared with V. analis. In an attempt for the functional analysis of venom components, mastoparans of both V. crabro and V. analis were synthesized and their bio-activities were compared. The V. analis mastoparan exhibited similar levels of antimicrobial activities against model bacteria and fungi but significantly higher antitumor activities against human ovarian tumor cell lines compared with the V. crabro mastoparan.

ATO-062

Bee bread–Various usage of bee bread and clinical experiences

Pushpendra Singh Bhandari1, Mirela Strant2

1 Member National Bee Board Of India, Member of German and Romanian Apitherapy Societies,

Jaipur, India

2 FM licenced, Trainer in Api-Phyto-therapy, Cluj-Napoca, Romania

Bee bread is the pollen which the bees process in their hives.There it suffers a process of fermentation in a stable condition of temperature and humidity, wich gives it some different characteristic. For example, bee bread contains more reducing sugars than pollen. It has higher acidity due to the presence of lactic acid and larger amount of vitamin K. The quality of lactic acid is by sixfold higher as compared with pollen. Also, bee bread contains a milk-digesting enzyme which pollen does not. Another important thing to be mentioned is that the outer

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shell of pollen (exine) is broken during this process, releasing nutrients. All these factors makes from bee bread a precious and valuable product which can be used in medical practice and also in our diet. Bee bread has high antioxidant and scavenging abilities against free radicals. At present, consumer’s demand for natural foods with the effect of a medicine is increasing. Bee bread is a food containing naturally occurring compounds. In the future, bee bread will apply more and more as health food and medicine due to its functional properties. In the final of our presentation we present some clinical cases and experiences using bee bread as remedy and superfood as well.

Symposium: Pharmacological Effects of Bee Venom

ATO-032

Integrated apitherapy treatment of chronic osteomyelitis

Siu Wan Ip, Yui-Chung Chang

Asia Pacific Apitherapy Association, Taiwan

PURPOSE: To investigate the effects of integrated medical treatment of Apitherapy (Bee venom Treatment),bee propolis, royal jelly and Medical Nutrition Therapy(MNT) on chronic Osteomyelitis patient. METHOD: A 22- year-old female motorcycle rider had a severe traffic accident in Chang-Hua City, TAIWAN. Her left leg was hurt by a vehicle and broken into three parts with open wounds fractures. Patient was admitted to Taiwan Chang-hua Christian Hospital. Amputation of left leg was advised because of severe soft tissue infections spreading to bone. She refused for amputation and she was transferred to National Tri-Service Military Hospital (NTMH). After debridement of necrotic bone and prolonged nine months administration of antibiotics, she was discharged from NTMH. However, she found that she had chronic Osteomyelitis. Purulent fluids was found over left leg in every summer. She visited our Apitherapy clinic and received three times of BVT weekly combined with high protein diet Medical Nutrition Therapy (MNT), bee propolis and royal jelly. RESULT: After received six months of integrated Apitheray treatment, she found that she do not had purulent fluids in her leg. CONCLUSION: Integrated Apitherapy with BVT, bee propolis and royal jelly combined with MNT with high protein diets during the course of Apitherapy can increase the effect of Bee venom Treatment in chronic Osteomyelitis.

ATO-031

Hypolipidemic and blood glucose lowering activity of honey bee venom (Apis mellifera) in alloxan induced diabetic rabbits

Solongo Ganbold3, Khulan Tserennadmid1, Uuganbayar Baatartsogt2

1 Mongolian National University of Medical Science, Mongolia

2 Institute of Traditional Medicine and Technology

3 Shidet Zugii, Mongolia

Introduction: In diabetes therapy, a great attention is paid on lowering blood glucose levels and lipid regulating mechanisms of various medical agents including animal toxins. Honey bee venom (BV) reduces blood glucose level through increased insulin secretion and glucose take-up.BV also has lipid regulating activity verified in

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several other studies. Thus bee venom could be considered as a potential remedy for diabetes. This study aims to investigate the effect of Mongolian honey bee venom on hyperglycemia and hyperlipidemia in alloxan induced diabetic rabbits. Material and Method: Twenty two Chinchilla rabbits were divided into groups: the control (n=6), the diabetic (n=8), and the bee venom treated (n=8) groups. In diabetic group iabets was induced with Alloxan monohydrate. The bee venom treated group received a bee sting (a sting contains 0.2-0.5 ml of bee venom) on their hind paw every other day after the confirmation of diabetes. Result: Bee venom treatment (BVT) led to the following changes: compared to the diabetic group, the bee venom treated group’s blood glucose levels decreased by 14.9% -26.5%; blood cholesterol levels reduced by 12.5%-19.1%; low density lipoproteins (LDL) levels lowered by 11.2%-14.2% ; and high density lipoproteins (HDL) levels increased by 2.5% - 26.25%. Conclusion: Bee venom lowers blood glucose levels and improves lipid profile in alloxan-induced diabetic rabbits and can be considered as a therapeutic agent for diabetes. Further studies should be carried out to determine the most appropriate bee venom dose for the best therapeutic effect.

ATO-030

Effect of Indian honey bee venom (Apis cerana) on wound healing efficacy, anti-bacterial activity and purification of peptide components

Bhargava Hunkunda Radhakrishna

SASTRA University, India

Introduction: The main components of Apis bee venom are melittin, apamin, phospholipase A2, Hyaluronidase and amino acids which have diverse pharmacological properties and biological activities. Melittin has shown significant results against several pathogens. For this reason, bee venom could be considered as a potential antibacterial and anti-inflammatory property. The purification of the peptide compounds can be done by the gel filtration chromatography and SDS PAGE. This study aims to investigate the wound healing, antibacterial and purification of peptide components from Indian honey bee venom. Materials and Methods: Eighteen Wistar rats were divided into three groups (n=6): the control, the standard and the treated groups. The antibacterial activity was determined using disc diffusion. The components were separated from the bee venom by gel filtration chromatography on Sephadex G-50 column and confirmed through SDS PAGE. Results: The wound contraction was superior in animals treated with bee venom with a significant increase in the level of serum albumin than other groups. The histopathological studies revealed confirmatory evidence and enhanced epithelialization in the animals treated with bee venom. The 5mg.mL-1 showed significant results for antibacterial activity. The fractions obtained were melittin, phospholipase A2 and other higher fractions analysed by SDS PAGE. Conclusion: Apitoxin (honey bee venom) can be used as an alternative to treat different kinds of wounds and due to significant anti-bacterial efficacy it can be an alternative to antibiotics and it will have a great impact in the near future.

ATO-008

The regulatory effects of purified bee venom on Propionibacterium acnes-induced inflammatory responses in vitro and in vivo

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Woo Ram Lee, Hyun Jin An, Jung yeon Kim, Hyun Chung, Kwan kyu Park

Catholic University of Daegu, Republic of Korea

Purified bee venom (BV) is a natural toxin produced by honeybees. BV has been widely used as a traditional medicine for vario us diseases. However, the anti-inflammatory mechanisms of BV have not been elucidated in the Propionibactierium acnes (P. acnes) induced keratinocyte or inflammatory skin disease animal models. In this study we examined the effects of BV on the production of inflammatory cytokines in heat-killed P. acnesinduced HaCaT cells. Heat-killed P. acnes treated keratinocytes were increased expression of pro-inflammatory cytokines. However, BV treatment significantly suppressed the expression of these cytokines. Subsequently, we examined the effect of living P. acnes (1×107 CFU) intradermally injected into the ear of mice. Living P. acnes injected ears showed cutaneous erythema, swelling, and granulomatous response at 24 h after injection. However, BV-treated ears showed markedly reduced swelling and granulomatous responses compared with the ears injected with only living P. acnes. These results demonstrate the feasibility of applying BV for the prevention of the progression of inflammatory skin diseases induced by P. acnes in vitro and in vivo inflammatory models.

ATO-039

Bioactive properties of honey bee products and their mixtures

Sibel Silici

Erciyes University, Turkey

Four honey, four bee pollen, propolis, royal jelly and 4 different their mixture (honey, polen, royal jelly and propolis) samples obtained form Turkey were screened for total phenolic content by the modified Folin– Ciocalteu method, for potential antioxidant activity using phosphomolybdenum assay and by the 1,1-diphenyl-2- picryl hydrazyl (DPPH) method for antiradical activity. Total phenolic content of samples ranged from 0.58 to 320.83 mg GAE/100 g honey. Antioxidant activity of samples was between 29.05 and 267.37 mg AAE/g honey. Radical scavenging activities of the samples were varied between 5.72 % and 96.13 %. Propolis, castanea honey, castanea pollen and honey-RJ-bee pollen mixture were the most active among samples.

Symposium: Pharmacological Effects of Royal Jelly, Bee Pollen

ATO-061

The effects of N•Chromosome royal jelly & bee products on diabetes type 2

Hossein Yeganehrad1, sajad Jazani-Dorche1, Maryam Moarefi2

1 Caspian Apiaries, Canada

2 Karaj, Branch, Islamic Azad University, Karaj, Iran

DIABETES TYPE 2 is a chronic condition that effects the body’s natural ability to metabolize glucose (sugar).

This may be the result of insulin resistance, which can prevent the pancreas from making enough insulin or effectively utilizing it. This illness is also known as noninsulin dependent diabetes because insulin is still produced

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within the bodies of individuals with this disease. If a build up of glucose occurs, the cells in the body are unable to function properly. This can lead to protein deficiencies and many other disorders since Diabetes sufferers are unable to effectively utilize protein it leaves them susceptible to many autoimmune disorders. We recently tested a new method of treatment with N-Chromosome Royal Jelly in conjunction with other bee products.On 536 insulin dependent diabetics (70% of which require amputations,) each individual was suffering from at least one other auto immune disorders directly related to the severe symptoms of their condition.After the first 10 weeks of treatment (dependent on severity) amputations were not required in 100% patients and pancreas was able to effectively produce and utilize insulin, therefore insulin injection was no longer required for over 90% of patients after 3 months of treatment. During this presentation we will explain the external & internal use of N-chromosome royal jelly as an anti-inflectional supplementary recovery to treat certain deficiencies, and autoimmune disorders in the body

ATO-048

MRJP-1 multimer activates cell proliferation of fibroblasts originated from gingiva and periodontal ligament

Kikuji Yamaguchi1, Toru Suzuki2, Kiyoshi Murata2, Akira Fujii3, Yoshihisa Yamaguchi4

1 Institute of JRJ Apimedical Science, Japan Royal Jelly Co. Ltd.; Tohoku Fukushi University; Yunnan

Agricultural University, Japan

2 Institute of JRJ Apimedical Science, Japan Royal Jelly Co. Ltd.

3 Institute of JRJ Apimedical Science, Japan Royal Jelly Co. Ltd.; Nihon University

4 Japan Royal Jelly Co. Ltd.

OBJECT: Periodontitis is an inflammatory disease affecting the periodontium, the tissues that surround and support the teeth, such as alveolar bone, periodontal ligament, and gingiva. Significant associations between periodontal disease and cardiovascular disease, diabetes mellitus, preterm low birth weight, and osteoporosis have been discovered. Therefore, the prevention of periodontitis is very important not only oral disease but also severe systemic disease. We have reported that royal jelly inhibits the growth of Porphyromonas gingivalis andcould be applied for periodontitis. The production of connective tissue is one of the important factors to prevent the progression of periodontitis. Therefore, it was very interesting to clarify if MRJP-1 multimer, a major protein component in royal jelly (RJ), affects the cell growth of fibroblasts in periodontium. METHOD: Cells originated from gingiva and periodontal ligament were grown in DMEM-10 supplemented with 10% FCS and antibiotics (streptomycin, penicillin G, amphotericin B) under 5% CO-95% air at 37. Cells were plated in 96-well plate, adjusted growth phase with DMEM-0.5, and then treated in serum-free DMEM supplemented with antibiotics, and RJ or MRJP-1 multimer (0 to 400 /). The cell growth was measured using CCK-8(cell counting kit-8, DOJINDO, Japan) on 2nd, 4th, 6th, and 8th days. RESULTS: Both MRJP-1 multimer and RJ enhanced the cell growth in dose dependent manner in both fibroblasts originated from gingiva and periodontal ligament. This might indicate that RJ and MRJP-1 multimer is useful to strengthen periodontium and prevent the progression of periodontitis.

ATO-004

Royal jelly inhibits the growth of breast cancer in mice

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Zhang Shuang, Su Songkun

Fujian Agriculture and Forestry University, China

Royal Jelly (RJ) is widely consumed in diets in many countries due to its various pharmacological properties: anti-oxidative, antihypertensive, anti-inflammatory, antibiotic-resistant bacteria, etc. We have investigated the role of RJ in the development of breast cancer in mice. Normal BALB/c female mice were injected 4T1 cell at 0.1mL per mouse. The animals orally treated with RJ (0, 0.5, 1.0, or 1.5g/kg Body weight) for 28 days. Internal organizations of the animal were collected for the weight. ELISA assay were used to detect the antioxidant assays of the blood serum, the liver and the kidney; and the immunity of the blood serum. The liver, the kidney and the tumor were taken for histological analyses. Our study showed that the weight of internal organs was different. Treatment with RJ significantly reduced the increase of breast tumor (p<0.05), spleen (p<0.01) and thymus (p<0.01). The antioxidant capacity of the blood serum, the liver and the kidney was increased in mice fed RJ. RJ treatment enhanced the level of immunity of the blood serum. These results corroborated with the RJ supplementation in diets. The antioxidant capacity of RJ may play an important role of anti-tumor growth.

ATO-011

Transylvanian raw pollen – inhibiting effect on murine colon carcinoma culture cells; clinical case

Alina Varadi1, Rodica Margaoan2, Ramona Grosu3, Mirela Strant3, Ioan Cozma4

1 CasaBio with Health, Cluj Napoca, Romania

2 Eng. PhD, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania 3 Dr., CasaBio with Health, Cluj-Napoca, Romania

4 Beekeeper, photographer; Transylvania, Romania

Raw pollen, by its composition and properties is a great food for human being, even a super-food. The pollen is a product made from male reproductive cells of the flowers whose role is to perpetuate the genetic information. Until some years ago, the standard way of preserving the pollen over the years was to dry it off. No one contested the benefits it brought, but dried in heat the lactobacils, bifidobacils, yeast, a large amount of vitamins, enzymes, pro-enzymes and antibiotics properties - most of its active principles are destroyed. To sum up, the experience shows: the raw pollen is highly superior to the dry one. RAW POLLEN is better and more efficient in treating colon and bowel disorders, as much as other illnesses and its attributes are undeniable. Raw pollen is a “living” food, charged with enzymes, friendly bacteria, proteins, minerals, vitamins, antioxidants and a natural delicious taste. Recently, in Veterinary Medicine and Agriculture University from Cluj-Napoca was made an experiment with methanol extract of raw pollen which showed anti tumor effect on murine colon carcinoma by inhibiting cell proliferation and induced apoptosis. Our findings suggest that the effects of polyphenols and flavonoids on colon cancer cells were observed in a dose-dependent manner. On a clinical case, the results suggest that this effect may be eloquent on human being. We tested a recipe for eliminating body toxins based on raw pollen and healthy diet.

ATO-052

Proteomic research on the effect of royal jelly on carbon tetrachloride-induced liver injury in rats

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Miao Jing, Luo Yajuan, Shi Peiying, Wu Zhenhong, Miao Xiaoqing

College of Bee Science, Fujian Agriculture and Forestry University, Fuzhou, China.

Many studies have confirmed that royal jelly has hepatoprotective effect, the purpose of this study is to investigate protective effects of royal jelly against carbon tetrachloride-induced liver injury in rats by proteomics. The hepatic model developed by injection of 15% CCl4 corn oil (2 mL•kg-1) in the first, forth, seventh and tenth day, for 4 times. The level of serum ALT, AST, ALP and MDA in liver homogenates were determined, and the liver histopathology of each group was observed; the different proteins of normal group, model group and treatment group (royal jelly) were analyzed by two-dimensional electrophoresis and MALDI-TOF/TOF-MS. The results of liver histopathology showed that: compared with normal control group, we found edema lesions and severe inflammatory cell infiltration in model group, and these phenomena were improved in each group of royal jelly. In addition, the proteins with significantly different expression were identified by MALDI-TOF/TOF-MS, and 32 proteins were successfully determined. The results showed that most of determined proteins were associated with metabolism (suc h as regucalcin), and some proteins (such as cytokeratin 18, heat shock proteins) that can be used as bio-marker of the protective effect of royal jelly on carbon tetrachloride-induced liver injury in rats. The results revealed the pathways and mechanisms of the protective effect of royal jelly on liver damage in some degree.

ATO-038

Studies on estrogenic activities of royal jelly and its fatty acids in vitro and in vivo

Kenji Ichihara1, Satoshi Uchiyama1, Shou Hotta1, Tsuyoshi Nakanishi2

1 API Co., Ltd., Gifu, Japan

2 Gifu Pharmaceutical University, Gifu, Japan

Royal jelly (RJ) has been used as a nutrition supplement for relief of menopausal symptoms among Japanese women. Previous studies have demonstrated that RJ possesses estrogenic activity in vitro and in vivo. In addition, fatty acids derived from RJ have been reported to modulate estrogen receptor-mediated functions. On the other hand, the stimulation of estrogen receptors might increase the risk of disease exacerbation in patients with breast cancer, uterine cancer (type I), or endometriosis. In this context, we reevaluated the possible estrogenic activity of RJ in vitro and in vivo. The RJ fatty acids such as 10-hydroxy-2-decenoic acid, 10-hydroxydecanoic acid, 2- decene-1,10-dioic acid, and sebacic acid did not show apparent responses associated with the activation of estrogen receptor in a nuclear receptor cofactor assay, in estrogen receptor-mediated gene expression using a luciferase reporter gene assay, and in an estrogen-dependent proliferation assay with MCF-7 human breast cancer cells. The RJ sterol 24-methylenecholesterol enhanced estrogen receptor-mediated gene expression but not the MCF-7 cell proliferation. Then, the in vivo study was conducted using a novel transgenic mouse (E-Rep mouse), which sensitively detects the kinetics of gene activation by estrogenic compounds. The result is that any estrogenic activity could not be found in E-Rep mice fed a diet containing 4% (w/w) freeze-dried RJ for 7 days. Taken together, we concluded that RJ probably does not cause estrogen receptor activation which leads to worsening estrogen-dependent diseases.

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