Enhancing sperm cryo-preservability, ultrastructure, and testicular histology in rams through oral royal jelly administration

The aim of this study was to investigate the effects of royal jelly against gamma-radiation induced oxidative damage in liver and lung tissue after total body irradiation.The adult male Sprague Dawley rats were randomly divided into six groups of sixteen animals each; group 1: control group (C), group 2: only irradiated rats (IR), group 3: irradiated rats with royal jelly administration at 25 mg/kg/day (IR+RJ25), group 4: irradiated rats with royal jelly administration at 50 mg/kg/day (IR+RJ50), group 5: only royal jelly administration at 25 mg/kg/day (RJ25), group 6: only royal jelly administration at 50 mg/kg/day (RJ50).Royal jelly (RJ) was administered at a dose of 25 and 50-mg/kg body weight, by gavage for 10 days prior to irradiation and 10 days after irradiation.On the tenth day of study, radiotherapy was applied to the whole-body by single fraction at a dose of 6 Gy.Half of rats were sacrificed at 24 hours and 10 days after irradiation under ether anesthesia.Blood samples were collected and analysed for alanine aminotransferase, aspartate aminotransferase, triygliceride, total cholesterol and gamma glutamyl transpeptidase levels.The lung and liver samples were stored for the measurement of malondialdehyde, glutathione peroxidase, superoxide dismutase and catalase activities.Rats exposed to wholebody irradiation induced a marked liver failure, characterized with a significant increase in serum AST, ALT, cholesterol and triglyceride concentrations, and also they had higher lung and liver MDA and lower GSH-Px, CAT and SOD (p<0.001).Administration of royal jelly resulted in a significant decreased in oxidative stress parameters and biochemical parameters, and certainly increased antioxidant activities.Furthermore, pre-and post-treatment with RJ was more effective than pre-treatment with RJ.These results suggest that royal jelly may probably protect the animals from radiation-induced liver and lung damage.The effects of royal jelly can result from its antioxidant properties.

Royal jelly supplementation in semen extender enhances post-thaw quality and fertility of Nili-Ravi buffalo bull sperm

Royal Jelly improves the sperm parameters of ram semen during liquid storage and serves as an antioxidant source

Background and objectiveTo our knowledge, no previous studies have focused on the immunomodulatory effects of fresh royal jelly (RJ) administration on systemic lupus erythematosus (SLE) in humans. Our aim was to study the effect of fresh RJ administration on the disease course in children with SLE with some immunological markers (CD4+ and CD8+ regulatory T cells and T lymphocytes apoptosis).MethodsThis was an open-label study in which 20 SLE children received 2 g of freshly prepared RJ daily, for 12 weeks.ResultsThe percentages of CD4+ CD25+high FOXP3+cells (CD4+ regulatory T cells) and CD8+CD25+high FOXP3+cells (CD8+ regulatory T cells) were significantly increased after RJ treatment when compared with baseline values. Apoptotic CD4 T lymphocytes were significantly decreased after RJ therapy when compared with baseline values and the control group.ConclusionThis is the first human study on the effect of RJ supplementation in children with SLE. Our results showed improvements with 3-month RJ treatment with regard to the clinical severity score and laboratory markers for the disease. At this stage, it is a single study with a small number of patients, and a great deal of additional wide-scale randomized controlled studies are needed to critically validate the efficacy of RJ in SLE.

The current study was carried out to investigate the protective effects of royal jelly supplementation on sperm motility, viability and pH value during the liquid storage of boar semen at 16 °C and 4 °C, at various periods of time (0, 24, 48, 72 and 96 h). Semen samples were collected from 11 boars, diluted with a long-term extender and supplemented with different concentration of royal jelly (0%, 0.5%, 1% and 2%) at a final concentration of 50 × 106sperm/ml. In the laboratory, the semen was assessed for sperm morphology, viability (eosin-nigrosin staining), subjective motility and objective sperm motility by sperm class analyzer. In total, 396 tests for sperm viability and motility were performed. The longer storage time and the lower incubation temperature showed lower sperm motility and viability results. The results showed that royal jelly supplementation at 1% concentrations protected the functionality of the sperm plasma membrane during the liquid storage time of 96 h at 16 °C. Sperm subjective and objective motility results in samples stored at 4 °C decreased with higher royal jelly concentrations and longer storage time, and differ significantly from the results in samples stored at 16 °C (P&lt; 0.05). Our data showed that royal jelly supplementation at lower concentrations can improve boar semen motility and viability parameters during liquid storage at 16 °C for 96 h.

Objective: Royal jelly (RJ) from honeybees (Apis mellifera) has estrogenic activity. Estrogen deficiency after menopause leads to a high risk of memory impairment and depression as well as metabolic syndrome and osteoporosis. We here investigated the effect of RJ on memory impairment and depression-like behaviors in ovariectomized (OVX) rats.Methods: OVX rats were administered with RJ for 82 days. Hippocampus-dependent spatial memory and depression-like behaviors were assessed by the Morris water maze test and the forced swimming test, respectively. The weights of body, brain and uterus and the contents of protein and myelin galactolipids including galactosylceramide and sulfatide were measured.Results: Memory impairment and depression-like behaviors in OVX rats were recovered to the levels of sham-operated rats by RJ administration. Increased body weight and decreased uterine weight in OVX rats were recovered to the levels of sham-operated rats by 17β-estradiol (E2) administration but not by RJ administration. In contrast, brain weight was slightly increased by RJ administration but not by E2 administration. The contents of protein and myelin galactolipids were higher in the brains of RJ-administered OVX rats than in the brains of E2-administered OVX rats.Conclusion: The results suggest that RJ has a beneficial effect on neurological symptoms of a menopausal disorder.

Royal jelly (RJ) is one of the furthermost valuable curative products mentioned by natural medicine scientists due to its promising medical and nutritional purposes. It possesses many impacts, including antioxidants, antimicrobial, antitumor, anti-inflammatory and immunomodulatory functions in human and animal that benefit their health and welfare, resulting in its widespread use in medical and commercial products and healthy food. Recently, favourable functions of RJ on male fertility have been reported in different animals. According to earlier literatures, the level of RJ supplementation in animal diet ranged from 100 to 200mg/kg. Oral exposure to RJ has been reported to have oestrogenic influences in the adult female rats. Also, RJ may be influential in improvement of pregnancy and lambing rates of ewes. Oral administration of RJ at 100mg/kg diet before sexual maturity enhanced sexual behaviour and semen quality of male rabbits. Moreover, RJ administration (up to 400mg/kg diet once weekly) for male rabbits exposed to heat stress can counteract "summer infertility" and improve physiological responses. Furthermore, supplementation of freezing extender media with 0.1 or 3% RJ had a protective influence on cryopreserved and chilled spermatozoa of buffalo and ram respectively. However, the high dose of RJ oral administration (800mg/kg) by pubescent male rats for 1month had an undesirable effect on the reproductive system; however, the somewhat unfavourable influences were mitigated by the discontinuation of the administration. This review shows the chemical composition, favourable applications and health benefits of RJ and its effects on reproductive aspects, semen quality and in vitro fertilization outcomes which are advantageous for scientists, researchers, nutritionists, physiologists, embryologists, pharmacists, veterinarians, pharmaceutical industries and animal's breeders.

Royal jelly (RJ) is a natural bee product that has been used for therapeutic purposes since ancient times. The therapeutic properties of this product, which has rich biological content, are still being investigated with new approaches. In this study, the effect of RJ on telomere length, some antioxidant parameters, and lipid profile was examined. This study will contribute to the literature as it is the first to evaluate the effect of RJ on the length of telomeres in damaged liver tissues. In the study, the levels of serum triglyceride, total cholesterol (TC), high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDL-C), aspartate transaminase (AST), alanine transaminase (ALT), telomerase, 8'-hydroxy-2'-deoxyguanosine (8-OHdG), and paraoxonase-1 (PON1) were investigated with enzyme-linked immunosorbent assay method and telomere lengths were investigated by real-time quantitative polymerase chain reaction. The increased TC, LDL-C levels, and AST and ALT activities in the serum after carbon tetrachloride (CCl4) administration approached the control level after RJ administration. PON1 activity decreased in groups with CCl4. PON1 activity increased after RJ administration. The level of 8-OHdG, which increased groups with CCl4, decreased after RJ administration. According to the results of telomere length analysis in liver tissues, telomere lengths in damaged tissues were significantly shortened with CCl4 application and increased with RJ application. Based on the findings of the study, it was concluded that RJ may have therapeutic effects on telomere lengths and some biochemistry parameters.

Objective(s):Paclitaxel is a potent chemotherapy agent with severe side effects, including allergic reactions, cardiovascular problems, complete hair loss, joint and muscle pain, which may limit its use and lower its efficiency. The cardioprotective effect of royal jelly was investigated on paclitaxel-induced damages.Materials and Methods:Adult male Wistar rats were divided into control and test groups (n=8). The test group was assigned into five subgroups; 4 groups, along with paclitaxel administration (7.5 mg/kg BW, weekly), received various doses of royal jelly (50, 100, and 150 mg/kg BW) for 28 consecutive days. The last group received only royal jelly at 100 mg/kg. In addition to oxidative and nitrosative stress biomarkers, the creatine kinase (CK-BM) level was also determined. To show the cardioprotective effect of royal jelly on paclitaxel-induced damages, histopathological examinations were conducted.Results:Royal jelly lowered the paclitaxel-elevated malondialdehyde and nitric oxide levels in the heart. Royal jelly could also remarkably reduce the paclitaxel-induced cardiac biomarker of creatine kinase (CK-BM) level and pathological injuries such as diffused edema, hemorrhage, congestion, hyaline exudates, and necrosis. Moreover, royal jelly administration in a dose-dependent manner resulted in a significant (P<0.05) increase in the paclitaxel-reduced total antioxidant capacity.Conclusion:Our data suggest that the paclitaxel-induced histopathological and biochemical alterations could be protected by the royal jelly administration. The cardioprotective effect of royal jelly may be related to the suppression of oxidative and nitrosative stress.

Vincristine (VCR) is a chemotherapeutic agent classified as a vinca alkaloid. Royal jelly (RJ) is a significant bee product produced by worker bees, characterized by its high protein content. This study aims to investigate the protective effects of RJ against VCR-induced liver damage. VCR was intraperitoneally administered at a dose of 0.1 mg/kg body weight (b.w.) and RJ was orally administered at doses of 150 and 300 mg/kg b.w. Both treatments were applied to the rats on days 1-6 and 9-14. The composition of RJ was analyzed using LC-MS/MS, revealing the presence of 15 different phytochemical compounds with strong antioxidant properties. Serum samples obtained from the rats were analyzed for ALT, ALP, and AST levels. While these enzyme levels were significantly elevated in the VCR group, a notable reduction was observed following RJ administration. Additionally, SOD, CAT, GPx, and GSH antioxidant parameters, along with MDA levels, were evaluated in liver tissue samples. The results indicated a decrease in SOD, CAT, GPx, and GSH activities/levels and an increase in MDA levels in the VCR group. Furthermore, ELISA was used to assess JAK2, STAT3, and mTOR/PI3K/AKT signaling pathways. VCR administration led to a decrease in mTOR/PI3K/AKT levels and an increase in JAK2 and STAT3 levels. In addition, the mRNA transcription levels of inflammation (NF-κB, TNF-α, and IL-1β), endoplasmic reticulum (ER) stress (IRE-1, GRP78, PERK, and ATF-6), and autophagy markers (LC3A and LC3B) were examined. A significant increase in inflammation, ER stress, and autophagy-related markers was observed in the VCR-treated group. Lastly, the protein expression levels of Bax, Bcl-2, Caspase-3, and NF-κB were evaluated. VCR treatment increased Bax, Caspase 3, and NF-κB levels, whereas Bcl-2 levels were decreased. However, following RJ administration, all these parameters were reversed, demonstrating significant improvements. In conclusion, these findings suggest that RJ may exert a protective effect against VCR-induced liver damage.

ABSTRACTWe investigated microtubule-associated protein 2 (MAP-2) immunoreactivity of hippocampal neurons and the potential role of royal jelly (RJ) in regulating MAP-2 during experimental hypothyroidism (HT). Thirty adult female Wistar albino rats were randomized into five groups: the control group was untreated, the sham control group was treated with 10 mg/kg 0.9% sterile saline injected intraperitoneally (i.p.), The RJ group was treated with 100 mg/kg RJ by oral gavage, the HT group was treated with 10 mg/kg propylthiouracil (PTU) i.p. to induce experimental hypothyroidism, and the HT + RJ group was treated with 10 mg/kg PTU i.p. + 100 mg/kg RJ by oral gavage. Oral and i.p. administrations were performed once daily for 20 days. Thyroid stimulating hormone (TSH) and free thyroxine (fT4) levels in the serum were measured biochemically, MAP-2 was measured immunohistochemically in the hippocampus and an immunohistochemical H score was calculated. MAP-2 immunoreactivity appeared in the cytoplasm of neuron cell bodies and dendrites in the hippocampal CA3 and CA1 regions in the control, sham control and RJ groups. MAP-2 immunoreactivity decreased significiantly in the HT group compared to control, sham control and RJ groups. Also, vascular dilation and swollen cells were observed following PTU administration. The degeneration that was observed in the HT group decreased by RJ administration. By contrast, MAP-2 immunoreactivity increased following administration of RJ. Experimental hypothyroidism reduced significiantly MAP-2 immunoreactivity in both the CA3 and CA1 regions and caused degeneration, including edema and vascular dilation, in the hippocampus. RJ increased MAP-2 expression and exhibited a therapeutic effect on the degenerative changes.

The effect of Alpha-lipoic acid (ALA) on lipid peroxidation, catalase activity and post-thaw semen quality of Achai bulls (Bos indicus) was studied. Semen samples (n=28), collected from adult Achai bulls (n=4) at weekly intervals, were diluted with Tris-citric acid egg yolk extender having various concentrations of ALA (0.00, 0.25, 0.50, 1.00 and 2.00mM) and frozen in liquid nitrogen (-196ºC). Post-thaw semen assessment revealed that the lipid peroxidation decreased, while catalase activity increased (P&lt;0.05) in all extenders containing ALA as compared to control, except in extender containing 2.00mM in which catalase activity decreased (P &gt; 0.05) than control. Moreover, post-thaw semen quality parameters i.e., sperm viability and plasma membrane integrity were higher (P&lt;0.05) in extender having 0.25 and 0.50 mM ALA compared to control, whereas mitochondrial membrane potential was significantly improved only with 0.50mM ALA as compared to control and all other ALA concentrations used. However, acrosome and DNA integrity of spermatozoa increased in extender supplemented with 0.25, 0.50 and 1.00mM ALA (P&lt;0.05) compared to control. Computer Assisted Semen Analysis (CASA) of post-thaw semen revealed that total motility (%) and curvilinear velocity (µm/s), progressive motility (%), progressive fast motility (%) and beat cross frequency (Hz), and straight-line velocity (µm/s) and average path velocity (µm/s) were higher (P &lt; 0.05) for 0.50 and 1.00mM, 0.25 and 0.50mM and 0.50 and 2.00mM ALA concentrations, respectively, than control. In conclusion, the addition of 0.50mM ALA in Tris-citric acid, fructose, egg yolk glycerol extender can perform optimum among all ALA concentrations (0.00, 0.25, 1.00 and 2.00mM) used, in terms of minimizing the oxidative stress and improving the post-thaw semen quality of Achai bulls. Keywords: Bos indicus, Achai bulls, Alpha-lipoic acid, Post-thaw semen quality, Antioxidant enzyme, Computer Assisted Semen Analysis.

Restraint and cold stress increase both corticosterone and glycemia, which lead to oxidative damages in hepatic tissue. This study assessed the effect of royal jelly (RJ) supplementation on the corticosterone level, glycemia, plasma enzymes and hepatic antioxidant system in restraint and cold stressed rats. Wistar rats were allocated into no-stress, stress, no-stress supplemented with RJ and stress supplemented with RJ groups. Initially, RJ (200mg/Kg) was administered for fourteen days and stressed groups were submitted to chronic stress from the seventh day. The results showed that RJ supplementation decreases corticosterone levels and improves glycemia control after stress induction. RJ supplementation also decreased the body weight, AST, ALP and GGT. Moreover, RJ improved total antioxidant capacity, SOD activity and reduced GSH, GR and lipoperoxidation in the liver. Thus, RJ supplementation reestablished the corticosterone levels and the hepatic antioxidant system in stressed rats, indicating an adaptogenic and hepatoprotective potential of RJ.

Background: Both coding and long non-coding RNAs (lncRNAs) have emerged as vital regulators in almost every cellular process, and their expression can be modulated by external stimuli, such as physical exercise. Objectives: The current research aimed to investigate the effects of different volumes of TABATA-high-intensity interval training (HIIT) exercises combined with royal jelly (RJ) supplementation on the NLRP3 inflammasome and lncRNA-H19 expression in obese males. Methods: Forty-two healthy men [Body Mass Index (BMI) = 30 kg/m², waist-to-hip ratio = 0.95, age range: 40 - 60 years] volunteered to participate in the study. The individuals were randomly divided into five experimental groups (N = 35) and one control + placebo group (N = 7). The high-volume (HV) or low-volume (LV) TABATA exercise programs were performed twice a week for 8 weeks. Participants in the RJ supplementation groups received a 1000 mg capsule once a day for 8 weeks. The expression of NLRP3 and lncRNA-H19 genes was evaluated using the real-time PCR method. Results: The NLRP3 gene expression in the Bruce test, measured before and after the 8-week exercise interventions and RJ supplementation, showed insignificant changes across the different groups. However, the H19 gene expression in the Bruce test showed a significant reduction in the HV-TABATA HIIT intervention groups, which was more pronounced than in the LV groups after 8 weeks: HV group (P = 0.004), RJ group (P = 0.001), HV + RJ group (P = 0.007), and LV + RJ group (P = 0.002). After 8 weeks of non-pharmacological interventions involving exercise training and supplementation, a significant decrease in NLRP3 and a significant increase in H19 gene expression were detected in the HV group compared to the LV group (P = 0.05 and P = 0.010). Significant improvement was also found in the resting H19 levels between the RJ and LV groups (P = 0.011) and the LV + RJ group (P = 0.44). Moreover, a significant reduction in resting NLRP3 gene expression was observed between the RJ + LV and LV groups (P = 0.038). Conclusions: Chronic HV TABATA HIIT exercise, when combined with RJ supplementation, is effective in attenuating inflammatory responses to acute stress.

Objective: Royal jelly (RJ) has been used for medical and nutritional purposes, and previous studies have indicated that it may have estrogenic activity. The present study investigated the effects of RJ on bone metabolism in ovariectomized (OVX) rats.Methods: Rats (12 weeks old) were randomly divided into four groups, namely Baseline, Sham, OVX, and OVX + RJ groups. Rats in the Baseline group were killed immediately, whereas rats in the OVX and OVX + RJ groups underwent bilateral ovariectomy and those in the Sham group underwent sham operation. RJ was administered to rats in the OVX + RJ group daily for 12 weeks. At the end of the 12-week period, bone mass, bone histomorphometry, and bone mechanics were analyzed.Results: Femur bone mineral density (BMD) was significantly lower in the OVX group than in the Sham group, and this decrease in BMD was not ameliorated by RJ administration. However, femur stiffness, as evaluated by a three-point bending test, was significantly higher in the OVX + RJ group than in the OVX group.Conclusion: The findings of the present study suggest that RJ does not prevent bone loss, but does improve bone strength in OVX rats.