The protective effects of blueberry honey from Canada against H2O2-induced cytotoxicity in human buccal mucosal cells.

OBJECTIVES: Several clinical trials have been conducted worldwide to evaluate the efficacy of honey against stomatitis. However, it is unclear which types of honey are effective at preventing and/or treating stomatitis. This study aimed to evaluate the potencies of several types of honey in preventing and/or curing aphthous stomatitis in in vitro studies. METHODS: The following experiments were performed: H2O2-induced cytotoxicity and mucosal cell migration in a scratch assay using buccal mucosa squamous carcinoma (HO-1-N-1) cells and the cellular expression of heme oxygenase-1 (HO-1) mRNA encoding an enzyme involved in protection against oxidative stress by real-time RT-PCR analysis, and liquid-liquid extraction and UHPLC analysis in order to examine the active components of honey. RESULTS: Of the 13 types of honey used, Canadian blueberry honey exhibited the protective effect on H2O2-induced cytotoxicity and enhanced cell migration. In addition, blueberry honey increased the expression of HO-1 mRNA with and without cotreatment with H2O2. With regard to active components of blueberry honey, the water-soluble components with a mass of >10 kDa showed a cytoprotective effect, but they have not been identified. CONCLUSION: Canadian blueberry honey, but not the other types of honey, prevents H2O2-induced oxidation of cells, probably through activation of the antioxidant and cytoprotective enzyme HO-1. Blueberry honey also enhanced cell migration, which may be relevant to wound healing. The results of this study suggest the possibility of prophylactic and therapeutic effects of Canadian blueberry honey on human stomatitis that could complement existing treatments.

Royal jelly reduces depression-like behavior through possible effects on adrenal steroidogenesis in a murine model of unpredictable chronic mild stress.

Royal jelly (RJ) is used as a dietary supplement for human health promotion. Recently, a clinical trial has reported that RJ improved mental health. The present study was conducted to experimentally support the clinical effect of RJ on mental health and to further elucidate the mechanisms of action of RJ. RJ and an ethanol extract of RJ, which contains fatty acids but not proteins, inhibited an unpredictable chronic mild stress (UCMS)-induced increase in immobility time, a depression-like behavior, in the tail suspension test. DNA microarray analysis of the adrenal grand revealed that the expression of genes involved in cholesterol metabolism was up-regulated in response to UCMS exposure and that RJ suppressed expression of genes related to cholesterol synthesis and transport. These results suggested that RJ improves stress-induced depression-like behavior by regulating adrenal steroidogenesis and that fatty acids contained in RJ partly contribute to the antidepressant effect of RJ.

Effects of long-term administration of royal jelly on pituitary weight and gene expression in middle-aged female rats.

To determine the effects of ingested royal jelly (RJ) on the pituitary in middle-aged female rats, we performed a long-term RJ administration test. Several animals showed age-related increases in pituitary weight, and RJ administration compensated for the increase. RJ tended to down-regulate prolactin mRNA and up-regulated thyroid-stimulating hormone beta mRNA in the pituitary. This suggests that RJ compensates for age-associated decline in pituitary functions.

Estrogenic activities of Fatty acids and a sterol isolated from royal jelly.

We have previously reported that royal jelly (RJ) from honeybees (Apis mellifera) has weak estrogenic activity mediated by interaction with estrogen receptors that leads to changes in gene expression and cell proliferation. In this study, we isolated four compounds from RJ that exhibit estrogenic activity as evaluated by a ligand-binding assay for the estrogen receptor (ER) beta. These compounds were identified as 10-hydroxy-trans-2-decenoic acid, 10-hydroxydecanoic acid, trans-2-decenoic acid and 24-methylenecholesterol. All these compounds inhibited binding of 17beta-estradiol to ERbeta, although more weakly than diethylstilbestrol or phytoestrogens. However, these compounds had little or no effect on the binding of 17beta-estradiol to ERalpha. Expression assays suggested that these compounds activated ER, as evidenced by enhanced transcription of a reporter gene containing an estrogen-responsive element. Treatment of MCF-7 cells with these compounds enhanced their proliferation, but concomitant treatment with tamoxifen blocked this effect. Exposure of immature rats to these compounds by subcutaneous injection induced mild hypertrophy of the luminal epithelium of the uterus, but was not associated with an increase in uterine weight. These findings provide evidence that these compounds contribute to the estrogenic effect of RJ.

Royal jelly stimulates bone formation: physiologic and nutrigenomic studies with mice and cell lines.

Royal jelly (RJ) has diverse physiological and pharmacological functions. We observed its weak estrogenic activity in the previous study. RJ stimulated the proliferation of mouse osteoblast-like MC3T3-E1 cells at 0.1 mg/ml, and the effect was blocked by the specific estrogen receptor antagonist ICI 182,780. The addition of 0.1-1.0 mg/ml RJ enhanced collagen production in culture medium. Oral administration of RJ to normal female mice for 9 weeks increased the ash content of their tibiae. DNA microarray analysis revealed significant changes in gene expression related to extracellular matrix formation when the femurs of mice fed RJ were analyzed. Quantitative reverse transcriptase-PCR (RT-PCR) confirmed up-regulation of procollagen I alpha1 gene expression. These data suggest that RJ as a whole or some of its individual components stimulates production of type I collagen and other activities for bone formation through action on osteoblasts.

Role of sevelamer hydrochloride in the coronary artery calcification score.

The coronary artery calcification score (CACS) is higher in hemodialysis (HD) patients than in non-HD patients for each age group from the fifth to the eighth decade of life. In order to clarify the relationship between the rate of change in the CACS and several factors related to calcium (Ca) and phosphate (P) metabolism in HD patients, we determined the CACS twice in 144 HD outpatients at an interval of approximately 12 months (2003 and 2004). The dosage of vitamin D formulations (alfacalcidol or maxacalcitol) was reduced or ceased if the serum Ca concentration exceeded 5.0 mEq/L, or the serum P concentration exceeded 6.0 mg/dL, and the dosage of combined sevelamer hydrochloride (SH) and calcium carbonate (CaCO3), as the phosphate binder, was adjusted to maintain the concentrations below these levels. The study parameters were: (1) the total dosage of alfacalcidol (microg), maxacalcitol (microg), SH (mg), and CaCO3 at the time of each CACS measurement; and (2) serum concentrations of Ca, P, alkaline phosphatase, high-sensitivity parathyroid hormone (HS-PTH), total protein, albumin, total cholesterol, triglycerides (TG). Regression analysis showed a significant correlation among the total SH dosage, TG, and alphaCACS. Future investigations will include the differences in alphaCACS between patients treated with SH who experience a rise in Ca and/or P and those with a decrease in Ca and/or P.

Identification of caffeoylquinic acid derivatives from Brazilian propolis as constituents involved in induction of granulocytic differentiation of HL-60 cells.

We have previously reported that Brazilian propolis extracts inhibited growth of HL-60 human myeloid leukemia cells, which is partly attributed to the induction of apoptosis associated with granulocytic differentiation. In this study, we isolated three compounds which induce granulocytic differentiation evaluated by nitroblue tetrazolium (NBT)-reducing assays from the water extract of propolis and identified as 4,5-di-O-caffeoylquinic, 3,5-di-O-caffeoylquinic, and 3,4-di-O-caffeoylquinic acids by NMR analysis. Cell growth inhibitory activity of these caffeoylquinic acids was found in HL-60 cell, which was mainly attributed to the induction of apoptosis. Furthermore, the potency of caffeoylquinic acid derivatives to induce granulocytic differentiation was examined in HL-60 cells. Caffeic, quinic, and chlorogenic acids had no effects on the NBT-reducing activity, while 3,4,5-tri-O-caffeoylquinic acid induced more than 30% of NBT-positive cells. These results suggest that the number of the caffeoyl groups bound to quinic acid plays an important role in the potency of the caffeoylquinic acid derivatives to induce granulocytic differentiation. This is the first report demonstrating that the caffeoylquinic acid derivatives induce granulocytic differentiation of HL-60 cells.

Effects of propolis on cell growth and gene expression in HL-60 cells.

Brazilian propolis obtained from honeybee hives was extracted with water or ethanol. Cell growth-inhibitory activities of these propolis extracts were found in HL-60 human myeloid leukemia cells. The extracts-induced apoptosis in the cells, which was characterized by morphological and nucleosomal DNA fragmentation analysis. The apoptosis was mainly attributed to the induction of granulocytic differentiation, which was evaluated by nitro blue tetrazolium (NBT) reducing assays and cytofluorometric analysis for the expression of cell surface marker CD11b. DNA microarray analysis was performed to examine the gene expression profiles in the propolis-treated HL-60 cells accompanied with granulocytic differentiation, which were compared with those in all-trans retinoic acid-treated cells. Several genes were up- or down-regulated. Two genes encoding S100 calcium binding protein A9 and ferritin, heavy polypeptide 1 were up-regulated, which were also confirmed by semi-quantitative reverse transcriptase-PCR (RT-PCR). Propolis-induced growth inhibition in HL-60 cells was, at least in part, due to differentiation with gene expression profiles, which are similar to those induced by all-trans retinoic acid.

Osmotic stress in barley regulates expression of a different set of genes than salt stress does.

Under high salt conditions, plant growth is severely inhibited due to both osmotic and ionic stresses. In an effort to dissect genes and pathways that respond to changes in osmotic potential under salt stress, the expression patterns were compared of 460 non-redundant salt-responsive genes in barley during the initial phase under osmotic versus salt stress using cDNA microarrays with northern blot and real-time RT-PCR analyses. Out of 52 genes that were differentially expressed under osmotic stress, 11, such as the up-regulated genes for pyrroline-5-carboxylate synthetase, betaine aldehyde dehydrogenase 2, plasma membrane protein 3, and the down-regulated genes for water channel 2, heat shock protein 70, and phospholipase C, were regulated in a virtually identical manner under salt stress. These genes were involved in a wide range of metabolic and signalling pathways suggesting that, during the initial phase under salt stress, several of the cellular responses are mediated by changes in osmotic potential.