Antler Extract Research Articles

Antioxidant, Anti-acetylcholinesterase and Composition of Biochemical Components of Russian Deer Velvet Antler Extracts

Russian deer velvet antlers were divided into three parts and subjected to a extraction process using hot water at 100, 110, and <TEX>$120^{\circ}C$</TEX> or an extraction with 70% ethanol. Each extract was analyzed for its biochemical components, including uronic acid, sulfated-glycosaminoglycans (sulfated-GAGs), and sialic acid, and the antioxidant and anti-acetylcholinesterase activities were investigated. Different levels of uronic acid and sulfated-GAGs were observed in the extracts according to the water temperature used for the extraction, and contents decreased with increasing extraction temperature. The upper layer of each extract showed high amounts of uronic acid and sulfated-GAGs, followed by the middle and base layers. Ethanol extraction was more effective for recovering uronic acid than sulfated-GAGs. Sialic acid content was the highest in the <TEX>$110^{\circ}C$</TEX> extracts but was not observed in the ethanol extracts. Velvet antler extracts showed strong antioxidant activities against DPPH and hydrogen peroxide as well as strong reducing power in a dose-dependent manner. However, the antioxidant activities were different in each layer and according to the extraction method. Additionally, velvet antler extracts exhibited inhibitory activity against acetylcholinesterase, which is associated with Alzheimer's disease, in a dose-dependent manner. These results suggest that velvet antler extracts are useful as a functional food ingredient and/or a pharmaceutical.

A study on the extracting characteristics of velvet antlers using kyenegum protease

ABSTRACT Objective：Kyenegum has been frequently used for characterizing digestive symptoms in the traditional and oriental medicines. This study was conducted to investigate the characteristics of extracts from velvet antlers using the 4 different kinds of extracting methods.Methods：The extracts of velvet antlers were extracted using a 65℃ DW (9hrs), a Kyenegum crude enzyme, a 121℃ DW (2hrs), and a Kyenegum protease. To evaluate the characteristic of velvet antler extracts, we examined the brix, soluble solid, amino acid, mineral composition, and collagen protein.Results：As a result of the comparisons of velvet antlers extracted by the traditional extraction and the crude enzyme of kyenegum, the brix and soluble solid showed the higher contents for kyenegum enzymes. Also, mineral contents of the extracted velvet antlers were higher, p articularly in Ca and P for those. The contents of collagen protein, hydroxyproline and hydroxylysine, were fou nd to be more than twice in kyenegum protease compared with other extracting methods.Conclusion：These results indicated that the Kyenegum crude enzyme and protease are very effective to extract of velvet antlers.Key words：Kyenegum, velvet antlers, amino acid, collagen protein, mineral

Effect of aqueous antler extract on scopolamine-induced memory impairment in mice and antioxidant activities

This study was performed to estimate ameliorating effect of aqueous antler extract (AAE) on memory impairments induced by scopolamine (SCOP, 2 mg/kg, i.p.). Tacrine (THA, 10 mg/kg, p.o.) was used as a positive control. The passive avoidance test and Y-maze test results showed that a single AAE (200 mg/kg, p.o.) administration significantly restored memory impairment. Moreover, AAE or THA treatment significantly reduced the escape latency prolonged by SCOP during trial sessions in the Morris water maze test. In vivo study, AAE significantly inhibited acetylcholinesterase (AChE) activity (p<0.001), whereas choline acetyltransferase (ChAT) activity (p<0.05) was enhanced. In addition, SCOP elevated oxidative damage in the brain. However, treatment with AAE to the amnesic mice induced by SCOP considerably decreased malondialdehyde (MDA) level (p<0.01) and restored the activities of superoxide dismutase (SOD) (p<0.01) and glutathione peroxidase (GSH-Px) (p<0.001) approaching the control values. These results suggest that AAE showed the potential cognitive-enhancing activity by regulating cholinergic marker enzyme activities and promoting the antioxidant system.

Composition of Biologically Active Substances and Antioxidant Activity of New Zealand Deer Velvet Antler Extracts

Deer velvet antler was subjected to the extraction process using boiling water at three different temperatures (100, 110 and <TEX>$120^{\circ}C$</TEX>) and 70% ethanol solution. Functional components such as uronic acid, sulfated-glycosaminoglycans (sulfated-GAGs) and sialic acid in the extracts were analyzed, and their antioxidant activities were investigated using several in vitro models. Uronic acid and sulfated-GAGs content of each extract significantly decreased with increasing extraction temperature (p<0.05), while the residues obtained from the upper and middle part of the antler had a higher uronic acid content than the residues obtained from the base section. Sialic acid contents were highest in compounds extracted at <TEX>$110^{\circ}C$</TEX>, followed by 120 and <TEX>$100^{\circ}C$</TEX>. The 70% ethanol extracts also had a high levels of uronic acid content, but not for sulfated-GAGs and sialic acid. All extracts showed good antioxidant ability in a dose-dependant manner, with the <TEX>$100^{\circ}C$</TEX> residue exhibiting the strongest activity compared to the 110 and <TEX>$120^{\circ}C$</TEX> extracts. In relation to the hydroxyl radical scavenging activity and reduction power, the 70% ethanol extract exhibited the strongest activity. Furthermore, the velvet antler extracts inhibited apoptosis in hydrogen peroxide-induced PC-12 cells.

Stimulation of the Extracellular Matrix Production in Dermal Fibroblasts by Velvet Antler Extract

Fibroblasts produce many components of the extracellular matrix (ECM) and so they contribute to the maintenance of connective tissue integrity. The aim of this study is to evaluate the effect of velvet antler extract (VAE) on the ECM production of dermal fibroblasts cultured in vitro. Primary cultured human dermal fibroblasts were treated with VAE, and then the ECM production was determined by RT-PCR, ELISA and Western blot analysis. Furthermore, the change of gene expression according to VAE treatment was evaluated by cDNA microarray. VAE accelerated the growth of fibroblasts in a dose-dependent manner. VAE increased the production of several ECM components, including type 1 collagen, fibronectin and elastin. In line with these results, the phosphorylations of p42/44 ERK and p38 mitogen-activated protein kinase were markedly increased by VAE, suggesting that the enhancement of ECM production may be linked to the activation of intracellular signaling cascades. VAE also significantly increased cell migration on an in vitro scratch wound test. In cDNA microarray, many genes related with connective tissue integrity were identified to be up-regulated by VAE. These results suggest that VAE has a potential to stimulate ECM production, and VAE may be applicable for maintaining the skin's texture.

Phenylhydrazine으로 유도한 용혈성 빈혈 흰쥐에 대한 녹용 추출물의 조혈 효과

본 연구는 부위별 녹용 추출물이 PHZ 투여로 용혈성 빈혈을 유도한 흰쥐의 조혈작용에 대한 효과를 검증하고자 하였다. 4주령의 암컷 Sprague-Dawley 흰쥐에 PHZ 10 ㎎/㎏을 4일간 미정맥 주사하여 용혈성 빈혈을 유발한 후 각 부위별 녹용 추출물 200 ㎎/㎏을 일주일간 경구투여 하였다. 용혈성 빈혈 유발군인 PHZ군은 hemoglobin, hematocrit치, 적혈구수가 대조군에 비해 유의적으로 감소하였고, 망상적혈구수는 유의적으로 증가하였다. 반면, 녹용 추출물 군에서는 hemoglobin, hematocrit치, 적혈구수가 PHZ 투여군보다 증가하였다. 특히 분골, 상대, 중대 추출물 투여 시 유의성 있게 증가하였다. 상대 추출물 투여군에서는 망상적혈구수 증가가 유의적으로 억제되었다. PHZ 투여로 증가된 혈청 EPO 함량은 녹용 추출물 투여군에서 모두 정상적으로 회복되었다. Hemoglobin 합성에 관여하는 δ-ALAD 활성은 녹용 추출물 투여군에서 활성이 증가되는 결과를 보여주었다. 따라서 녹용 추출물은 PHZ으로 유도한 용혈성 빈혈에 대한 바람직한 조혈효과가 있는 기능성 물질로 사료된다.

Effect of antler extract on corticosteroid-induced avascular necrosis of the femoral head in rats

Ethnopharmacological relevance Deer antler, traditionally used as a tonic and valuable drug in oriental medicine, has been considered to possess bone-strengthening activity and effectively used in bone diseases therapy. Aim of the study The present study was designed to investigate therapeutic effect of antler extract on avascular necrosis of the femoral head (ANFH) induced by corticosteroids in rats. Materials and methods Rats were intragluteally injected with dexamethasone at 50 mg/kg twice per week for 6 weeks to induce ANFH. Then the rats were treated with antler extract by oral gavage at 200 mg/kg, 400 mg/kg and 800 mg/kg once per day for 60 days. The concentration of hydroxyproline and hexosamine in serum was measured and the ultrastructure of femoral head was examined. In vitro, effect of the drug-containing serum of antler extract on proliferation and differentiation of primary osteoblasts were investigated by MIT assay, ALP activity assay and cell cycle analysis. Results After treatment with antler extract, the degree of necrosis induced by dexamethasone was significantly reduced, hydroxyproline was significantly decreased, and hexosamine and the ratio of hexosamine/hydroxyproline were significantly increased. The drug-containing serum of antler extract promotes osteoblastic proliferation through regulation of cell cycle progression. Conclusions The results suggest that antler extract has a positive curative effect on ANFH by promoting osteoblastic proliferation.

발효녹용의 균주 선별 및 발효녹용의 생리활성

This study was conducted to isolate strain for the preparation of fermented antler (Cervus cornu parvum) and evaluate its physiological activities. The growth degrees of twenty-one samples from Bacillus sp., Lactobacillius sp. and mushroom strain on antler extract agar were evaluated in this study, and Bacillus subtilis KH-15, SCB-3, Cordyceps militaris, Phellinus linteus, Inonotus obliquus 26136, and Inonotus obliquus 26147 were selected. The fermented antler extract by C. militaris had relatively higher contents of total sugar (1619.3 /mL), uronic acid (302.0 /mL), sulfated-glycosaminoglycan (S-GAGs) (119.9 /mL) and sialic acid (21.6 /mL) than any other extracts. The anti-complementary activities of all fermented antler extracts were higher than non-fermented antler extract, and among these samples, fermented antler extract by C. militaris showed the highest anti-complementary activity (inhibition of 50% total complement hemolysis, ; 50.1% at 1,000 /mL). The ability of fermented antler extract by B. subtilis KH-15 to scavenge 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical (; 4.97 mg/mL) was significantly the highest (p; 16.98 mg/mL) among all samples. The results of this study suggest that physiological effects including immuno-modulating and antioxidant activities of the antler may be increased through fermentation process.

녹용 추출물이 치매 동물모델의 기억력 개선과 관련효소 활성에 미치는 효과

The aim of this study was to investigate the ameliorating effects of deer antler extract on the learning and memory impairments induced by the administration of scopolamine (2 ㎎/㎏, i.p.) in rats. Tacrine was used as a positive control agent for evaluating the cognition enhancing activity of deer antler extract in scopolamine- induced amnesia models. The results showed that the deer antler extract-treated group (200 ㎎/㎏, p.o.) and the tacrine-treated group (10 ㎎/㎏, p.o.) significantly ameliorated scopolamine-induced amnesia based on the Morris water maze test. Although there was no statistical significance of brain ACh contents among the experimental groups, the brain ACh contents of the deer antler extract-treated group was slightly higher than that of the scopolamine-treated group. The inhibitory effect of deer antler extract on the acetylcholinesterase activity in the brain was significantly lower than that of scopolamine-treated group. The tacrine- and the deer antler-treated groups reduced the MAO-B activity compared to the scopolamine-treated group, but not significantly. These results suggest that the deer antler extract could be an effective agent for the prevention of the cognitive impairment induced by cholinergic dysfunction.

Effect of Cervus korean TEMMINCK var. mantchuricus Swinhoe on protease activities, antioxidant and free radical damages in rheumatis arthritis rats

The effect of deer antler extracts (DAA) of Cervus korean TEMMINCK var. mantchuricus Swinhoe on protease activities, oxidant and free radical damages in synovial fluid from rheumatoid arthritis in rats was studied. Rats were i.p. administered with DAA. We have compared (using the same series of experimental samples) the levels of activity of a comprehensive range of cytoplasmic, lysosomal and matrix protease types, together with the levels of free radical induced protein damage (determined as protein carbonyl derivative) and total antioxidant in synovial fluid from rheumatoid arthritis (RA) and DAA-treated rats. Many proteases activities were shown to be significantly increased in RA compared to normal rats. Protease activities (including those enzyme types putatively involved in the immune response, such as dipeptidyl aminopeptidase IV) in plasma were not significantly different between RA and normal rats. DAA treatment at dose of 100 μg/kg suppressed the production of the proteases of cytoplasmic, lysosomal and matrix protease types. The level of free radical induced damage to synovial fluid proteins was approximately 2-fold lower in DAA rats compared to RA rats, although there was no significant difference in total antioxidant status in synovial fluid or plasma between RA and DAA rats. It was concluded that DAA treatment reduces the activation of proteolytic enzymes and free radicals, which are likely to be of equal potential importance as protein damaging agents in the pathogenesis of RA.

Chloroform extract of deer antler inhibits osteoclast differentiation and bone resorption

It has been reported that deer antler extract has anti-bone resorptive activity in vivo. However, little is known about the cellular and molecular mechanism of this effect. In this study, we investigated the effects of deer antler extracts on osteoclast differentiation and bone-resorption in vitro. Chloroform extract (CE-C) of deer antler inhibited osteoclast differentiation in mouse bone marrow cultures stimulated by receptor activator of NF-κB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF). CE-C suppressed the activation of extracellular signal-regulated kinase (ERK), protein kinase B (PKB/Akt) and inhibitor of kappa B (I-κB) by RANKL in osteoclast precursor cells. It also inhibited the bone resorptive activity of differentiated osteoclasts that was accompanied by disruption of actin rings and induction of the apoptosis. These results demonstrate deer antler extract may be a useful remedy for the treatment of bone-resorption diseases such as osteoporosis.

A deer antler extract prevents bone loss in postmenopausal osteoporosis model rats

This study was conducted to examine the effects of a deer antler extract in water (DAE) on bone metabolism. 8 week-old female rats (Sprague-Dawley) were subjected to establish the postmenopausal osteoporosis model by feeding a low calcium diet for 4 weeks after ovariectomy operation. The model rats were divided into 4 groups (n=32); Control, without DAE DAE 2.5, treated with 2.5% (wt/wt) DAE 5.0, treated with 5% (wt/wt) Est, treated with 17β-estradiol (10μg/kg BW). The rats were fed basal diet (AIN-93M) or experimental diets containing two levels of DAE for 6 weeks. Serum, femur and urine samples were collected. Weight gain was not affected by DAE supplementations. Femur weight and mineral contents (Ca, Mg) were significantly increased both by DAE supplementations and estrogen treatment compared to control. Serum estradiol levels were significantly elevated in DAE-fed groups than in the control group. In both DAE-fed and Est-treated groups, urinary hydroxyproline (OHPr) and deoxypyridinoline (DPD) excretion were significantly decreased in contrast to control group. Also serum interleukin-6 (IL-6) decreased both by DAE supplementations and Est treatment, suggesting that bone-sparing effect could be partly attributed to the modulation of osteoclastogenesis induced by IL-6. These results indicate that a DAE has beneficial effects on estrogen-dependent bone loss and can be considered as an alternative functional substance for estrogen replacement therapy (ERT) in treating postmenopausal osteoporosis.

IMMUNOSUPPRESSIVE ACTIVITY OF DEER ANTLER EXTRACTS OF CERVUS KOREAN TEMMINCK VAR. MANTCHURICUS SWINHOE, ON TYPE II COLLAGEN-INDUCED ARTHRITIS

Unossified horn or pilose antler cut from deer, which belong to the Cervidae generally is termed Nokyong. Nokyong is one of the most famous Korean traditional medicines and has been considered to possess sexual-reinforcing and antiaging actions. In this study, water extract of deer antler extract (DAA) prepared from the growing antler of Cervus korean TEMMINCK var. mantchuricus Swinhoe was used to investigate the efficacy of the DAA on the development of type II collagen (CII)-induced arthritis (CIA) in rats. Male rats were immunized with an emulsion of 200 microg of CII and complete Freund's adjuvant (CFA). The rats then were administered by injection a suspension of DAA or phosphate-buffered saline. The effect of DAA on cellular responses to CII was examined. The injection of DAA suppressed the CII-specific secretion of interferon (IFN)-gamma from splenocytes ex vivo. The influence of DAA also was evaluated on the incidence and development of arthritis in rat CIA. Rats were immunized twice at a 3-wk interval with bovine CII, with DAA being given by injection once a d for 14 d with four different regimens. A 14-d course of DAA treatment at a daily dose of 100 microg/kg, which began on the d of the first CII immunization, suppressed the development of arthritis, as well as antibody formation and delayed-type hypersensitivity to CII. Treatment with DAA resulted in inhibition of development of arthritis and immune responses to CII.

Studies on the Effects of Biomedicinal Agents on Serum Concentration of Ca2+, P and ALP Activity in Osteoporosis-Induced Rats

The present study was carried out to investigate the effects of biomedicinal agents on Ca2+, P and alkaline phosphatase (ALP) levels in ovariectomized rats. Rats were ovariectomized bilaterally and were fed up with Ca2+ and P-free diet during 8(9,10) weeks to induce osteoporosis. Osteoporosis was determined by the extent of bone density and by lowering the concentrations of serum Ca2+, P and ALP activity every week. Rats in antler, safflower, ipriflavon, or co-administrated with estrogen groups were administrated with feed supplement for 5 weeks to elucidate the protective and therapeutic effects against osteoporosis. The bone tissue was examined with electron microscope to determine the effects of each treatment on osteoporosis. 1. The levels of serum Ca2+ and P in osteoporosis-induced rats, administrated with antler, ipriflavon and estrogen groups, were little higher than those of control rats. However, the levels of serum Ca and P in ovariectomized rats were significantly higher than those of control group (p<0.05). 2. The activities of serum ALP in osteoporosis-induced rats, administrated with antler extract, safflower, ipriflavon, or co-administrated with estrogen, were little increased in comparing with those of control group, but were significantly decreased in with combination of estrogen for 5 weeks. However, The connections were interrupted and the bone matrix was destroyed in the osteoporosis-induced rats. 3. The inter-trabecular connections were examined under electron microscope. The connections were well maintained and bone loss was without in the administration with antler, safflower, and ipriflavon with combination of estrogen for 5 weeks. However, The connections were interrupted and the bone matrix was destroyed in the osteoporosis-induced rats.

Lysophosphatidylcholine derived from deer antler extract suppresses hyphal transition in Candida albicans through MAP kinase pathway

A family of 2-lysophosphatidylcholines (lyso-PCs) was isolated from deer antler extract, guided exclusively by hyphal transition inhibitory activity in Candida albicans. Structural determination of the isolated lyso-PCs by spectroscopic methods, including infrared spectroscopy, 1H nuclear magnetic resonance (NMR), 13C NMR, 2D correlation spectroscopy NMR, fast atom bombardment mass spectrometry and tandem mass spectrometry, confirmed that the natural products were composed of at least four different lyso-PCs varying in fatty acid moiety at the sn-1 position of the glycerol backbone. The major lyso-PCs were confirmed as 1-stearoyl-, 1-oleoyl-, 1-linoleoyl- and 1-palmitoyl-2-lyso- sn-glycero-3-phosphatidylcholines. Lyso-PC specifically suppressed the morphogenic transition from yeast to hyphae in C. albicans, without affecting the growth of either yeast or hyphae. Lyso-PC exerted hyphal transition that suppressed activity in the broad spectrum of the Candida species, such as C. albicans, Candida krusei, Candida guilliermondii and Candida parapsilosis. Northern analysis indicated that the suppression was mediated through the mitogen-activated protein kinase pathway.

Effect of water-soluble extract from antler of wapiti (Cervus elaphus) on the growth of fibroblasts

The water-soluble extracts were prepared from the tip sections of antlers of 4-yr-old wapiti stags, and the effect of extract on the growth of bovine skin fibroblasts in culture was examined. The results showed the presence of growth promoting factor(s) in the antler extract. The stimulation of cell growth was found to be dose dependent (P &lt; 0.05). Key words: Growth factor, antler, wapiti, fibroblast

A new monitoring system of cultured myocardial cell motion: Effect of pilose antler extract and cardioactive agents on spontaneous beating of myocardial cell sheets.

Effects of various cardioactive agents and a water extract of the pilose antler of Cervus nippon var. mantchuricus on periodic beating of cultured myocardial cell sheets were examined by using an image analyzing system. Norepinephrine increased the beating rate and the beating amplitude, whereas digoxin and forskolin enlarged only the beating amplitude. Verapamil and propranolol decreased both the beating rate and the beating amplitude. The water extract of the pilose antler showed no remarkable effects in a standard medium (2.1 mM Ca2+). However, it significantly increased the beating amplitude when the beating was suppressed by replacement with a low calcium medium (0.5 mM Ca2+). A similar effect was found for 70% ethanol-soluble and -insoluble fractions of the extract.

Effects of pilose antler extracts on doxorubicin‐induced beating abnormalities of cultured myocardial cells

AbstractEffects of pilose antler extracts and subsequent fractions on normal and doxorubicin (DX)‐administered cultured myocardial cells from mouse embryo were investigated. A water extract did not affect the percentage of beating cells at 1–100 μg/mL but suppressed the beating rate at 100 μ/mL. A pretreatment with the water extract significantly restored the percentage of beating cells which was reduced by administration of DX, and showed a tendency to recover the reduced beating rate. A similar pretreatment with an aqueous EtOH‐solnble portion (fraction 1) of the water extract significantly recovered both the percentage of beating cells and the beating rate, but further fractionation did not improve the activities. LDH leakage from DX‐administered myocardial cells was significantly diminished by the pretreatment with fraction 1, though ATP content in the cells was not appreciably recovered.

Effects of repeated administration of deer antler extract on biochemical changes related to aging in senescence-accelerated mice.

Several biochemical parameters related to aging were evaluated after a hot-water extract prepared from unossified pilose antler of Cervus nippon TEMMINCK var. mantchuricus Swinhoe (Rokujo) had been administered orally for 8 successive days to senescence-accelerated mice (SAM), a novel murine model of spontaneously promoted aging. It was found that the repeated oral administration of Rokujo had significant restoring effects on the physiological degenerations which were associated with the development of senile symptoms. These effects included (1) an increase in the plasma testosterone level in male mice; (2) a decrease in the contents of malondialdehyde in the liver and brain; (3) an increase in the liver protein contents; (4) an increase in the liver superoxide dismutase activity; and (5) a decrease in the activity of monoamine oxidase B in the liver and brain membranes. Most of these effects were selectively observed in the senile-prone strain of SAM (SAM-P) as compared to the normal, resistant strain (SAM-R). The present data suggest that Rokujo may exert an anti-aging action in male senile animals.

Stimulating effect of deer antler extract on protein synthesis in senescence-accelerated mice in vivo.

The effects of subchronic administration of Rokujo (Cervus nippon TEMMINCK var. mantchuricus Swinhoe) extract on protein, ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) syntheses in vivo were investigated in male senescence-accelerated mice (SAM). At 20 min after intraperioneal injection of amino acid and nucleosides labeled with radioisotpes, the incorporation of [14C]leucine into the serum protein was increased in the Rokujo-treated mice, in a parallel manner to the elevation of the serum protein content. Increased incorporation of [14C]uridine into RNA as well as that of [14C]leucine into protein was found in the liver and kidney of the Rokujo-treated mice. Further experiments revealed that although a brief exposure to Rokujo extract in vitro did not directly affect the RNA polymerase activity, the activity of RNA polymerase II in solubilized liver nuclei of Rokujo-treated mice was markedly accelerated. These results suggest that enhancement of RNA polymerase II activity induced by long-term Rokujo treatment is responsible for the increase in protein synthesis in vivo in SAM.