Twenty-four-month-old male C57BU6 mice with low serum testosterone levels were used as a late-onset hypogonadism （LOH） animal model for examining the effects of velvet antler polypeptide （VAP） on sexual function a...Twenty-four-month-old male C57BU6 mice with low serum testosterone levels were used as a late-onset hypogonadism （LOH） animal model for examining the effects of velvet antler polypeptide （VAP） on sexual function and testosterone synthesis. These mice received VAP for 5 consecutive weeks by daily gavage at doses of 100, 200, or 300 mg kg-1 body weight per day （n = 10 mice per dose）. Control animals （n = 10） received the same weight-based volume of vehicle. Sexual behavior and testosterone levels in serum and interstitial tissue of testis were measured after the last administration of VAP. Furthermore, to investigate the mechanisms of how VAP affects sexual behavior and testosterone synthesis in vivo, the expression of steroidogenic acute regulatory protein （STAR）, cytochrome P450 cholesterol side-chain cleavage enzyme （P450scc）, and 3β-hydroxysteroid dehydrogenase （3βHSD） in Leydig cells was also measured by immunofluorescence staining and quantitative real-time PCR. As a result, VAP produced a significant improvement in the sexual function of these aging male mice. Serum testosterone level and intratesticular testosterone （ITT） concentration also increased in the VAP-treated groups. The expression of STAR, P450scc, and 3β-HSD was also found to be enhanced in the VAP-treated groups compared with the control group. Our results suggested that VAP was effective in improving sexual function in aging male mice. The effect of velvet antler on sexual function was due to the increased expression of several rate-limiting enzymes of testosterone synthesis （STAR, P450scc, and 3β-HSD） and the following promotion of testosterone syothesis in vivo.展开更多
Background Velvet antler polypeptides (VAPs), which are derived from the antler velvets, have been reported to maintain survival and promote growth and differentiation of neural cells and, especially the development ...Background Velvet antler polypeptides (VAPs), which are derived from the antler velvets, have been reported to maintain survival and promote growth and differentiation of neural cells and, especially the development of neural tissues This study was designed to explore the influence of VAPs on neural stem cells in vitro derived from embryonic rat brain Methods Neural stem cells derived from E12 14 rat brain were isolated, cultured, and expanded for 7 days until neural stem cell aggregations and neurospheres were generated The neurospheres were cultured under the condition of different concentration of VAPs followed by immunocytochemistry to detect the differentiation of neural stem cells Results VAPs could remarkablely promote differentiation of neural stem cells and most neural stem cells were induced to differentiate towards the direction of neurons under certain concentration of VAPs Conclusion Neural stem cells can be successfully induced into neurons by VAPs in vitro , which could provide a basis for regeneration of the nervous system展开更多
A novel polypeptide having stimulant effect on some cell proliferation was isolated from the velvet antler (Cervus elaphus Linnaeus). The velvet antler polypeptide consists of a single chain of 32 amino acid residues...A novel polypeptide having stimulant effect on some cell proliferation was isolated from the velvet antler (Cervus elaphus Linnaeus). The velvet antler polypeptide consists of a single chain of 32 amino acid residues. Amino acid sequence of the polypeptide was identified as: VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM.展开更多
天然的丝状真菌是我国重要的微生物资源,能产生大量有益的初生、次生代谢产物,其代谢受到各类调控因子的调控。天鹅绒(Velvet)蛋白家族是真菌中最独特且高度保守的一类特殊调控蛋白,可调控真菌的生长、分化和次生代谢物的生物合成。该...天然的丝状真菌是我国重要的微生物资源,能产生大量有益的初生、次生代谢产物,其代谢受到各类调控因子的调控。天鹅绒(Velvet)蛋白家族是真菌中最独特且高度保守的一类特殊调控蛋白,可调控真菌的生长、分化和次生代谢物的生物合成。该文结合国内外的研究进展综述Velvet蛋白家族Velvet A(VeA)、Velvet like B(VelB)在真菌中的结构及其蛋白复合物对生长发育和次生代谢的调控作用,可为进一步探索Velvet蛋白家族调控真菌次生代谢物的生物合成通路、激活沉默基因簇、挖掘新的化合物提供参考。展开更多
Supercritical CO2 (SC-CO2) extraction technology and ultrasonic technology were used to extract two active sex hormones, estradiol and progesterone, and insulin-like growth factor-1 (IGF-1) from antler velvet. The eff...Supercritical CO2 (SC-CO2) extraction technology and ultrasonic technology were used to extract two active sex hormones, estradiol and progesterone, and insulin-like growth factor-1 (IGF-1) from antler velvet. The effects of SC-CO2 extraction condition on the extraction yield and content of sex hormones, the ultrasonic extrac-tion condition on the content of IGF-1 and the SC-CO2 extraction condition on the activity remaining of IGF-1 were studied. The optimal conditions were obtained. The experimental results showed that, in presence of 75% ethanol as the co-solvent, the mean yield and content of estradiol and progesterone were 87.67 pg·g-1 and 1224.10 pg·g-1, 12.38 ng·g-1 and 354.06 ng·g-1, respectively, with extraction pressure of 30 MPa, temperature of 35°C, extraction time of 30 min and CO2 consumption of 15 L·g-1 at the flow rate of 2.0 L·min-1. The highest content of IGF-1 was 7425.75 ng·g-1 antler velvet residue, when the pH10 ammonia-ammonium chloride buffer solution was used as the solvent, the ratio of solvent to sample was 20/1 (volume/mass), the extraction temperature was 0-35°C, and the ex-traction time was 4×15 min. Under these conditions, 93.68% activity remaining of IGF-1 in the residue was ob-tained, while little IGF-1 activity exists in traditional residue. The experimental results indicate that the technology of SC-CO2 with co-solvent is of advantage for getting high content sexual hormones and keeping high activity of IGF-1 in the residue, which can not be achieved by traditional extraction methods.展开更多
Biochemical techniques including ion-exchange chromatography, gel filtration chromatography and re- versed-phase high-performance liquid chromatography（RP-HPLC） were used to isolate and purify the natural poly- pept...Biochemical techniques including ion-exchange chromatography, gel filtration chromatography and re- versed-phase high-performance liquid chromatography（RP-HPLC） were used to isolate and purify the natural poly- peptide from velvet antler（nVAP） of Cervus elaphus（C, elaphus）, which has a molecular weight of 3215.8 and the primary structure of VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM. The homology of the protein sequence in nVAP with known protein sequence is less than 50%, suggesting that nVAP appears to be a new bioactive substance. At a level of 0.4--50 gg/mL, nVAP promotes mitosis in epidermal ceils, chondrocytes and NIH3T3 fibroblasts pri- marily cultured in a significant way. Given that a yield of high-purity nVAP isolated from C. elaphus is 0.001%, nVAP is artificially synthesized to prepare synthetic velvet antler polypeptide（sVAP） according to its primary struc- ture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） of sVAP shows a single band, and its HPLC spectrum displays a single peak. The matrix-assisted laser desorption ionization time-of-flight mass spectro- metry（MALDI-TOF-MS） was used to identify sVAP to be of a molecular weight of 3200 and the consistency between primary structures of sVAP and nVAE Bioactivity test shows that at a dose of 5--40 μg/mL, sVAP promotes the pro- liferation of primarily cultured epidermal cells and NIH3T3 cell line. From the traditional Chinese medicine theory, velvet antler from Cervus nippon（C, nippon） and velvet antler from C. elaphus are considered as the same medicine, but differences between biochemical base and pharmacological effect of these two velvet antlers have been observed. We compared the total polypeptide mapping of the two velvet antlers, discovering that nVAP is active polypeptide and only exists in the velvet antler of C. elaphus, sVAP is similar to nVAP in physicochemical property and biological activity. These studies extend the possible utility of sVAP to be the promising compound to prepare velvet antler pol展开更多
Acer velutinum Boiss is a valuable tree species native to Iran, and its seeds possess physiological dormancy that hampers seedling production in the nursery for large-scale reforestation efforts. The aim of this study...Acer velutinum Boiss is a valuable tree species native to Iran, and its seeds possess physiological dormancy that hampers seedling production in the nursery for large-scale reforestation efforts. The aim of this study was to determine the optimal dormancy breaking treatments for A. velutinum seeds. We conducted a factorial experiment involving six seed lots collected along an elevation gradient from 300 to 1800 m at 300 m interval and four cold-moist stratification periods （0, 4, 8 and 16 weeks） at 4℃ and 70% relative humidity. The result shows that the germination of cold-moist stratified seeds was significantly （p 〈 0.0001） higher than the control for all seed lots. The highest germination capacity was recorded after 16 weeks of cold-moist stratification for all seed lots （68% 88% depending on the seed lot） except those collected from mid altitude sites （600 and 900 m） that germinated equally well （≥ 75%） after 4and 8-week of clod-moist stratification compared to the other seed lots. The mean germination time was significantly shorter （12 to 19 days, depending on the seed lot） for seeds stratified for 16 weeks than for untreated seeds. It can be concluded that： （1） cold-moist stratification for 16 weeks is the best pre-sowing treatment for breaking dormancy in A. velutinum seeds; and （2） seeds should be collected from mid altitude sites （600 and 900 m） to get more than 80% germination within 15 days, and these seed lots even required shorter cold-moist stratification period（eight weeks） than other seed lots.展开更多
Eighteen sex hormones in antler velvet were determined by high performance liquid chromatography tandem mass spectrometry.The solid phase extraction was applied to eliminating the matrix effect.The experimental condit...Eighteen sex hormones in antler velvet were determined by high performance liquid chromatography tandem mass spectrometry.The solid phase extraction was applied to eliminating the matrix effect.The experimental conditions were examined and optimized.Under the optimal conditions,the proposed method provides the good linearities and determination limits（0.2―1.0 μg/kg） of the analytes investigated.The recoveries ranging from 72.3% to 149.5% were obtained for the target analytes at two concentration levels.This method was applied to the determination of eighteen sex hormones in different kinds of antler velvet samples and the obtained results are satisfactory.The results indicate that the proposed method is suitable for the determination of sex hormones in antler velvet samples.展开更多
The hydrolysis of velvet bean (Mucuna pruriens) protein in the presence of Alcalase?-Flavourzyme? and Pepsin-Pancreatin was investigated. The results showed that Alcalase?-Flavourzyme? (29.08%) sequential system catal...The hydrolysis of velvet bean (Mucuna pruriens) protein in the presence of Alcalase?-Flavourzyme? and Pepsin-Pancreatin was investigated. The results showed that Alcalase?-Flavourzyme? (29.08%) sequential system catalyzed the hydrolysis most efficiently that Pepsin-Pancreatin (24.78%). In addition, the higher ACE-I inhibitory activity was achieved with the sequential system Alcalase?-Flavourzyme? (33.13%). Furthermore, the concentration of peptides employing an ultrafiltration (UF) system or their purification by gel filtration chromatography showed that the oligomeric peptides with lower molecular weight registered the highest ACE-I inhibitory activity. It has been demonstrated that Mucuna pruriens protein hydrolysates could serve as a source of peptides with ACE inhibitory activity and this activity can be attributed mainly to the mixture of short peptides in the hydrolysate.展开更多
基金This work was supported by the National Natural Science Foundation of China （No. 81302223） and the Medical Scientific Research Foundation of Guangdong Province, China （B2013104）.
文摘Twenty-four-month-old male C57BU6 mice with low serum testosterone levels were used as a late-onset hypogonadism （LOH） animal model for examining the effects of velvet antler polypeptide （VAP） on sexual function and testosterone synthesis. These mice received VAP for 5 consecutive weeks by daily gavage at doses of 100, 200, or 300 mg kg-1 body weight per day （n = 10 mice per dose）. Control animals （n = 10） received the same weight-based volume of vehicle. Sexual behavior and testosterone levels in serum and interstitial tissue of testis were measured after the last administration of VAP. Furthermore, to investigate the mechanisms of how VAP affects sexual behavior and testosterone synthesis in vivo, the expression of steroidogenic acute regulatory protein （STAR）, cytochrome P450 cholesterol side-chain cleavage enzyme （P450scc）, and 3β-hydroxysteroid dehydrogenase （3βHSD） in Leydig cells was also measured by immunofluorescence staining and quantitative real-time PCR. As a result, VAP produced a significant improvement in the sexual function of these aging male mice. Serum testosterone level and intratesticular testosterone （ITT） concentration also increased in the VAP-treated groups. The expression of STAR, P450scc, and 3β-HSD was also found to be enhanced in the VAP-treated groups compared with the control group. Our results suggested that VAP was effective in improving sexual function in aging male mice. The effect of velvet antler on sexual function was due to the increased expression of several rate-limiting enzymes of testosterone synthesis （STAR, P450scc, and 3β-HSD） and the following promotion of testosterone syothesis in vivo.
文摘Background Velvet antler polypeptides (VAPs), which are derived from the antler velvets, have been reported to maintain survival and promote growth and differentiation of neural cells and, especially the development of neural tissues This study was designed to explore the influence of VAPs on neural stem cells in vitro derived from embryonic rat brain Methods Neural stem cells derived from E12 14 rat brain were isolated, cultured, and expanded for 7 days until neural stem cell aggregations and neurospheres were generated The neurospheres were cultured under the condition of different concentration of VAPs followed by immunocytochemistry to detect the differentiation of neural stem cells Results VAPs could remarkablely promote differentiation of neural stem cells and most neural stem cells were induced to differentiate towards the direction of neurons under certain concentration of VAPs Conclusion Neural stem cells can be successfully induced into neurons by VAPs in vitro , which could provide a basis for regeneration of the nervous system
文摘A novel polypeptide having stimulant effect on some cell proliferation was isolated from the velvet antler (Cervus elaphus Linnaeus). The velvet antler polypeptide consists of a single chain of 32 amino acid residues. Amino acid sequence of the polypeptide was identified as: VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM.
文摘天然的丝状真菌是我国重要的微生物资源,能产生大量有益的初生、次生代谢产物,其代谢受到各类调控因子的调控。天鹅绒(Velvet)蛋白家族是真菌中最独特且高度保守的一类特殊调控蛋白,可调控真菌的生长、分化和次生代谢物的生物合成。该文结合国内外的研究进展综述Velvet蛋白家族Velvet A(VeA)、Velvet like B(VelB)在真菌中的结构及其蛋白复合物对生长发育和次生代谢的调控作用,可为进一步探索Velvet蛋白家族调控真菌次生代谢物的生物合成通路、激活沉默基因簇、挖掘新的化合物提供参考。
基金Supported by the Natural Science Foundation of Tianjin (06YFJMJC10500)
文摘Supercritical CO2 (SC-CO2) extraction technology and ultrasonic technology were used to extract two active sex hormones, estradiol and progesterone, and insulin-like growth factor-1 (IGF-1) from antler velvet. The effects of SC-CO2 extraction condition on the extraction yield and content of sex hormones, the ultrasonic extrac-tion condition on the content of IGF-1 and the SC-CO2 extraction condition on the activity remaining of IGF-1 were studied. The optimal conditions were obtained. The experimental results showed that, in presence of 75% ethanol as the co-solvent, the mean yield and content of estradiol and progesterone were 87.67 pg·g-1 and 1224.10 pg·g-1, 12.38 ng·g-1 and 354.06 ng·g-1, respectively, with extraction pressure of 30 MPa, temperature of 35°C, extraction time of 30 min and CO2 consumption of 15 L·g-1 at the flow rate of 2.0 L·min-1. The highest content of IGF-1 was 7425.75 ng·g-1 antler velvet residue, when the pH10 ammonia-ammonium chloride buffer solution was used as the solvent, the ratio of solvent to sample was 20/1 (volume/mass), the extraction temperature was 0-35°C, and the ex-traction time was 4×15 min. Under these conditions, 93.68% activity remaining of IGF-1 in the residue was ob-tained, while little IGF-1 activity exists in traditional residue. The experimental results indicate that the technology of SC-CO2 with co-solvent is of advantage for getting high content sexual hormones and keeping high activity of IGF-1 in the residue, which can not be achieved by traditional extraction methods.
基金Supported by the Natural Science Foundation of Jilin Provincal Science ＆ Technology Department, China（No.201215244） and the Fundamental Research Foundation of Jilin University, China（No.201103232）
文摘Biochemical techniques including ion-exchange chromatography, gel filtration chromatography and re- versed-phase high-performance liquid chromatography（RP-HPLC） were used to isolate and purify the natural poly- peptide from velvet antler（nVAP） of Cervus elaphus（C, elaphus）, which has a molecular weight of 3215.8 and the primary structure of VLSAADKSNVKAAWGKVGGNAPAFGAEALLRM. The homology of the protein sequence in nVAP with known protein sequence is less than 50%, suggesting that nVAP appears to be a new bioactive substance. At a level of 0.4--50 gg/mL, nVAP promotes mitosis in epidermal ceils, chondrocytes and NIH3T3 fibroblasts pri- marily cultured in a significant way. Given that a yield of high-purity nVAP isolated from C. elaphus is 0.001%, nVAP is artificially synthesized to prepare synthetic velvet antler polypeptide（sVAP） according to its primary struc- ture. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） of sVAP shows a single band, and its HPLC spectrum displays a single peak. The matrix-assisted laser desorption ionization time-of-flight mass spectro- metry（MALDI-TOF-MS） was used to identify sVAP to be of a molecular weight of 3200 and the consistency between primary structures of sVAP and nVAE Bioactivity test shows that at a dose of 5--40 μg/mL, sVAP promotes the pro- liferation of primarily cultured epidermal cells and NIH3T3 cell line. From the traditional Chinese medicine theory, velvet antler from Cervus nippon（C, nippon） and velvet antler from C. elaphus are considered as the same medicine, but differences between biochemical base and pharmacological effect of these two velvet antlers have been observed. We compared the total polypeptide mapping of the two velvet antlers, discovering that nVAP is active polypeptide and only exists in the velvet antler of C. elaphus, sVAP is similar to nVAP in physicochemical property and biological activity. These studies extend the possible utility of sVAP to be the promising compound to prepare velvet antler pol
基金the Gorgan University of Agricultural Sciences and Natural Resources for supporting this research
文摘Acer velutinum Boiss is a valuable tree species native to Iran, and its seeds possess physiological dormancy that hampers seedling production in the nursery for large-scale reforestation efforts. The aim of this study was to determine the optimal dormancy breaking treatments for A. velutinum seeds. We conducted a factorial experiment involving six seed lots collected along an elevation gradient from 300 to 1800 m at 300 m interval and four cold-moist stratification periods （0, 4, 8 and 16 weeks） at 4℃ and 70% relative humidity. The result shows that the germination of cold-moist stratified seeds was significantly （p 〈 0.0001） higher than the control for all seed lots. The highest germination capacity was recorded after 16 weeks of cold-moist stratification for all seed lots （68% 88% depending on the seed lot） except those collected from mid altitude sites （600 and 900 m） that germinated equally well （≥ 75%） after 4and 8-week of clod-moist stratification compared to the other seed lots. The mean germination time was significantly shorter （12 to 19 days, depending on the seed lot） for seeds stratified for 16 weeks than for untreated seeds. It can be concluded that： （1） cold-moist stratification for 16 weeks is the best pre-sowing treatment for breaking dormancy in A. velutinum seeds; and （2） seeds should be collected from mid altitude sites （600 and 900 m） to get more than 80% germination within 15 days, and these seed lots even required shorter cold-moist stratification period（eight weeks） than other seed lots.
基金Supported by the Projects of the General Administration of Quality Supervision,Inspection and Quarantine of China(No.2007IK157)
文摘Eighteen sex hormones in antler velvet were determined by high performance liquid chromatography tandem mass spectrometry.The solid phase extraction was applied to eliminating the matrix effect.The experimental conditions were examined and optimized.Under the optimal conditions,the proposed method provides the good linearities and determination limits（0.2―1.0 μg/kg） of the analytes investigated.The recoveries ranging from 72.3% to 149.5% were obtained for the target analytes at two concentration levels.This method was applied to the determination of eighteen sex hormones in different kinds of antler velvet samples and the obtained results are satisfactory.The results indicate that the proposed method is suitable for the determination of sex hormones in antler velvet samples.
文摘The hydrolysis of velvet bean (Mucuna pruriens) protein in the presence of Alcalase?-Flavourzyme? and Pepsin-Pancreatin was investigated. The results showed that Alcalase?-Flavourzyme? (29.08%) sequential system catalyzed the hydrolysis most efficiently that Pepsin-Pancreatin (24.78%). In addition, the higher ACE-I inhibitory activity was achieved with the sequential system Alcalase?-Flavourzyme? (33.13%). Furthermore, the concentration of peptides employing an ultrafiltration (UF) system or their purification by gel filtration chromatography showed that the oligomeric peptides with lower molecular weight registered the highest ACE-I inhibitory activity. It has been demonstrated that Mucuna pruriens protein hydrolysates could serve as a source of peptides with ACE inhibitory activity and this activity can be attributed mainly to the mixture of short peptides in the hydrolysate.
