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1.
Article in English | WPRIM | ID: wpr-741644

ABSTRACT

Eleven steroid hormones (SHs: androstene-3,17-dione, estrone, β-estradiol, α-estradiol, testosterone, dehydroepiandrosterone, 17á-hydroxyprogesterone, medroxyprogesterone, megestrol acetate, progesterone, and androsterone) were detected from New Zealand deer (Cervus elaphus var. scoticus) velvet antler (NZA, 鹿茸). A method for the quantification of eleven SHs was established by using ultraperformance liquid chromatography (UPLC)-MS/MS. The linearities (R² > 0.991), limits of quantification (LOQ values, 0.3 ng/mL to 23.1 ng/mL), intraday and interday precisions (relative standard deviation: RSD 0.999), LOQ values (30 ng/mL to 350 ng/mL), intraday and interday precisions (RSD < 1.93%), and recovery rates (97.2% to 103.5%) for the three 7-O-CSs were determined. These quantitative methods are accurate, precise, and reproducible. As a result, it is suggested that the five steroid compounds of androstene-3,17-dione, androsterone, 7-ketocholesterol, 7α-hydroxycholesterol, and 7β-hydroxycholesterol could be marker steroids of NZA. These methods can be applied to quantify or standardize the marker steroids present in NZA.


Subject(s)
Androsterone , Animals , Antlers , Chromatography, Liquid , Deer , Dehydroepiandrosterone , Estrone , Medroxyprogesterone , Megestrol Acetate , Methods , New Zealand , Progesterone , Steroids , Testosterone
2.
Article in English | WPRIM | ID: wpr-648769

ABSTRACT

Podostroma cornu-damae is a rare, deadly fungus. However, it can be easily mistaken for antler Ganoderma lucidum. In this case report, two patients made tea with the fungus and drank it over a 2-week period. Both patients presented with bicytopenia, and one patient had desquamation of the palms and soles. Both were treated with prophylactic antibiotics and granulocyte colony-stimulating factor. One patient was admitted to the intensive care unit and received a platelet transfusion. Both patients were discharged without complications. Podostroma cornu-damae infections caused by intoxication were successfully treated using our treatment strategy, which consisted of prophylactic antibiotics, platelet transfusion, and granulocyte colony-stimulating factor. We believe this report can guide future treatment.


Subject(s)
Agaricales , Alopecia , Animals , Anti-Bacterial Agents , Antlers , Fungi , Granulocyte Colony-Stimulating Factor , Humans , Intensive Care Units , Pancytopenia , Platelet Transfusion , Poisoning , Reishi , Tea
3.
Article in Korean | WPRIM | ID: wpr-13822

ABSTRACT

The effects of antlers have long been known in traditional Asian medicine. However, few studies have investigated the effects of antlers on immunity. In this study, we investigated whether fermented antler extract (FAE) has immunomodulatory effects on spleen cells. FAE enhanced the activity of spleen cells in a concentration dependent manner compared to antler extract. Interestingly, FAE significantly increased the production of interleukin-12, a representative cytokine of cell-mediated immunity, while it marginally increased that of tumor necrosis factor-alpha. Flow cytometry analysis demonstrated that FAE can protect spleen cells from spontaneous cell death without a significant proportional change in subsets, mainly lymphocytes. Taken together, the results of the present study showed that FAE has beneficial effects on spleen cells, a major type of immune cell, indicating that it can function as an immunomodulator without significant cytotoxicity. These data may broaden the use of FAE in basic research and clinical areas.


Subject(s)
Animals , Antlers , Asians , Cell Death , Flow Cytometry , Humans , Immunity, Cellular , Immunomodulation , Interleukin-12 , Lymphocytes , Spleen , Tumor Necrosis Factor-alpha
4.
Acta Pharmaceutica Sinica ; (12): 1356-1361, 2015.
Article in Chinese | WPRIM | ID: wpr-320077

ABSTRACT

In order to authenticate the components of antler powder in the market, DNA barcoding technology coupled with cloning method were used. Cytochrome c oxidase subunit I (COI) sequences were obtained according to the DNA barcoding standard operation procedure (SOP). For antler powder with possible mixed components, the cloning method was used to get each COI sequence. 65 COI sequences were successfully obtained from commercial antler powders via sequencing PCR products. The results indicates that only 38% of these samples were derived from Cervus nippon Temminck or Cervus elaphus Linnaeus which is recorded in the 2010 edition of "Chinese Pharmacopoeia", while 62% of them were derived from other species. Rangifer tarandus Linnaeus was the most frequent species among the adulterants. Further analysis showed that some samples collected from different regions, companies and prices, contained adulterants. Analysis of 36 COI sequences obtained by the cloning method showed that C. elaphus and C. nippon were main components. In addition, some samples were marked clearly as antler powder on the label, however, C. elaphus or R. tarandus were their main components. In summary, DNA barcoding can accurately and efficiently distinguish the exact content in the commercial antler powder, which provides a new technique to ensure clinical safety and improve quality control of Chinese traditional medicine


Subject(s)
Animals , Antlers , DNA Barcoding, Taxonomic , Deer , Medicine, Chinese Traditional , Polymerase Chain Reaction , Powders , Quality Control
5.
Acta Pharmaceutica Sinica ; (12): 1637-1644, 2015.
Article in Chinese | WPRIM | ID: wpr-320030

ABSTRACT

This study was designed to use iTRAQ technology coupled with 2D LC-MS/MS to study the comparative proteomics of different processing technology for pilose antler. 1015 proteins were identified with 2D LC combined with MOLDI TOF/TOF mass spectrometry. Comparative analysis with Protein Pilot (Version 4.5) revealed that 87 proteins were changed (P ≤ 0.05, the ratio of > 1.50 or < 0.60 as the threshold selection of difference proteins), of which 24 were up regulated and 33 were down regulated in the traditional frying process (TFP) compared with the fresh pilose antler (P ≤ 0.05). 7 significant different proteins (P ≤ 0.001), most of these significantly changed proteins were found to be involved in calcium ion binding and ATP binding associated with human healthy. Freeze drying with protective agent (FDP) (Trehalose) can improve the content of significantly different proteins (P ≤ 0.001) including Collagen alpha-1 (XII) chain (COL12A1) and Collagen alpha-1 (II) chain (COL2A1). The significant function involves in platelets activating, maintenance of spermatogonium, and disorder expression in tumor cells. The functional annotation by Hierarchical clustering and GO (gene ontology) showed that the main molecule functions of the proteins significantly changed in these processes were involved in binding (52.7%), catalytic (25.3%), structural molecule and transporter (6.6%).


Subject(s)
Animals , Antlers , Chemistry , Chromatography, Liquid , Collagen , Chemistry , Down-Regulation , Freeze Drying , Gene Expression Regulation , Proteomics , Tandem Mass Spectrometry , Technology, Pharmaceutical , Methods , Up-Regulation
6.
Article in Chinese | WPRIM | ID: wpr-330189

ABSTRACT

High resolution melting (HRM) , an important technology for genotyping and mutation scanning, has broad prospects in the authenticity of traditional Chinese medicine. This paper selected universal CO I primers and used HRM to establish a new method for authenticity of Hairy Antler. PCR was conducted at the annealing temperature of 60 °C and 45 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further optimized. The results showed that the Tm values of Cervus nippon were (81.96 ± 0.07), (84.51 ± 0.03) °C and Cervus elaphus was(82.58 ± 0.13), (85.95 ± 0.05) °C with 10-100 mg · L(-1) DNA template, 0.2 µLmol · L(-1) primer, 2.0 mmol · L(-1) Mg2+. This method can authenticate of hairy antler and is simple, fast, high-throughput, visualization.


Subject(s)
Animals , Antlers , Chemistry , DNA , Chemistry , Genetics , DNA Primers , Genetics , Deer , Classification , Genetics , Genotype , Medicine, Chinese Traditional , Reference Standards , Polymerase Chain Reaction , Transition Temperature
7.
Article in English | WPRIM | ID: wpr-34496

ABSTRACT

BACKGROUND/OBJECTIVES: We examined the chemical composition and the effect of fermented deer antler on hematopoietic factors in bone marrow cells. MATERIALS/METHODS: For the preparation of fermented deer antler extract (FAB), fermentation was carried out using Bacillus subtilis at 30degrees C for 7 days. The hematopoietic effect of FAB was investigated hematopoietic factors in marrow cells. RESULTS: The contents of total sugar, sulfated glycosaminoglycans, and uronic acid and the dry weight gradually increased with fermentation time. The sialic acid content (from 0.14 mg/mL to 0.54 mg/mL) was the highest on the 4th day of fermentation after which it decreased. The proliferating activity of bone marrow cells increased with fermentation times. The levels of various hematopoietic growth factors were determined to verify the beneficial effect of deer antler extract fermented by B. subtilis on hematopoiesis. FAB increased the number of stem cell factors and granulocyte colony-stimulating factor in bone marrow cells. In addition, FAB augmented the burst-forming unit erythroid and total colonies in splenocyte-conditioned medium compared with non-fermented antler extract (NFA). However, FAB did not affect the mRNA levels of erythropoietin, an important factor for erythropoiesis. CONCLUSIONS: FAB, like NFA, did not directly affect hematopoiesis, but contributed to hematopoiesis by stimulating the production of hematopoietic factors.


Subject(s)
Animals , Antlers , Bacillus subtilis , Bacillus , Bone Marrow Cells , Bone Marrow , Deer , Erythropoiesis , Erythropoietin , Fermentation , Glycosaminoglycans , Granulocyte Colony-Stimulating Factor , Hematopoiesis , Intercellular Signaling Peptides and Proteins , N-Acetylneuraminic Acid , RNA, Messenger , Stem Cell Factor
8.
Int. j. morphol ; 32(1): 12-15, Mar. 2014. ilus
Article in English | LILACS | ID: lil-708715

ABSTRACT

The objective of this study was to investigate asymmetry in antlers of Caspian Red Deer or maral (Cervus elaphus sibiricus) from two different genetic origins and maintained under similar conditions. Eighteen male Caspian Red Deer aged 24 months were studied. Nine animals belonged to the local Kazakh population and nine were directly imported Russian deer. The following data were obtained for right and left antlers: wet weight, stem length and circle and 1st, 2nd and 3rd shoot lengths. To obtain the values of antler asymmetries we used the absolute differences between the value of each trait on left and right sides. According to values of asymmetries obtained, the two populations neither showed differences from each other nor exhibited consistent directional trends in mean measures. Therefore, it could be concluded that Russian marals have adapted well to environmental conditions, presenting no different levels of environmental stress in relation to local Kazakh ones.


El objetivo de este estudio fue investigar la asimetría en las astas del Ciervo Rojo del Mar Caspio o Maral (Cervus elaphus sibiricus) en dos rebaños de origen genético diferente y mantenidos bajo condiciones similares. Se estudiaron 18 ciervos rojos del mar Caspio (machos) de 24 meses de edad. Nueve de los 18 animales pertenecían a la población nativa de Kazajistán y nueve fueron importados directamente desde Rusia. Se obtuvieron los siguientes datos para las astas derecha e izquierda: peso húmedo, longitud del tallo, circunferencia y primera, segunda y tercera longitud de los cuernos. Para obtener los valores de asimetrías de las astas se registraron las diferencias absolutas entre el valor de cada rasgo, izquierdo y derecho. De acuerdo a los valores obtenidos de las asimetrías, no se registraron diferencias entre ambas poblaciones, tampoco se observaron tendencias direccionales consistentes en el promedio de medidas. El ciervo rojo ruso se ha adaptado bien a las condiciones ambientales y no presentó niveles de estrés ambiental diferentes en relación con los animales nativos de Kazajistán.


Subject(s)
Male , Antlers/anatomy & histology , Deer/anatomy & histology
9.
Article in Chinese | WPRIM | ID: wpr-312843

ABSTRACT

<p><b>OBJECTIVE</b>To observe the effect of single herb pilose antler (PA) on the expression of Smad2 and Smad3 in the cartilage of osteoarthritis (OA) rats.</p><p><b>METHODS</b>One hundred 3-month old female healthy SD rats, (200 +/- 20) g, were recruited and routinely fed for 1 week. They were randomly divided into 5 groups, i.e., the low dose PA group, the high dose PA group, the normal saline control group, the model group, and the normal control group, 20 in each group. The model was prepared using classic Hulth method except the normal control group. After 6-week modeling, the model was confirmed successful by pathologic observation. PA at 0.021 g/100 g and 0.084 g/1 00 g was given by gastrogavage to rats in the low dose PA group and the high dose PA group respectively. Normal saline was administered to those in the normal saline control group. No treatment was given to rats in the normal control group and the model group. Bilateral knee cartilages were harvested at week 2,4, and 6. mRNA and protein expressions of Smad2 and Smad3 were detected by immunohistochemical assay, fluorescent quantitative PCR, and Western blot.</p><p><b>RESULTS</b>OA model was successfully prepared by pathological observation. Results of immunohistochemical assay showed that Smad2 and Smad3 expressed extensively in the cartilage, and located inside the chondrocyte membrane. Compared with the model group, mRNA expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 2, 4, and 6, showing statistical difference (P < 0.05). Compared with the same group at week 4 after gastrogavage, mRNA expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.05). Compared with the model group, protein expression of Smad2 and Smad3 obviously increased in the chondrocytes of the low dose PA group and the high dose PA group at week 2 and 4, showing statistical difference (P < 0.01). Compared with the same group at week 2 after gastrogavage, protein expression of Smad2 and Smad3 obviously increased in the low dose PA group and the high dose PA group at week 4, showing statistical difference (P < 0.01). Compared with the same group at week 4 after gastrogavage, protein expression of Smad2 and Smad3 obviously decreased in the low dose PA group and the high dose PA group at week 6, showing statistical difference (P < 0.01).</p><p><b>CONCLUSIONS</b>(1) The pilose antler could repair cartilages by regulating mRNA and protein expressions of Smad2 and Smad3. (2) Up-regulating mRNA and protein expressions of Smad2 and Smad3 might be one of important mechanisms for the pathogenesis of OA.</p>


Subject(s)
Animals , Antlers , Chemistry , Cartilage , Cell Biology , Metabolism , Chondrocytes , Metabolism , Female , Medicine, Chinese Traditional , Osteoarthritis , Drug Therapy , Metabolism , Rats , Rats, Sprague-Dawley , Smad2 Protein , Metabolism , Smad3 Protein , Metabolism
10.
Article in English | WPRIM | ID: wpr-310893

ABSTRACT

Deer velvet antler is the only mammal organ which can continuous regenerate. Currently, international scholars are interested in antler that is defined as a perfect regeneration model of neuro, blood vessel, connective tissue, cartilage, and bones. In 1986, we started to study the separation of active protein and peptide of fresh velvet antler using classic biochemical methods. After entering the 21st century, we further investigated the differentiation of antler proteome from different growth periods using advance differential proteomics approach, and unveiled the correlation between the proteome difference and life cycle. The international antler research has entered the stage of molecular biology, and will no doubt have a profound impact on the modern biomedical fields, such as regenerative medicine, organ degeneration and dysplasia, trauma medicine and anti-inflammatory treatment, growth factor research, as well as creation of new medical thinking.


Subject(s)
Animals , Antlers , Chemistry , Deer , Humans , Medicine, Chinese Traditional , Peptides , Pharmacology , Regenerative Medicine
11.
Article in English | WPRIM | ID: wpr-727669

ABSTRACT

The purpose of this study is to characterize the effects of KHG26792 (3-(naphthalen-2-yl(propoxy) methyl)azetidine hydrochloride), a potential skin whitening agent, on melanin synthesis and identify the underlying mechanism of action. Our data showed that KHG26792 significantly reduced melanin synthesis in a dose-dependent manner. Additionally, KHG26792 downregulated microphthalmia-associated transcription factor (MITF) and tyrosinase, the rate-limiting enzyme in melanogenesis, although tyrosinase was not inhibited directly. KHG26792 activated extracellular signal-regulated kinase (ERK), whereas an ERK pathway inhibitor, PD98059, rescued KHG26792-induced hypopigmentation. These results suggest that KHG26792 decreases melanin production via ERK activation. Moreover, the hypopigmentary effects of KHG26792 were confirmed in a pigmented skin equivalent model using Cervi cornus Colla (deer antler glue), in which the color of the pigmented artificial skin became lighter after treatment with KHG26792. In summary, our findings suggest that KHG26792 is a novel skin whitening agent.


Subject(s)
Animals , Antlers , Cornus , Hypopigmentation , MAP Kinase Signaling System , Melanins , Microphthalmia-Associated Transcription Factor , Monophenol Monooxygenase , Phosphotransferases , Skin Lightening Preparations , Skin , Skin, Artificial
12.
Article in Chinese | WPRIM | ID: wpr-346471

ABSTRACT

<p><b>OBJECTIVE</b>To determine the contents of amino acids in Cornu Cervi Pantotrichum of different specifications for controlling the quality.</p><p><b>METHOD</b>The contents of 18 kinds of amino acids were determined by amino acid analyzer.</p><p><b>RESULT</b>The correlation coefficients of 18 kinds of amino acids were all greater than 0.997, the average recovery were all between 99.1%-108.1% with RSDs less than 2.0%. All Cornu Cervi Pantotrichum samples of 29 different specifications contained 17 kinds of amino acids and 7 kinds of essential amino acids. The content of total amino acids in wax slices is relative higher. The content in first born antlers is higher than that in reborn antlers.</p><p><b>CONCLUSION</b>This method is suitable for the determination of amino acids in Cornu Cervi Pantotrichum, it provides good reference for the quality control of Cornu Cervi Pantotrichum.</p>


Subject(s)
Amino Acids , Animals , Antlers , Chemistry , Deer , Medicine, Chinese Traditional
13.
Article in Chinese | WPRIM | ID: wpr-321860

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of Pilose antler polypeptide on apoptosis of chondrocyte and related cytokines in experimental knee osteoarthritis.</p><p><b>METHODS</b>Totally 64 New Zealand White rabbits of 6 months old were randomly divided into 2 groups:normal group(n=8)and model group (n=56). Model group was surgically induced into knee osteoarthritis model by method of Hulth. After successful modeling,the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group (n=24) and control group (n=24). Pilose antler polypeptide-treatment group received 0.5 ml intraarticular injection of Pilose antler polypeptide dilution liquid once per 2 days for 30 days while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples and synovial fluid were collected respectively. The morphological changes of articular cartilage under optical microscope and the structural change of chondrocyte were observed by transmission electron microscopy. The levels of interleukin-1beta and tumor necrosis factor-alpha in synovial fluid was detected by Enzyme-linked Immunosorbent Assay.</p><p><b>RESULTS</b>Along with the extending of time, articular cartilage degenerated gradually and chondrocytes apoptosis increased significantly. On days 7,15 and 30 after intervention, the chondrocyte apoptosis index of the Pilose antler polypeptide-treatment group were (20.30 +/- 1.23), (28.60 +/- 2.37), (37.10 +/- 1.82) and those of control group were (31.50 +/- 2.44), (34.40 +/- 1.77), (42.30 +/- 2.33). There were significant differences between them (P<0.05). At the same time, the chondrocyte apoptosis index of the Pilose antler polypeptide-treatment group were lower than those of control group,which had a statistical significance (P<0.05). On days 7,15 and 30 after intervention, the levels of interleukin-1beta in synovia fluid of Pilose antler polypeptide-treatment group were (15.81 +/- 1.26), (12.59 +/- 1.42), (9.57 +/- 0.92) microg/L and the level of tumor necrosis factor-alpha were (48.47 +/- 2.64), (43.46 +/- 1.33), (40.96 +/- 1.05) microg/L, with statistical differences(P<0.05). The levels of interleukin-1beta in synovia fluid of control group were (18.92 +/- 1.83), (20.25 +/- 2.76), (22.13 +/- 2.24) microg/L and the levels of tumor necrosis factor-alpha were (57.92 +/- 2.12), (60.25 +/- 1.48), (63.35 +/- 2.15) microg/L. At the same period,the levels of interleukin-1beta and tumor necrosis factor-alpha were lower than those of the control group,which had a statistical significance (P<0.05).</p><p><b>CONCLUSION</b>Pilose antler polypeptide can inhibit chondrocytes apoptosis, decrease the levels of interleukin-1beta and tumor necrosis factor-alpha and delay the degeneration of articular cartilage to some extent.</p>


Subject(s)
Animals , Antlers , Chemistry , Apoptosis , Chondrocytes , Pathology , Female , Interleukin-1beta , Male , Osteoarthritis, Knee , Drug Therapy , Allergy and Immunology , Pathology , Peptides , Pharmacology , Rabbits , Tumor Necrosis Factor-alpha
14.
Article in Chinese | WPRIM | ID: wpr-248881

ABSTRACT

<p><b>OBJECTIVE</b>To observe the influence of Pilose antler polypeptide on the glycosaminoglycan and type II collagen in the articular cartilage in experimental knee osteoarthritis.</p><p><b>METHODS</b>Totally 64 New Zealand white rabbits of 6 months old were randomly divided into 2 groups:normal group (n = 8) and model group (n = 56). Model group was surgically induced into osteoarthritis model by method of Hulth. After successful modeling, the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group and control group, 24 rabbits in each group. Pilose antler polypeptide-treatment group received 0.5 ml intra-articular injection of Pilose antler polypeptide dilution liquid once in per 2 days for 30 days, while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples were collected respectively. The content of glycosaminoglycan in articular cartilage was observed by toluidine blue staining and the expression of type II collagen in cartilage matrix was detected by immunohistochemical staining.</p><p><b>RESULTS</b>Along with the prolonging of time, the content of glycosaminoglycan and type II collagen in cartilage matrix of the Pilose antler polypeptide-treatment group and control group decreased gradually. On days 7, 15 and 30 after intervention, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group were (312.06 +/- 14.12), (273.31 +/- 12.42) and (248.34 +/- 10.41), which had statistically significant differences. Integrated optical density of the type II collagen positive area in cartilage matrix of the control group were (253.47 +/- 15.53), (215.67 +/- 9.72) and (160.01 +/- 13.23), which had statistically significant differences. At the same period, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group was higher than that of control group, which had statistically significant difference.</p><p><b>CONCLUSION</b>Pilose antler polypeptide can inhibit reduction of the glycosaminoglycan and type II collagen in cartilage matrix and delay the degeneration of articular cartilage.</p>


Subject(s)
Animals , Antlers , Chemistry , Metabolism , Collagen Type II , Metabolism , Disease Models, Animal , Female , Glycosaminoglycans , Metabolism , Humans , Male , Osteoarthritis, Knee , Drug Therapy , Metabolism , Peptides , Metabolism , Pharmacology , Rabbits
15.
Article in Chinese | WPRIM | ID: wpr-344637

ABSTRACT

<p><b>OBJECTIVE</b>To study reverse effect of the oxidative damage on cartilage cells of velvet antler polypeptides (VAPS), and to investigate the main mechanism of VAPS to protect cartilage cells through antioxidant.</p><p><b>METHODS</b>Fifteen Japanese white rabbits of 5-month-old were selected in this study. Animal model was established by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut, articular cartilage cell cultured in vitro. Cells in the sham operation group was the normal control group, osteoarthritis cartilage cells in the model groups were added VAPS 6.25, 12.5, 25 microg/ml respectively. A group of animals were sacrificed every week form the ninth weeks(two months) and the cartilage cells were isolated and cultured. For 8 weeks,the reactive oxygen species level in chondrocytes were detected by DCFH-DA, the content of NO, SOD and GSH-Px in cell culture supernatant were detected by Griess method.</p><p><b>RESULTS</b>DCFH-DA detection of intracellular reactive oxygen species was (5.46 +/- 0.46)in the control group, (12.08 +/- 0.74) in the model groups. The model group compared with the control group by t test with the P value less than < 0.001. DCFH-DA detection of intracellular reactive oxygen species was (9.81 +/- 0.59)in VAPS 6.25 microg/ ml group, (7.83 +/- 0.63) in the VAPS 12.5 microg/ml group, (6.89 +/- 0.71) in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.05). The content of NaNO2, SOD and GSH-Px in osteoarthritis model group was (5.60 +/- 0.45) microM, (38.56 +/- 12.53) U/ml and (151.90 +/- 25.60) U, as compared with control group there were statistically significant difference (P < 0.001, P < 0.05); The content of NaNO2 was (4.34 +/- 0.39), M in VAPS 6.25 microg/ml group, (3.67 +/- 0.36) microM in the VAPS 12.5 microg/ml group, (3.20 +/- 0.27) microM in the VAPS 25 microg/ml group, as compared with model group there were statistically significant difference (P < 0.01). The content of SOD was (49.91 +/- 5.77) U/ml in VAPS 6.25 microg/ml group, (54.05 +/- 5.27) U/ml in the VAPS 12.5 microg/ml group, (57.44 +/- 5.70) U/ml in the VAPS 25 microg/mL group, as compared with model group there was statistically significant (P < 0.05). The content of GSH-Px was (172.50 +/- 18.65) U in VAPS 6.25 microg/ml group, (202.10 +/- 21.60) U in the VAPS 12.5 microg/ml group, (315.80 +/- 10.50) U in the VAPS 25 microg/ml group, the VAPS 12.5 microg/mL group and VAPS 25 microg/ml group was compared with model group, there were statistically significant difference (P < 0.01).</p><p><b>CONCLUSION</b>The VAPS have antioxidative damage effect of osteoarthritis cartilage cells within a certain range and dose-dependent manner. It may be the main mechanism for velvet antler polypeptides to treat osteoarthritis.</p>


Subject(s)
Animals , Antlers , Chemistry , Cartilage , Metabolism , Pathology , Female , Glutathione , Blood , Male , Nitric Oxide , Blood , Osteoarthritis , Blood , Metabolism , Pathology , Oxidative Stress , Peptides , Pharmacology , Rabbits , Reactive Oxygen Species , Metabolism , Superoxide Dismutase , Blood
16.
Acta Pharmaceutica Sinica ; (12): 1526-1529, 2011.
Article in Chinese | WPRIM | ID: wpr-323090

ABSTRACT

An in vitro detection method of the gastrointestinal absorption of Pilose Antler protein was established for mixed protein activity. Five bands of protein with molecular weight of 17.8-160 kD derived from the Pilose Antler were extracted and sufficiently labeled with FITC (FITC-PE). The stability and variation of FITC-PE in gastrointestinal circumstances were detected by native polyacrylamide gel electrophoresis and confocal laser scanning microscope. Results showed that the main component of FITC-PE kept invariant after being reacted with artificial gastric fluid and artificial intestinal fluid. The fluorescence signal was detected 20 min after administration in the valgus intestinal purse experiment, and three kinds of protein, with molecular weight of 45, 25, and 17.8 kD, were detected in the mixture of absorbent protein. The research laid the foundation for the further in vivo study of Pilose Antler protein. Meanwhile, it would be an in vitro screening method for the absorption, distribution and metabolism of mixed protein from traditional Chinese medicine.


Subject(s)
Animals , Antlers , Chemistry , Deer , Fluorescein-5-isothiocyanate , Intestinal Absorption , Intestinal Mucosa , Metabolism , Male , Materia Medica , Chemistry , Pharmacokinetics , Microscopy, Confocal , Molecular Weight , Native Polyacrylamide Gel Electrophoresis , Proteins , Chemistry , Pharmacokinetics , Rats , Rats, Wistar , Stomach , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-281721

ABSTRACT

<p><b>OBJECTIVE</b>To research the effect of anthraquinone extracted from rubiqmnone on growth inhibition, introduction apoptosis and expression of relative gene of bcl-2 of hepatocarcinoma cell SMMC 7721, and provide the effective target of gene therapy.</p><p><b>METHOD</b>The growth inhibition effect was detected by MTF. Morpholgy, DNA agarose gel electrophoresis and flow cytometry were used to observe the cell apoptosis. The effect of anthraquinone on bcl-2mRNA expression was analyzed by RT-PCR.</p><p><b>RESULT</b>Anthraquinone could inhibit the growth of hepatocarcinoma cell SMMC 7721. The typical apoptosis cells were found by inverted microscope and common microscope. DNA agarose gel electrophoresis showed a typical apoptosis strip. G1 period of cell cycle had apoptosis peak of abnormal diploid by PI simple stain, and cell cycle stopped at G1 period. The apoptosis fuction of anthraquinone introdution hepatocarcinoma cell was further certified by Annexin V-FITC/PI. Anthraquinone could decrease the expression of bcl-2mRNA by RT-PCR.</p><p><b>CONCLUSION</b>Anthraquinone can significantly inhibit growth of hepatocarcinoma cell and induce apoptosis. The mocular mechanism may be related to down-regulating the expression of bcl-2 gene.</p>


Subject(s)
Animals , Anthraquinones , Therapeutic Uses , Antlers , Chemistry , Apoptosis , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Cell Proliferation , Deer , Gene Expression , Genes, bcl-2 , Physiology , Humans , Liver Neoplasms , Pathology , Reverse Transcriptase Polymerase Chain Reaction
18.
Article in Chinese | WPRIM | ID: wpr-279385

ABSTRACT

Lyophylized antler powder was hydrolyzed by pepsin and trypsin separately and also simultaneously to give hydrolysates with special physical activities. Complete hydrolysis peptides with MW lower than 1 x 10(3) were collected for assay of angiotensin I-converting enzyme (ACE) inhibitory activity, antioxidant activity and proliferative activity toward UMR-106 osteoblast cells. The results of the experiments revealed that all hydrolysates exhibited potent hydroxyl radical scavenging activity with an IC50 value less than 1 mg/ml which was much lower than the value of 5.5 g x L(-1) for vitamin C. The peptic and peptic tryptic hydrolysates demonstrated strong angiotensin I-converting enzyme (ACE) inhibitory activity. The tryptic hydrolysate increased the proliferation of the UMR-106 cells by 73.43%. The results verified the traditional use of antler in bone-strengthening, anti-aging. The exploratory studies on the ACE inhibitory activity of antler hydrolysates indicated that the hydrolysates might be potentially useful in prevention and treatment of hypertension. Further purification of peptides contributing to the antioxidant activity, angiotensin I-converting enzyme-inhibitory activity and proliferative activity toward osteoblasts from antler hydrolysates is warranted.


Subject(s)
Angiotensin-Converting Enzyme Inhibitors , Metabolism , Animals , Antioxidants , Metabolism , Antlers , Chemistry , Metabolism , Biphenyl Compounds , Metabolism , Cell Proliferation , Deer , Endopeptidases , Metabolism , Free Radical Scavengers , Hydrolysis , Hypertension , Pepsin A , Metabolism , Peptides , Pharmacology , Peptidyl-Dipeptidase A , Metabolism , Picrates , Metabolism , Trypsin , Metabolism
19.
Annals of Dermatology ; : 173-179, 2010.
Article in English | WPRIM | ID: wpr-54706

ABSTRACT

BACKGROUND: Fibroblasts produce many components of the extracellular matrix (ECM) and so they contribute to the maintenance of connective tissue integrity. OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: These results suggest that VAE has a potential to stimulate ECM production, and VAE may be applicable for maintaining the skin's texture.


Subject(s)
Animals , Antlers , Blotting, Western , Cell Movement , Collagen Type I , Connective Tissue , Elastin , Enzyme-Linked Immunosorbent Assay , Extracellular Matrix , Fibroblasts , Fibronectins , Gene Expression , Humans , Oligonucleotide Array Sequence Analysis , Phosphorylation , Protein Kinases
20.
Article in Chinese | WPRIM | ID: wpr-346982

ABSTRACT

<p><b>OBJECTIVE</b>To establish a convenient, quick and accurate molecular method for the identification of crude drugs of antlers due to the difficult discrimination between the genuine antler and its adulterants.</p><p><b>METHOD</b>According to the alignment analysis of full length sequences of Cyth gene from closely relate species of Cervus, one pair of allele-specific diagnostic PCR primers was designed. Factors such as annealing temperature, dosage of polymerase, times of cycles and dosage of template DNA that influence the PCR results were also investigated.</p><p><b>RESULT</b>Based on the study mentioned above, about 323 bp positive band was amplified under the annealing temperature of 65 degrees C in the total volume of 25 microL PCR reaction using the genuine antler DNA as the template. Sequencing results proved that the positive band was the fragment of Cytb gene from both C. elaphus Linnaeus and C. nippon Temminck.</p><p><b>CONCLUSION</b>The established method, with higher specificity and reproducibility, could accurately differentiate genuine antler from its adulterants and would be widely used in Cervus related Chinese crude drugs' identification.</p>


Subject(s)
Alleles , Animals , Antlers , Chemistry , China , DNA Primers , Genetics , Deer , Genetics , Molecular Sequence Data , Polymerase Chain Reaction , Methods , Species Specificity
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