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Objective: To investigate the chemical constituents from the stems and leaves of Dioscorea opposita. Methods: The compounds were isolated and purified by various column chromatographies, and their structures were identified by physiochemical properties and spectroscopic data. The effects of the compounds on the proliferation of human breast cancer MCF-7 cells and human liver cancer HepG2 cells were investigated. Results: Twenty compounds were isolated from the 50% acetone extract of the stems and leaves of D. opposita, which were identified as 1H-indazole (1), 1H-indole-3-carboxaldehyde (2), 1H-indole-3- carboxylic acid (3), 3-(2-oxopropyl)-3-hydroxy-indolin-2-one (4), thymidine (5), uridine (6), 3-hydroxy-3-(2-oxopropyl)-2,4-(1H,3H)- quinolinedione (7), hematinic acid (8), allantoin (9), 2-ethyl-3-methyl-maleimide-N-β-D-glucopyranoside (10), paulownin (11), (+)-8-hydroxypinoresinol (12), (+)-syringaresinol (13), (-)-dihydrodehydrodiconiferyl alcohol (14), (7R,8S)-dihydrodehydro- diconiferyl alcohol-4-O-β-D-glucopyranoside (15), (2E,6S)-6,7-dihydroxy-3,7-dimethyl-2-octenoic acid (16), (2E,4S)-4-hydroxy-2- nonenoic acid (17), (2E,6S)-6-hydroxy-2,6-dimethyl-2,7-dienoic acid (18), amarantholidoside IV (19), (9Z,11E)-13-methoxy- 9,11-octadecadienoic acid methyl ester (20), and their effects on proliferation of MCF-7 cells and HepG2 cells were investigated. compounds 1, 4, 8, 11-15, 18, 20 at a dose of 25μmol/L inhibited the proliferation of MCF-7 cells, and compounds 1, 4, 8, 12-14, 18, 20 at a dose of 25 μmol/L inhibited the proliferation of HepG2 cells. Conclusion: Compounds 1, 3-5, 10-12 and 15-20 are isolated from this plant for the first time, compounds 1, 4,8, 11-15, 18, 20 inhibited the the proliferation of MCF-7 and HepG2 cellssignificantly at certain concentration, showing protent antitumor activity.
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OBJECTIVE: To compare the differences of macrophages phagocytic activities between Dioscorea opposita Thunb. cv. Tiegun(abbreviated as TG) and Dioscorea opposita Thunb. but not Tiegun(abbreviated as NTG). METHODS: CCK-8 assay was used to test the cytotoxicity. On the basis of non-toxic dose, the high content screening(HCS) cell imaging analysis was applied to test the phagocytic ability of RAW264.7 cells engulfing GFP-E. coli in vitro, and the phagocytic rates between different kinds of samples at various concentrations were calculated. Furthermore, the biological potency was measured according to the qualitative response parallel method to better evaluate the difference of TG and NTG, by translating phagocytic rates into biological potency. RESULTS: At the range of 0.695 - 5.56 mg(crude drug) •mL-1, all 11 batches samples have no toxicity. The results of HCS displayed that every sample could promote cell phagocytic activity in varying degrees at a certain concentration. RAW264.7 cells could engulf the GFP-E. coli, and the images of HCS reflected the situation of phagocytosis clearly. In addition, the results of biopotency showed that the biopotency value of different samples was between 39.56 to 100, and the average biopotency value of TG was significantly higher than NTG (P < 0.05). CONCLUSION: In vitro experiments showed that all the samples could promote the phagocytic activity of RAW264.7 cells at different degrees. And the average biopotency value of TG was significantly higher than NTG, which is consistent with the saying that "TG has a better quality".
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Objective: To establish a high content screening (HCS) method by testing the phagocytic function of RAW 264.7, and to evaluate the effect of Dioscorea opposita on cell proliferation and phagocytosis in vitro. Methods: CCK-8 was used to measure the proliferation of RAW 264.7, and HCS was helpful to determine the ability of RAW 264.7 to engulf GFP-E.coli. Results: The best method was established by examining the time of administration, the amount of bacteria added, and the time of bacterial stimulation: administration time of 12 h, 50 times of bacteria, and stimulation time of 1.5 h. Then, the repeatability of this method was tested, and the RSD value was 2.31%. Compared with the group without drugs, at the concentration of 0.156-1.25 mg/mL, the water extract of D. opposita could promote the proliferation, and improve the average phagocytosis rate of every single RAW264.7 cell. And the samples had the strongest effect on proliferation and phagocytosis when the concentration was 1.25 mg/mL. Conclusion: An HCS method is established and firstly used in D. opposita. HCS has the advantage of accurate, intuitive, and high throughput. D. opposita can not only promote cell proliferation, but also improve the phagocytic ability of each single RAW264.7 in vitro. A new method is provided for further study on immune activity and on the mechanism of enhancing the immune activity that some tonic herbs may have, such as D. opposita.
ABSTRACT
This article was aimed to study the chemical constituents of Dioscorea opposita Thunb.The chemical constituents were isolated and purified by Diaion HP-20,Toyopearl HW-40,Sephadex LH-20,MCI Gel CHP-20,silica gel column chromatography and preparative HPLC,TLC,purification and isolation from Dioscorea opposita Thunb.The structures of isolated compounds were identified by thc physicochemical properties and spectral analysis.The result showed that 14 compounds were isolated from Dioscorea opposita Thunb.The chemical structures were elucidated as L-Tryptophane (1),Seguinosides F (2),1-methoxycarbonyl-β-carboline (3),Helichrysin A (4),Bungein A (5),Hydroquinone (6),Zarzissine (7),Cyclo-(Pro-Thr) (8),4-Hydroxy-3-methoxybenzyl alcohol (9),pyridine-3-carboxamide (10),Arbutin (11),Methyl syringate 4-O-β-D-glucopyranoside (12),L-Phenylalanine (13),1,2-benzenedicarboxylic acid,1,2-bis[2-(2-hydroxyethoxy) ethyl] ester (14).It was concluded that chemical compounds 1-14 were isolated for the first time from Dioscorea opposita Thunb.
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In order to provide a reference for the future development, the development status of Dioscoreae Rhizoma (also called Chinese yam) has been analyzed in the view of invention patent. We made the use of Espacenet and World Traditional Medicine Patent Database to collect patent information at first. Then we performed a series of process on the collected dataset, including screening, sorting, statistics, analysis, and visualization, with the help of Python, R, SQLite, and other tools. We got the development status information of Chinese yam that reflected in patent data, involving situation, trends, international structure, function preferences, and compatibility preferences. In the perspective of patents, Chinese yam is currently in the early stages of development-term, the development space is huge for future. Chinese mainland has invested a lot in the research of Chinese yam, comparing with other regions in the world. Chinese mainland owns the major part of international patent applications. The U.S. is the most attractive region for the applicants. Chinese yam is usually used in the treatment of digestive diseases, metabolic diseases, and systemic protection or anti-depress, detailed use includes helping digestion, treatment of hyperglycemia, and enhancing immunity. The ideal in common compatibility herbs is Gan-Wen-Yi-Qi, Bu-Pi-Yi-Qi, and others.
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This study was aimed to analyze the influence of specific clinical effect of Dioscorea opposita Thunb. by dosage in order to summarize its rules. Recordings on pills containing D. opposita Thunb. from the classic prescrip-tion of the Formula Dictionary of Traditional Chinese Medicine were collected. Dosage of D. opposita Thunb. was used as the cardinal line. EXcellwas used to conduct schematic analysis. The results showed that small amount of D. opposita Thunb. had the effect of reinforcing the spleen. The effect of tonifying the lung had the dosage boundary line of 10 g. The small dosage had weak effect and large dosage had strong effect. The effect of tonifying the kidney was often the middle dosage. When the dose was larger or smaller, the effect was not obvious. The effect of consoli-dating the kidney was obvious when using larger or smaller dosage. It was concluded that the dosage of D. opposita Thunb. had relatively big influence on its effect, which provided reference of more value for the clinical practice and health cultivation.
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This article was aimed to study on clinical application of Shanyao (Dioscorea opposita Thunb.) in each historical period, which can offer some suggestions for clinical doctors to use Shanyao correctly. A total of 668 pieces of formulae containing Shanyao were recorded from the classic format Dictionary of Traditional Chinese Medicine Formula. And then, statistic analysis was made on the emerging frequency according to its effect. The results showed that different effects of Shanyao started to be widely applied from the Song dynasty. It was most widely applied in the Ming and Qing dynasty. Nowadays, the difference between using Shanyao has been gradually decreased. During the whole historical period, clinical functions of Shanyao are mainly reinforcing the spleen and the stomach, tonifying the kidney and strengthening yin. However, clinical functions of reinforcing the kidney and restraining yin, tonifying the lung and promoting fluid are used secondly. It was concluded that the research analyzed clinical application of Shanyao in each historical period, which can offer scientific basis for clinical practice and health maintenance with the using of Shanyao.
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This study was aimed to analyze the influence of Dioscorea opposita Thunb. by processing methods from the aspect of formula. The search was made on formulae containing Dioscorea opposita Thunb. from the classic book For-mula Dictionary of Traditional Chinese Medicine. Processing methods were analyzed and shown with charts by EXCEL. The results showed that the most frequently used processing methods for Dioscorea opposita Thunb. were stir-heating, dry and crude in the clinical practice. Among them, the stir-heating Dioscorea opposita Thunb. was good at reinforcing the spleen and the kidneys. The crude produce was good at tonifying the lungs. And the dry product was good at rein-forcing the kidneys. It was concluded that the processing methods had obvious influence on the clinical function of Dioscorea opposita Thunb., which can be used as reference of more value for the clinics and health cultivation.
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This study was aimed to carry out the research by statistics to analyze the influence of specific clinical function of Dioscorea opposita Thunb. from the viewpoint of prescription in order to summarize its rules. Formula containing D. opposita Thunb. in the classic book of the Formula Dictionary of Traditional Chinese Medicine were recorded. The prescription type of D. opposita Thunb. was taken as the mainline. Excellwas used in the chart analy-sis. The results showed that for the function of reinforcing spleen-stomach, the effect of powder was increased, but the effect of pill was reduced. For the function of reinforcing lung and generating essence, the effect of decoction was significantly increased, but the effect of pill was significantly reduced. For the function of reinforcing kidney-yin, the effect of pill was significantly increased, but the effect of decoction was reduced. For the function of strengthening the kidney and restraining yin, the effect of powder was reduced. It was concluded that preparation had a strong in-fluence on the clinical function of D. opposita Thunb., which provided references for both clinical practice and health cultivation.
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Objective To study the cryopreservation technique of Dioscorea opposita germplasm resources by vitrification. Methods Stems with buds of B# D. opposita germplasm were used as materials for cryopreservation by vitrification and the survival rates were examined by culture method in order to optimize a cryopresevation by vitrification technical system for D. opposita germplasm. Results A best procedure of cryopresevation by vitrification was as below: At first, the aseptic plantlets of stems with buds of B# D. opposita subcultured in vitro for 60 d were treated at 4 ℃ for 7 d. The stems with buds of D opposita about 1-1.5 cm in length were cut and precultured at 4 ℃ for 2 d in 5% sucrose and 3% mannose media, they were dehydrated with 60% vitrification solution (PVS_1: 22% glycerol+13% glycol+13% polyethyleneglycol+10% dimethyl sulfoxide) at 0 ℃ for 60 min and then dehydrated with 100% vitrification solution at 0 ℃ for 60 min. At last, the stems were immersed immediately into liquid nitrogen directly and conserved for 24 h. After rapidly thawing in a water bath at 37 ℃, the stems were washed four times with MS medium supplemented with 7% sucrose and 10 min each time, then transferred and re-cultured on the MS medium supplemented with KT 2 mg/L and NAA 0.02 mg/L. The survival rate was up to 75%. The regenerated plantlets of D. opposita showed no more difference than the normal plantlets in morphology. Conclusion This test sets up successfully a cryopresevation by vitrification system technique for D. opposita germplasm, which provides an effective way for long-lasting conservation in vitro of D. opposita germplasm resources.