ABSTRACT
The powder modification technology was used to improve the powder properties and microstructure of Dioscoreae Rhizoma extract powder, thereby solving the problem of poor solubility of Dioscoreae Rhizoma formula granules. The influence of modifier dosage and grinding time on the solubility of Dioscoreae Rhizoma extract powder was investigated with the solubility as the evaluation index, and the optimal modification process was selected. The particle size, fluidity, specific surface area, and other powder properties of Dioscoreae Rhizoma extract powder before and after modification were compared. At the same time, the changes in the microstructure before and after modification was observed by scanning electron microscope, and the modification principle was explored by combining with multi-light scatterer. The results showed that after adding lactose for powder modification, the solubility of Dioscoreae Rhizoma extract powder was significantly improved. The volume of insoluble substance in the liquid of modified Dioscoreae Rhizoma extract powder obtained by the optimal modification process was reduced from 3.8 mL to 0 mL, and the particles obtained by dry granulation of the modified powder could be completely dissolved within 2 min after being exposed to water, without affecting the content of its indicator components adenosine and allantoin. After modification, the particle size of Dioscoreae Rhizoma extract powder decreased significantly, d_(0.9) decreased from(77.55±4.57) μm to(37.91±0.42) μm, the specific surface area and porosity increased, and the hydrophilicity improved. The main mechanism of improving the solubility of Dioscoreae Rhizoma formula granules was the destruction of the "coating membrane" structure on the surface of starch granules and the dispersion of water-soluble excipients. This study introduced powder modification technology to solve the solubility problem of Dioscoreae Rhizoma formula granules, which provided data support for the improvement of product quality and technical references for the improvement of solubility of other similar varieties.
Subject(s)
Powders , Solubility , Technology, Pharmaceutical , Technology , Plant Extracts , Particle SizeABSTRACT
Dioscoreae Rhizoma formula granules are made from decoction pieces by decocting， extracting， separating， concentrating， drying and granulating， which have the advantages of simple dispensing， convenient use and easy to take without decoction. However， because Dioscoreae Rhizoma is rich in starch and mucus components， its extract powder and formula granules are poorly soluble and difficult to dissolve or disperse completely within 5 min， and the insoluble material is difficult to dissolve completely even after 24 h in water， which affects the quality evaluation of the formula granules and medication psychology of patients. Therefore， by studying the dissolution process and mechanism of Dioscoreae Rhizoma extract and its formula granules， it was found that the special chemical composition of Dioscoreae Rhizoma， the denaturation of starch and its compounding with protein and other substances during the high temperature extraction process， and the contraction of coating membrane during the spray drying process were combined to form the special microstructure of coating membrane covering starch granules， and it is the root cause of poor solubility of Dioscoreae Rhizoma formula granules. Based on the research on the structure， property and function of the powder， this paper proposed a technical strategy to improve the solubility of Dioscoreae Rhizoma formula granules by powder modification process， and experimentally demonstrated that the modified Dioscoreae Rhizoma formula granules could completely dissolve within 2 min， which solved the technical problem and could provide reference for the improvement of solubility of other similar varieties， and promote the high-quality development of traditional Chinese medicine formula granule industry.
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Objective:To investigate the mechanism of joint cavity injection of Dioscoreae Rhizoma polysaccharides （DRP） in protecting against cartilage degeneration and inhibiting the expression of inflammatory factors in the rabbit model of knee osteoarthritis to provide relevant references for the development and further research on DRP. Method:Fifty-five New Zealand white rabbits were selected for the induction of knee osteoarthritis model by the modified Hulth's modeling method. The model rabbits were randomly divided into a model group， a sodium hyaluronate group （1.00 mg·kg<sup>-1</sup>）， and low- （0.7 mg·kg<sup>-1</sup>）， medium- （1.43 mg·kg<sup>-1</sup>）， and high-dose （2.15 mg·kg<sup>-1</sup>） DRP group according to a random number table. One week after modeling， the rabbits in the groups with drug intervention were treated correspondingly for five weeks， once per week， and no intervention was performed in the model group. Five weeks later， the joint specimens were observed by visual observation. The articular cartilage tissues were observed under the light microscope for pathological sections and scores by hematoxylin-eosin （HE） staining and toluidine blue （TB） staining. The expression levels of interleukin-6 （IL-6）， interleukin-1<italic>β</italic> （IL-1<italic>β</italic>）， and tumor necrosis factor-<italic>α </italic>（TNF-<italic>α</italic>） in the synovial fluid were measured by enzyme-linked immunosorbent assay （ELISA）， and the expression of matrix metalloproteinase-13 （MMP-13）， transforming growth factor-<italic>β</italic><sub>1</sub> （TGF-<italic>β</italic><sub>1</sub>）， and type Ⅱ collagen （Col-Ⅱ） in the articular cartilage were measured by immunohistochemistry. Result:After five weeks of DRP intervention， compared with the model group， the DRP groups exhibited lowered levels of IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in the synovial fluid （<italic>P</italic><0.05）， reduced expression of MMP-13 in the articular cartilage （<italic>P</italic><0.05）， and increased levels of TGF-<italic>β</italic><sub>1 </sub>and Col-Ⅱ （<italic>P</italic><0.05）. Compared with the low-dose and high-dose DRP groups， the medium-dose DRP group showed reduced levels of IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in the knee joint （<italic>P</italic><0.05）， increased levels of TGF-<italic>β</italic><sub>1</sub> and Col-Ⅱ in cartilage tissues （<italic>P</italic><0.05）， and dwindled level of MMP-13 （<italic>P</italic><0.05）. Compared with the sodium hyaluronate group， the medium-dose DRP group showed no significant differences in IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in rabbit knee joints， and TGF-<italic>β</italic><sub>1</sub>， Col-Ⅱ， and MMP-13 in cartilage tissues. Conclusion:Joint cavity injection of DRP can significantly reduce the expression of IL-6， IL-1<italic>β</italic>， and TNF-<italic>α</italic> in rabbit synovial fluid， effectively inhibit the expression of MMP-13 in the articular cartilage to suppress the degradation of articular cartilage collagen and promote the synthesis of TGF-<italic>β</italic><sub>1</sub> and Col-Ⅱ. Therefore， DRP can protect and repair articular cartilage to delay the degeneration of articular cartilage.
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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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Objective:To study the effect and underlying mechanism of sulfur-fumigation and water-soaking on total ash of Di-oscoreae Rhizoma, find the key factor( s) affecting the total ash of Dioscoreae Rhizoma, and explore the rationality of ash limits of Di-oscoreae Rhizoma described in Chinese Pharmacopoeia. Methods:Dioscoreae Rhizoma was respectively dealt with sulfur-fumigation and water-soaking. The changes in total ash content of Dioscoreae Rhizoma was detected by the ash determination methods for total ash and SO2 described in the pharmacopoeia, and then the ash content change of inorganic salts was used to study the mechanism. Results:Sulfur-fumigation could slightly reduce the total ash content of Dioscoreae Rhizoma, while significantly reduce the ash content of calcium oxalate and calcium sulfate with the reduction degree of 7. 20% and 9. 90%, respectively. Calcium phosphate and calcium chloride were slightly affected by sulfur-fumigation, and the results indicated that the effect of sulfur-fumigation on ash content was mainly real-ized by increasing the decomposition rate of calcium oxalate and calcium sulfate. Water-soaking could decline the ash content of Di-oscoreae Rhizoma, and the phenomenon was common in the rhizome medicinal materials. The influence of water-soaking on total ash was more significant than that of sulfur fumigation. Conclusion:Sulfur-fumigation can reduce the total ash content of Dioscoreae Rhizo-ma by increasing the decomposition rate of calcium oxalate and calcium sulfate, however, the effect is mild and the process isn't the key influencing factor in the total ash content of Dioscoreae Rhizoma. During the preparation of Dioscoreae Rhizoma medicinal slices, water-soaking can cause the great loss of water-soluble mineral salts, such as Cl-, C2 O4 2-, NO3 - and SO4 2-, which leads to the reduction of total ash content, therefore, water-soaking is the key influencing factor in the total ash content of Dioscoreae Rhizoma.
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Objective: To establish an HPLC-MS/MS method for determination of allantoin in Dioscoreae Rhizoma. Methods: Determination of LC system used Agilent Eclipse Plus C18 (100 mm × 4.6 mm, 3.5 μm) column and acetonitrile-water (90:10) as mobile phase in linear gradient condition. The flow rate was 0.3 mL/min. The mass spectrometer was operated under the negative ion mode with ESI source. MRM (precursor ion of m/z 156.9 and product ion of m/z 96.7) was used to qualify allantoin. Results: The linear range of the component was 0.12-6.0 μg/mL, r=0.998 5; The average recovery was 101%, and the RSD was 3.2%. Conclusion: The method is sensitive, reliable, and accurate, and can be applied in the quality control of Dioscoreae Rhizoma.
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Objective: To provide the scientific evidence for molecular identification and phylogenetic evolution by analyzing internal transcribed spacer (ITS) sequences in 14 different germplasms from Dioscoreae Rhizoma. Methods: The ITS regions were cloned by PCR amplification and sequenced bi-directionally. The ITS sequences were aligned using Clustal X software (version 1.83), the genetic distances were calculated using Mega software (version 4.1), and the phylogenetic trees were constructed through the Neighbor-joining (NJ) and maximum parsimony (MP) methods. Results: The length of ITS sequences obtained ranged from 558 to 594 bp, of which ITS1 was from 141 to 165 bp and ITS2 was from 146 to 158 bp. There were a lot of transitions and transversions in ITS sequences, and the transition/transversion ratio among 14 different germplasms was 5.347. Both in ITS1 and ITS2, 102 variable sites were further observed. The kimura 2-parameter (K2-P) genetic distance among 14 different germplasms ranged form 0 to 0.5172. The phylogenetic trees suggested that there were close genetic relationship among Dioscorea opposita, D. persimilis, and D. japonica, they grouped closely into Clade I; Furthermore, D. alata and D. fordii also shared a closer genetic relationship comprised another clade (Clade II). Conclusion: The phylogenetic analyses based on ITS sequences can present a foundation for clearing the evolution of germplasm resources in Dioscoreae Rhizoma; The variable bases can also provide reliable molecular evidences for identifying different genotypic germplasms of Dioscoreae Rhizoma.
ABSTRACT
Dioscoreae Rhizome (DR) has been used in traditional medicine to treat numerous diseases and is reported to have anti-diabetes and anti-tumor activities. To identify a bioactive traditional medicine with anti-inflammatory activity of a water extract of DR (EDR), we determined the mRNA and protein levels of proinflammatory cytokines in macrophages through RT-PCR and western blot analysis and performed a FACS analysis for measuring surface molecules. EDR dose-dependently decreased the production of NO and pro-inflammatory cytokines such as IL-1beta, IL-6, TNF-alpha, and PGE2, as well as mRNA levels of iNOS, COX-2, and pro-inflammatory cytokines, as determined by western blot and RT-PCR analysis, respectively. The expression of co-stimulatory molecules such as B7-1 and B7-2 was also reduced by EDR. Furthermore, activation of the nuclear transcription factor, NF-kappaB, but not that of IL-4 and IL-10, in macrophages was inhibited by EDR. These results show that EDR decreased pro-inflammatory cytokines via inhibition of NF-kappaB-dependent inflammatory protein level, suggesting that EDR could be a useful immunomodulatory agent for treating immunological diseases.