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OBJECTIVE: To investigate the effects of different processing methods on antitussive and hepatotoxicity of Dioscorea bulbifera L.. METHODS The raw and processed products of Dioscorea bulbifera L., fried Dioscorea bulbifera L. (FD), wine-fried Dioscorea bulbifera L. (WD), liquorice liquid-fried Dioscorea bulbifera L. (LD) and angelicae liquid-fried Dioscorea bulbifera L. (AD) were administered to mice at a dose of 1.7 g•kg-1 for 7 d. The effects of processing on the cough relief of Dioscorea bulbifera L. were evaluated by the cough test induced by concentrated ammonia water. The detoxification effect and the preliminary mechanism of processing on Dioscorea bulbifera L. were evaluated by the detection and analysis of the relevant indicators. RESULTS: Compared with the RD group, LD and AD group had longer cough latency (P0.05). Compared with the AD group, there were more coughing times in LD group (P0.05) in mice induced by concentrated ammonia water. The intervention of WD only had significant reversal effect on the low GST level of liver (P0.05). CONCLUSION: Processing with Angelica sinensis and licorice could enhance the antitussive effect of Dioscorea bulbifera L. and reduce its hepatotoxicity. The mechanism of attenuation may be related to the inhibition of lipid peroxidation and the enhancement of antioxidant level in liver. The antitussive effect of two kinds of processing, stir-fried and wine-fried, is only effective, but no synergistic effect, no toxic effect and no attenuation effect are found.
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Objective To explore the characteristics, diagnosis, treatment and prognosis of liver injury caused by the deficiency of dioscorea bulbifera L.. Methods The general data, clinical manifestation and laboratory examination of 45 cases of liver injury diagnosed as Yoshimoto associated liver injury from November 2014 to June 2017 were classified and reviewed with the standards of drug liver injury classification recommended by the Council of international medical organizations. Results The number of male patients was 26, and female 19. The medication time ranged from 1 week to 2 years and the main biochemical performance was abnormal, namely ALT, AST, TBil, DBil, ALP and GGT. Of the 45 cases, the average values of ALT, AST were 608.11 ± 411.30 U/L and 505.38 ± 342.15 U/L. The TBil of 42 case rised with the mean value 170.10 ± 136.86 μmol/L, and the ALP of 22 cases with 182.38 ± 55.15 U/L. The GGT of 43 cases rised with the mean 223.12 ± 131.85 U/L. Clinical classification included 38 cases were liver cell injury, none was cholestasis, 5 mixed types and 2 cases of liver biochemical examination abnormality. One patient died while the other patients recovered. Conclusions Although the pathogenesis of the liver cell induced injury type with dioscorea bulbifera L. remains unclear, the reasonable and appropriate use of medication and regular liver biochemical tests is necessary.
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OBJECTIVE To characterize pharmacokinetics and tissue distribution of diosbulbin B (DIOB)in rats,and compare exposure and elimination of DIOB in plasma and tissues after ig adminis-tration of compound DIOB and ethanol extract of Dioscorea bulbifera L.(EEDB),respectively.METHODS Liquid chromatography coupled with tandem mass spectrometry method was developed and validated for quantitative determination of DIOB in plasma and tissue samples. The sensitivity, matrix effect, accuracy and precision of the method were determined. After a single ig administration of DIOB 1.3 mg·kg-1and EEDB 1.0 g·kg-1, respectively, plasma and tissues samples were collected separately from male SD rats at prescheduled time points.The drug-protein binding of DIOB in plasma and tissues was measured using Rapid Equilibrium Dialysis Assay. The concentration-time profiles of DIOB were analyzed using WinNonLin 6.4 to obtain pharmacokinetic parameters. RESULTS Rapid absorption and elimination of DIOB were observed in rats.The plasma maximum concentration(cmax),peak time(tmax) and eliminate half life(t1/2)of DIOB in rats receiving DIOB or EEDB were 312±67 and(131±84)μg·L-1(P<0.01), 0.12 ± 0.07 and (0.23 ± 0.17) h, and 1.17 ± 0.28 and (2.34 ± 0.83) h (P<0.05). DIOB was rapidly distributed in most tissues,with the AUC(0-24 h)of DIOB in lungs,liver and kidneys for DIOB and EEDB groups were 4527.0±557.7,183.0±51.1 and(64.4±22.4)ng·h·g-1,and 6507.9±424.3(P<0.01),467.5±202.7(P<0.05) and (238.6 ± 70.0) ng·h·g-1(P<0.05), respectively. The protein binding rate of DIOB was 38.7%-43.6%,61.3%-66.9% and 61.4%-64.7% in plasma,livers and lungs,respectively.The ratio of free concen-trations between tissue and plasma for lungs was more than 48,indicating the specific distribution of DIOB in lungs.CONCLUSION There is difference in pharmacokinetics and tissue distribution of DIOB in rats after taking compound DIOB and EEDB.Among the tissues detected,lungs are the major target of DIOB with the highest exposure level.These characterizations should be considered in pharmacological and toxicological studies of DIOB and relevant herbs.
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Con el objetivo de conocer y rescatar la historia, usos y manejo dado a Dioscorea bulbifera L. por las comunidades campesinas del distrito de Donoso (provincia de Colón, Panamá) se llevó a cabo un estudio etnobotánico en el primer semestre de 2012. El mismo se desarrolló a partir de grupos focales, recorridos de campo y entrevistas semiestructuradas a personajes clave (ancianos, curanderos y personas que cultivan D. bulbifera). Se realizaron cinco grupos focales y 11 entrevistas semiestructuradas, abordando aspectos relativos a historia, nombres, manejo tradicional, usos y formas de consumo de la papa de aire en la región. Como resultados destacados se tiene que D. bulbifera es una especie de muy vieja presencia en las comunidades, teniendo como uso principal la alimentación humana. Igualmente, los agricultores dan cuenta de la importancia de esta especie como alimento altamente nutritivo y con algunos usos medicinales. La información recabada sugiere un conocimiento erosionado respecto de esta especie, lo cual se refleja en el nivel de conocimiento de su manejo y la disminución de su cultivo. No obstante, el conocimiento rescatado resulta de gran utilidad para el establecimiento de ensayos que se orienten a la sistematización de prácticas de cultivo de esta especie.
In order to know and rescue the history, uses and management of Dioscorea bulbifera L. given by peasant communities in the district of Donoso (Colón Province, Panamá) an ethno-botanical study was conducted in the first semester 2012. The study was developed from focus groups, field observations and semi-structured interviews to key figures (elders, healers and people who plant D. bulbifera). Five focus groups and eleven semi-structured interviews were conducted, addressing aspects of history, names, traditional management, uses and forms of consumption of air potatoes in the region. Some outstanding results, demonstrate that Dioscorea bulbifera is a species of very old presence in the communities, used mainly for human consumption. Furthermore, farmers report the importance of this species as a highly nutritious food and having some medicinal uses. The information gathered suggests an eroded knowledge about this species, which is reflected in the level of knowledge of its handling and decreased cultivation. However, the rescued knowledge is useful for establishing essays that aim to systematize practices for cultivation of this species.
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Humans , Panama , Knowledge , Dioscorea , Food SupplyABSTRACT
Objective To establish a HPLC method for determining and comparing the contents of diosbulbin B in Dioscorea bulbifera L. from different regions. Methods The medicine powders were extracted twice by water, then the water extracts were combined and detected by HPLC at 35℃,with Kromasil-C18 column (4. 6 mm×250 mm,5μm) and a mobile phase of acetonitrile-water-glacial-acetic acid (34660. 1). The flow rate was 1. 0 mL·min-1 and the detection wavelength was 210 nm. Results The linear range of diosbulbin B was 26. 0-260. 0 μg·mL-1(r=0. 999 9). The average content of diosbulbin B in Dioscorea bulbifera L. from different areas was that of Hebei>Guangxi>Jiangsu>Sichuan>Zhejiang. Conclusion The method is simple,quick,accurate and suitable for the determination of diosbulbin B in Dioscorea bulbifera L. from different regions.
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Objective: In order to provide a theoretical basis for clarifying the mechanism of synthesis of saponins during different induction formation period of Dioscorea bulbifera microtubers, squalene synthase (SQS) gene expression of D. bulbifera microtubers was analyzed in this paper. Methods: Using Actin gene as a reference gene, Real-time quantitative PCR (qRT-PCR) analysis technology was applied. The amplification curve and solubility curve of SQS (target gene) and Actin (reference gene) were successfully constructed by SYBR Green I after RNA was extracted from different formation period of D. bulbifera microtuber and cDNA was obtained by reverse transcription. Result: Quantitative results showed that: During the initial stage (18-36 d) of D. bulbifera microtuber formation, the expression level of SQS gene increased significantly. During the mid-term (36-72 d) of D. bulbifera microtuber formation, the expression level of SQS gene decreased significantly, and tended to be stable. During the later stage (72-90 d) of D. bulbifera microtuber formation, the expression level of SQS gene decreased significantly again. Conclusion: In general, in the process of D. bulbifera microtuber formation, the expression level of SQS gene shows the trend of "low-high-low-constant-low", which indicates that SQS is a key enzyme of saponin biosynthesis during the different induction formation period of D. bulbifera microtubers.
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Objective: To discuss the influence of several factors on the cryopreservation of embryogenic calli induced from Dioscorea bulbifera microtuber by droplet-vitrification and to test the genetic stability of the regenerated plantlets after freezing from the aspects of morphology, physiology, DNA content, as well as the photosynthetic characteristics and chlorophyll fluorescence parameters in this paper. Methods: Plant tissue culture (including microtuber induction and embryogenic callus induction), plant physiology index detection (including total chlorophyll, soluble protein, soluble sugar and superoxide dismutase enzyme and peroxide enzyme activity), and cell flow cytometry were applied. Results: The best cryopreservation conditions of embryogenic callus of D. bulbifera microtuber were as following: Embryogenic calli were precultured in liquid media of MS+KT 2 mg/L+NAA 0.5 mg/L+2, 4-D 0.5 mg/L+0.3 mol/L sucrose for 1 d and then treated in loading liquid (MS+2 mol/L glycerol+0.4 mol/L sucrose, pH 5.8) for 20 min. In order to dehydrate, embryogenic calli were transferred in 100% PVS2 at 0℃ for 40 min. After dehydration, the embryogenic calli were inoculated to PVS2 small drops in the aluminum foil strips and then dipped in liquid nitrogen (LN). Finally the aluminum foil strips were quickly transferred to freezing tube that filled with LN and then put into LN tank. After conserving for 1 d in LN, the aluminum foil strips were removed and the embryogenic calli were immersed into liquid washing media (MS+KT 2 mg/L+NAA 0.5 mg/L+2, 4-D 0.5 mg/L+1.2 mol/L sucrose, pH 5.8) preheated in 37℃ warm water. After separated from the aluminum foil strips, the embryogenic calli were washed with fresh liquid washing media at room temperature for there times, 10 min each time. After washing, the embryogenic calli were transferred onto differentiation medium (MS+KT 2 mg/L+NAA 0.5 mg/L+30 g/L sucrose+5 g/L agar), and cultured in dark for 2 d and then cultured in 12 h/d photoperiod, the cell survival rate reaches above 89%. The morphological and physiological indexes and the content of DNA of two kinds of plantlets, which regenerated from cryopreserved and non-cryopreserved embryogenic calli induced from D. bulbifera microtuber by droplet-vitrification, showed no significant difference (P>0.05). Conclusion: Cryopreservation technology system of embryogenic calli induced from D. bulbifera microtuber by droplet-vitrification is established and the regeneration plants have no genetic variation, which provides the theoretical basis and technical basis for the long-term preservation of germplasm resources in the plants of Dioscorea L.
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Objective: In order to find the suitable concentration and combination of plant growth regulators, the effects of plant growth regulators (NAA, 2, 4-D, 6-BA, KT, and PP333) on in vitro induction formation for the plantlet microtuber of Dioscorea bulbifera was studied. Methods: Through plant tissue culture technique, single factor test, and orthogonal test, taking the stems with a bud of D. bulbifera plantlets as explants, the effects of plant growth regulators on the in vitro induction formation for the microtubers of D. bulbifera were investigated. Results: Auxin using alone was conducive to the induction formation for the microtuber of D. bulbifera. The suitable concentration of both NAA and 2, 4-D inducing the microtuber formation was 0.5 mg/L, but the inducing effects of NAA and 2, 4-D had no significant difference. Cytokinin using alone was not conducive to the induction formation for the microtuber of D. bulbifera. The suitable concentration of both KT and 6-BA inducing microtuber formation was 2 mg/L, but the inducing effect of KT is better than that of 6-BA. The combination of auxin, cytokinin, and PP333 could significantly promote the in vitro induction formation for the microtuber of D. bulbifera, the better combination was MS+NAA 0.5 mg/L+6-BA 2.0 mg/L+PP333 0.5 mg/L. Conclusion: Based on these experimental results, the paper selects the suitable concentration of plant growth regulators conducive to the in vitro induction formation for the microtuber of D. bulbifera, which has laid the technical foundation for their in vitro induction formation of microtuber and factory production.
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Objective Effects of different factors on proliferation and rooting of stems with a bud were studied by using Dioscorea bulbifera as test material to optimize the rapid propagation system of D.bulbifera virus-free plantlets.Methods Plant tissue culture method was used in shoot tip culture and rapid propagation study,and RT-PCR method was used in virus detection of virus-free plantlets.Results The best proliferation medium of D.bulbifera stems with a bud was MS+KT 2 mg/L+6-BA 1 mg/L+NAA 0.5 mg/L;The best sucrose and agar concentration of D.bulbifera stems with a bud was 30 g/L and 0 g/L,respectively;The best rooting medium of D.bulbifera stems with a bud was 1/2 MS+IBA 0.1 mg/L+NAA 0.5 mg/L+PP_(333) 1 mg/L;The best transplanting matrix of regeneration plantlets from D. bulbifera stems with a bud was perlite-vermiculite(2:1);The best PP_(333) concentration of D.bulbifera regeneration plantlets for transplanting was 50 mg/L.Conclusion The rapid propagation system of D. bulbifera virus-free plantlets is established successfully for the first time,which provides a technological basis for factory production of D.bulbifera virus-free plantlets.
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Objective Effects of different factors on shoot tip culture in vitro were studied by using Dioscorea bulbifera as test material to find the media suitable to the shoot tip differentiation of D.bulbifera and the method of virus delection.Methods Plant tissue culture method was used in shoot tip culture and the symptom observation method,instruction plant method,and RT-PCR method were used in virus detection of virus-free plantlets.Results The best disinfection method for D.bulbifera shoot tips was firstly disinfected for 30 s with 70% alcohol and then disinfected for 12 min with 0.1% HgCl2;It is better for D.bulbifera to cut shoot tips in 0.5 mm length after 37 ℃ heat treatment for 7 d;The best proliferation medium of D.bulbifera shoot tips was MS+KT 2 mg/L+NAA 0.5 mg/L;The best rooting medium of regeneration buds from D.bulbifera shoot tips was 1/2 MS+NAA 0.5 mg/L;The best matrix of regeneration plantlets from D.bulbifera shoot tips was perlite-vermiculite (2∶1);RT-PCR Method was primary mean to detect potato virus Y (PVY) of regeneration plantlets from D.bulbifera shoot tips,and average rate of PVY-free was 86.5% by RT-PCR.Conclusion The virus-free culture system of D.bulbifera shoot tips is established for the first time,providing a technological basis for the rapid propagation and factory production of D.bulbifera virus-free plantlets.
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Objective To study the effects of several factors on bud proliferation and rooting of Dioscorea bulbifera stem with a bud.Methods Single factor test and plant tissue culture methods were applied.Results The combination of 6-BA or KT and NAA helped the proliferation of stem with a bud in D.bulbifera;high concentration sucrose led to callus growth of stem with a bud in D.bulbifera which didn′t help the proliferation of stem with a bud in D.bulbifera;Liquid culture also helped the proliferation of stem with a bud in D.bulbifera;In a certain range,the increase of NAA concentration helped the rooting of stem with a bud in D.bulbifera.But it will inhibit the rooting with concentration in a higher level.Conclusion The best proliferation culture medium of D.bulbifera,stem with a bud is MS + KT 2 mg/L +NAA 0.5 mg/L or MS + 6-BA 2 mg/L + NAA 0.5 mg/L;For proliferation of D.bulbifera stem with a bud,the best sucrose concentration is 30 g/L sucrose,the best culture mode is liquid culture;The best rooting culture medium of D.bulbifera stem with a bud is MS + NAA 2 mg/L.