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Objective To explore the antitumor small molecules targeting the ubiquitin–proteasome system (UPS) on the basis of active molecules from traditional Chinese medicine. Methods UbG76V-GFP stably expressing cell line was constructed to screen novel small molecule inhibitors targeting UPS. The fluorogenic substrates of Suc-LLVY-AMC, Z-LLE-AMC, and Boc-LRR-AMC were used to assess the effect of dioscin on the 20S proteasome hydrolase activity. The Ub-AMC substrate was used to evaluate the effect of dioscin on the intracellular deubiquitinating enzyme activity. Western blot was used to detect the effect of dioscin on intracellular ubiquitination levels. CCK-8 and colony formation assays were used to detect the inhibitory effect of dioscin on the tumor cell proliferation. Results Dioscin is a UPS inhibitor discovered through the UbG76V-GFP reporter system. It enhances intracellular ubiquitination and inhibits tumor cell proliferation and colony formation by targeting deubiquitinating enzymes. Conclusion Dioscin could significantly inhibit tumor cell proliferation by targeting ubiquitin–proteasome.
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AIM: To observe the effect of Dioscin treatment on NF-κB signaling pathway and cellular inflammatory injury and explore its potential mechanism in uric acid-induced mouse tubular epithelial cells (mTECs). METHODS: After 1.2 mol/L uric acid induced mTECs, Dioscin and NF-κB P65 inhibitor BAY11-7082 were given to intervene respectively. IκB-α, NF-κB P65, PP65, NLRP3, IL-1β and β-actin were detected by Western Blot, immunofluorescence staining and real-time PCR. RESULTS: Western Blot, immunofluorescence staining and real-time PCR analysis showed that expression levels of PP65, NLRP3 and IL-1β were significantly downregulated in the uric acid-induced mTECs with Dioscin and BAY11-7082 treatment. CONCLUSION: Dioscin attenuates uric acid-induced cellular inflammatory damage by suppression NF-κB signaling pathway.
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Aim To investigate the effects and possible molecular mechanisms of dioscin(DIO)against depression in mice.Methods Eighty mice were randomly divided into control group, DIO control group, model group, DIO groups(20, 40 and 80 mg·kg-1 DIO)and FLU group(20 mg·kg-1 fluoxetine).After establishing the depression model with chronic unpredictable moderate stress(CUMS)in mice, the corresponding drugs were administered by gavage continuously for four weeks in each group.The behavior of mice was tested, and the contents of corticosterone(CORT), brain-derived neurotrophic factor(BDNF), 5-hydroxytryptamine(5-HT), malondialdehyde(MDA), superoxide dismutase(SOD)and catalase(CAT)were evaluated by ELISA or enzyme labeling method.In addition, HE staining, Nissl staining and PET scanning were operated for the brain tissues.Western blot was used to detect the protein expressions.Results Compared with model group, DIO significantly improved the behaviors of depressed mice.And it reduced the contents of CORT in serum, increased BDNF and 5-HT in hippocampus.Meanwhile, DIO obviously reduced MDA in serum, increased SOD in serum and CAT in brain tissues.DIO improved the steatosis of brain tissues, disorder and looseness of neurons, and increased glucose metabolism in brain tissues of depressed mice.The molecular mechanism suggested that compared with model group, DIO significantly increased the protein level of UCP2 to adjust the levels of Nrf2, SOD2, GLUT1 and G6Pase.Conclusions DIO improves the depression symptoms of depressed mice, which should be through adjusting UCP2-mediated oxidative stress and glucose metabolism.
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OBJECTIVE：To prepare GGPFV-modified Daunorubicin/dioscin liposomes ，and to optimize their formulation and to preliminarily evaluate their cytotoxicity to breast cancer cells in vitro . METHODS ：Daunorubicin and diosgenin were wrapped by thin film dispersion method and ammonium sulfate hydration method ；the surface was modified with DSPE-PEG2000-GGPFV to prepare GGPFV-modified Daunorubicin/dioscin liposomes. Taking encapsulation rate as index ，Box-Behnken response surface methodology was used to optimize the film hydration volume ，cholesterol amount and daunorubicin amount in the formulation. The entrapment efficiency of 3 batches of liposomes prepared according to the optimal formulation was determined. The effects of Daunorubicin/dioscin liposomes ，GGPFV-modified Daunorubicin/dioscin liposomes and blank liposomes on the survival rate of human breast cancer MDA-MB- 435S cells were compared. RESULTS ：The optimal formulation was as film hydration volume of 5 mL，cholesterol of 4 mg，yolk lecithin of 22 mg，daunorubicin of 0.55 mg，dioscin of 0.85 mg，DSPE-PEG2000 of 3.5 mg， DSPE-PEG2000-GGPFV of 2 mg. The encapsulation rate of daunorubicin was （96.21±1.54）％ and that of dioscin was （95.39± 2.48）％ in the 3 batches of liposomes prepared. The in vitro cytotoxicity tests showed that the inhibition effect of GGPFV-modified Daunorubicin/dioscin liposome on MDA-MB-435S cells was significantly stronger than that of Daunorubicin/dioscin liposome （P＜ 0.05）. There was no cytotoxicity in the membrane. CONCLUSIONS ：GGPFV-modified Daunorubicin/dioscin liposomes are successfully prepared ，and its inhibitory effect on human breast cancer MDA-MB- 435S cells in vitro was significantly enhanced.
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In this report, a heat and high-pressure homogenization method was used to prepare dioscin nanostructured lipid carriers, and the formulation of dioscin nanostructured lipid carriers was optimized by central composite design-response surface methodology. In vitro evaluation data showed that the preparation of dioscin nanostructured lipid carriers under optimal process by central composite design-response surface methodology had a spherical shape and homogeneous size distribution, with a particle size of (90.9±0.6) nm, a polydispersity index of (0.253±0.07), Zeta potential of (-45.7±0.5) mV, encapsulation efficiency of (90.2±0.5)%, and the drug loading of (23.30±0.10)%. These results clearly indicate that the preparation of dioscin nanostructured lipid carriers made with the heat and high-pressure homogenization method have very good physical and chemical properties, suitable for therapeutic applications.
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Dioscin is the main ingredient of Dioscorea,a tradi-tional medical herb.Traditional theory of Chinese medicine be-lieves that Dioscorea has effects of clearing lung,digesting food, diuresis, improving blood circulation, relaxing muscles and stopping attack of malaria.Pharmacological studies have shown that dioscin has many pharmacological effects,particularly the anti-tumor effect.Many studies have also shown dioscin im-proves symptoms of atherosclerosis and protects blood vessel en-dothelium,reduces ischemia-reperfusion injury of heart,brain and kidney,lowers blood sugar,inhibits hepatic fibrosis,im-proves menopausal osteoporosis,relieves rheumatoid arthritis, ulcerative colitis and other inflammatory disorders,and posses-ses anti-bacterial and anti-viral activity.This article focuses on the progress of the modern pharmacological study of dioscin,and reports its advances in recent years.
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Objective To study absorption characteristic of dioscin from Dioscorea nipponica Makino extract in rat intestine.Methods Single-pass intestinal perfusion (SPIP) model was used for rat in situ and HPLC was used to determine the concentrations of dioscin.The effects of different intestinal segments,drug concentration and P-glycoprotein (P-gp) inhibitor on intestinal absorption were investigated.Results Dioscin could be absorbed in the whole intestine,the absorption rate constant (Ka) and the apparent coefficient (Papp) of dioscin decreased following the sequence of ileum > duodenum =jejunum > colon.Absorption parameters of dioscin had no significant difference at different concentrations (40,80,120 mg·L-1).There were significant differences in Ka and Papp values between P-gp inhibitor group and no P-gp inhibitor group(P<0.05).Conclusion The saturate phenomena was not observed under the test range of drug concentration,and the absorption mechanism may be passive diffusion transport.Dioscin in Dioscorea nipponica Makino extract may be the substrate of P-gp.
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Objective: To develop an HPLC-DVD wavelength switching combined with gradient elution method for the determination of the contents of hydroxysafflor yellow A, protodioscin, dioscin, methylprotodioscin, pseudoprotodioscin, and gracillin in Dieda Zhitong San (DZS) simultaneously. Methods: The chromatographic separation was achieved on a Diamonsil C18 (250 mm × 4.6 mm, 5 μm) with acetonitrile (A)-0.4% formic acid solution (B) as mobile phase at the flow rate of 0.9 mL/min for gradient elution; Hydroxysafflor yellow A was detected at 403 nm; Protodioscin, dioscin, methylprotodioscin, pseudoprotodioscin, and gracillin were detected at 203 nm; Sample quantity was 20 μL. Results: The six active components were well separated and showed good linearity, such as hydroxysafflor yellow A 3.46-69.20 μg/mL (r = 0.999 4), protodioscin 9.52-190.40 μg/mL (r = 0.999 7), dioscin 8.74-174.80 μg/mL (r = 0.999 6), methylprotodioscin 4.45-89.00 μg/mL (r = 0.999 9), pseudoprotodioscin 2.64-52.80 μg/mL (r = 0.999 5), and gracillin 3.28-65.60 μg/mL (r = 0.999 8). The precision and repeatability were good, and RSD values were less than 2.0%. The stability was good in 12 h. The average recoveries and the corresponding RSD values were 98.57% (1.59%), 97.64% (1.28%), 99.43% (1.07%), 97.98% (1.64%), 98.57% (1.16%), and 97.17% (1.37%), respectively. Conclusion: An HPLC wavelength switching combined with gradient elution method has been successfully established for simultaneous determination of six components in DZS. The method is simple, quick, accurate, and helpful for the quality control of DZS.
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Dioscin chemical compositions are the main effective components in clinical commonly used Chinese medicines such as Diaoxinxuekang capsules and Xinnaoshutong capsules etc , which show distinct curative effect on cardiovascular and cerebrovascular diseases.Meanwhile, they are the important raw materials for the synthesis of steroid hormone drugs .The studies on the extraction technology exhibit important significance in the exploration of pharmacological activities of the components , which also are the external requirements for the growing demand of steroid hormone drugs market .In this paper , the relatively mature extraction methods re-searched in recent years were summarized ,and the advantages and disadvantages of the different processes were discussed in order to provide reference for the further studies and application .
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Aim To investigate the effects of dioscin ( Dio) on rat myocardial contractility. Methods Left ventricular contractile function was measured using the Langendorff non-recirculating mode of isolated rat heart perfusion. Effects of low, middle and high concentra-tion of Dio were investigated by measuring left ventricu-lar systolic pressure ( LVSP ) and left ventricular end diastolic pressure ( LVEDP) . Also, peak rates of rise/fall of left ventricular pressure ( ± dp/dtmax ) of isolated rat heart were calculated. Effects of Dio on intracellu-lar free calcium concentration in rat H9 c2 cells were measured by using the confocal microscopy. Mitochon-drial membrane potential was detected with multifunc-tional microplate reader. Results With 0. 1, 1 μmol · L-1 Dio, LVSP were significantly enhanced from (11. 55 ± 0. 52), (10. 53 ± 0. 28) kPa to (13. 08 ± 0. 72), (12. 53 ±0. 64) kPa(P<0. 01); +dp/dtmax were dramatically increased from ( 0. 38 ± 0. 10 ) , (0. 40 ± 0. 07) kPa·ms-1 to (0. 42 ± 0. 11), (0. 43 ± 0. 02) kPa·ms-1(P<0. 05). With the 10μmol· L-1 Dio, LVSP and + dp/dtmax were both decreased from (12. 13 ± 0. 33) kPa and (0. 42 ± 0. 04) kPa· ms-1 to ( 9. 46 ± 0. 77 ) kPa and ( 0. 24 ± 0. 04 ) kPa ·ms-1 (P <0. 01). With 0. 1, 1, 10 μmol·L-1 Dio, the relative fluorescence intensity of intracellular free calcium concentrations was increased significantly from (16. 62 ± 0. 89) to (21. 48 ± 0. 80), (25. 68 ± 0. 69) and (19. 84 ± 0. 66)(P <0. 01)respectively. 0. 1, 1μmol·L-1 Dio showed no significant effects on the mitochondrial membrane potential of rat H9 c2 cells, while with effects of 10 μmol·L-1 Dio, the ra-tio of JC-1 monomer and J-aggregates was changed from (1. 14 ± 0. 03) to (1. 35 ± 0. 06)(P<0. 01), indica-ting a decrease in the mitochondrial membrane poten-tial. Conclusion Low and middle concentrations of Dio show a positive inotropic effect on isolated rat heart, as the LVSP and + dp/dtmax are enhanced, which may concern with the increase of the intracellu-lar concentration of Ca2+. It will not cause the calcium overload while the intracellular concentration of Ca2+ is increased by low and middle concentration of Dio in the myocytes except high concentration of Dio.
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Dioscin is the main raw material for the synthesis of steroid hormone drugs. Currently, the direct acid hydrolysis is the mainly industrial production method for dioscin. However, the use of strong acid can not only destroy the structure of dioscin resulting in very low yield, but also produce a large amount of waste water and residues, which seriously pollute the environment. So the clean production of dioscin is the urgent demand of water conservation and environmental protection. In the paper, the recent research pro-gresses in the clean production technology and process of dioscin were summarized, and the advantages and problems were discussed in order to provide reference for the improvement and application of the new technology and process.
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Objective: To investigate the effects of serum containing dioscin on sodium current (INa) and reveal the mechanisms of cardioprotection and antiarrhymias. Methods: Serum was obtained from the aortaventralis from Wistar rats after ig administration 4 d of dioscin 300 mg/kg, twice daily for 4 d. Single ventricular myocytes were isolated from adult rat hearts by enzymatic dissociation and the effects of dioscin on INa were observed by whole-cell patch clamp. Results: Serum containing dioscin shifted downward the I-V curve with increased peak current density. With the effects of 1, 10, and 100 μL serum containing dioscin, the peak current density was dose-dependently changed from (-52.10 ± 3.80) pA/pF to (-76.44 ± 4.09) pA/pF and (-81.96 ± 4.70) pA/pF, respectively (P < 0.01 vs control); Dioscin facilitated the activation process with the inactivation process unchanged. The half activation potential was changed from (-57.69 ± 1.86) mV to (-59.71 ± 2.57) mV, ( -66.56 ± 1.32) mV (P < 0.01 vs control), and (-68.52 ± 3.91) mV (P < 0.01 vs control), respectively; The recovery process of sodium channel was accelerated by 1, 10, and 100 μL serum containing dioscin with the recovery constant τ changed from (150.73 ± 21.49) ms to (143.19 ± 13.88) ms, (84.83 ± 18.03) ms (P < 0.01 vs control), and (80.63 ± 13.89) ms (P < 0.01 vs control). Conclusion: Serum containing dioscin could increase the sodium current by facilitating the activation process and accelerating the recovery process of sodium channel.
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OBJECTIVE: To discuss the mechanism of dioscin against apoptosis of cardiomyocytes induced by hydrogen peroxide through Serum Pharmacologic Method. METHODS: The drug-containing serum [0.6 g (crude drug) · kg-1] was prepared by serum pharmacologic method. Cardiomyocytes from neonatal SD rats were cultured in Dulbecco Modified Eagle Medium (DMEM). The primary cultured cardiomyocytes were injured by hydrogen peroxide before giving the drug-containing serum with different concentrations. The cardiomyocyte viability was determined by MTT method. Morphological changes of cardiomyocytes were observed by fluorescence microscope after treating with the drug-containing serum at different concentrations. The anti-apoptotic effect of dioscin was indicated by detecting the activity of caspase-3. The protein expression of Bcl-2 and Bax were semi-quantified by Western-blot method after treating with the drug-containing serum. RESULTS: The cardiomyocyte viability was elevated (P < 0.01) after being treated with different concentrates of drug-containing serum (0.4, 0.8, 1.2 g (crude drug) · kg-1, prepared from the original drug-containing serum). Typical apoptotic features of the cardiomyocytes such as membrane blebbing, cell shrinkage and detachment, and nucleus condensation and fragmentation were dose-dependently improved after being treated with drug-containing serum. The activity of caspase-3 decreased with the increasing concentration of drug. Western blot approach showed that the Bcl-2 level increased meanwhile the Bax decreased. CONCLUSION: Dioscin could increase the viability of the primary cultured cardiomyocytes in hydrogen peroxide induced injury. It could also decrease the activity of caspase-3, improve nucleus condensation and fragmentation, increase Bcl-2 level and decrease the Bax level. Copyright 2012 by the Chinese Pharmaceutical Association.
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Objective To control the quality of the species in Dioscorea L. better. Methods An HPLC-ELSD method was developed for the first time to simultaneously determine four bioactive ingredients:dioscin gracillin,protoneodioscin, and protoneogracillin in 31 samples belonging to seven species of Dioscorea L. from different areas. The column was an Inertsil HILIC (250mmx4.6 mm,5pm). The separation was carried out with a gradient program. The mobile phase was acetonitrile-water at a flow rate of 0.8 mL/min. Results The standard curve was rectilinear in the range of 0.464-12.97 gg (r=0.9969) for dioscin, 0.310-7.09 ltg (r = 0.9953) for gracillin, 0.469-11.66 gg (r=0.9970) for protoneodioscin, and 0.276-6.87 gg (r＝0.9992) for protoneogracillin. The recoveries of the markers were 98.1%, 100.1%, 97.2%, and 96.4%, respectively. The contents of the four components were quite different among the seven species of Dioscorea L. Conclusion The proposed HPLC-ELSD method is convenient,fast, accurate, and applicable for simultaneous analysis of multiple bioactive components of species in Dioscorea L.for quality control, which could facilitate discovering new natural resources of steroidal saponin.
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OBJECTIVE:To isolate and identify the major constituents-steroidal saponins from Di'ao xinxuekang.METHODS:The constituents were separated and purified using normal-pressured and pressurized silica gel column chromatography technology,and the structures were identified based on the physico-chemical property and spectral analysis.RESULTS:Two steroidal saponins were obtained from Di'ao xinxuekang and their structures were identified as dioscin(1)and pseudoprodioscin(2).CONCLUSION:Compound 1 and 2 were demonstrated to be two major constituents in Di'ao xinxuekang.This finding is of great importance for the quality control and evaluation of Di'ao xinxuekang(Chinese Medicine).
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Objective:To discuss the regulation of berberine and dioscin extracted from traditional Chinese medicine in the expression of protein moleculer related with glucose metabolism in trophoblast cells,such as IRS-1,P1-3K,Glut1,PPAR ?. Methods:To culture the chorion trophoblast cell in the early pregnancy,to induce cells to suffer glucose metabolism obstacle with the use of WortmaninnTake advantage of berberine and dioscin extracted from the Chinese traditional medicine with intervention,simultaneously set the troglitazone(T) and dimthyl(R) for the control group. Detect the gene expression with the use of RT-PCT,simutaniously detect the protein expression in molecular level. Examine the expression in the protein level with the technique of Western blot in combination with the technique of LSM for the expression location of protein moleculer related.Results:① With the induction of WT,the glucose metabolism inside the tropholast cells becomes abnormal,compared with normal(P
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ObjectTo study the adsorptive properties of spherical activated carbon on some effective ingredients from Chinese materia medica (CMM) and to approach the application feasibility to CMM purification. Methods The spherical activated carbon with high strength has been successfully prepared by the carbonation and activation of polymeric resin. The apparent and theoretical adsorption capacities under the condition of static operation were investigated by Langmuir monolayer adsorption model. Elution by 75% alcohol was studied. Results The static adsorption capacities of the berberine hydrochloride and dioscin were 35.46 mg/g and 47.12 mg/g, and theoretical adsorption capacities were 96.16 mg/g and 102.04 mg/g, respectively. The apparent adsorption amount of rutin was 40.88% mg/g. The elution ratio were 83.71%, 91.45%, and 87.69%, respectively. Conclusion The spherical activated carbon adsorbent shows better comprehensive adsorption properties and can be used to purification.
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OBJECTIVE:To study the in vitro and in vivo metabolism of DX by intestinal bacteria in rats and volunteers and the active components in serum of rat after oral administration of DX 900mg/kg METHODS:To detect DX and its metabolites in stool,urine and serum with TLC and EST-MS RESULTS:In vitro,DX was decomposed easily by rat and human intestinal bacteria,and various metabolites were found With prolongation of metablism,metabolite with molecular weight of 415 3 was shown to be corresponding to diosgenin in rat serum and in urine of rats and volunteers CONCLUSION:The above-mentioned DX was decomposed easily by intestinal bacterias and diosgenin was absorbed into serum after oral administration of DX