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This study aimed to analyze the effect of Bletilla striata polysaccharide(BSP) on endogenous metabolites in serum of tumor-bearing mice treated with 5-fluorouracil(5-FU) by untargeted metabolomics techniques and explore the mechanism of BSP in alleviating the toxic and side effects induced by 5-FU. Male BALB/C mice were randomly divided into a normal group, a model group, a 5-FU group, and a 5-FU + BSP group, with eight mice in each group. Mouse colon cancer cells(CT26) were transplanted into the mice except for those in the normal group to construct the tumor-bearing mouse model by subcutaneous injection, and 5-FU chemotherapy and BSP treatment were carried out from the second day of modeling. The changes in body weight, diarrhea, and white blood cell count in the peripheral blood were recorded. The mice were sacrificed and sampled when the tumor weight of mice in the model group reached approximately 1 g. TUNEL staining was used to detect the cell apoptosis in the small intestine of each group. The proportions of hematopoietic stem cells and myeloid progenitor cells in bone marrow were measured by flow cytometry. Five serum samples were selected randomly from each group for untargeted metabolomics analysis. The results showed that BSP was not effective in inhibiting colon cancer in mice, but diarrhea, leukopenia, and weight loss caused by 5-FU chemotherapy were significantly improved after BSP intervention. In addition, apoptotic cells decreased in the small intestinal tissues and the percentages of hematopoietic stem cells and myeloid progenitor cells in bone marrow were significantly higher after BSP treatment. Metabolomics results showed that the toxic and side effects of 5-FU resulted in significant decrease in 29 metabolites and significant increase in 22 metabolites in mouse serum. Among them, 19 disordered metabolites showed a return to normal levels in the 5-FU+BSP group. The results of pathway enrichment indicated that metabolic pathways mainly involved pyrimidine metabolism, arachidonic acid metabolism, and steroid hormone biosynthesis. Therefore, BSP may ameliorate the toxic and side effects of 5-FU in the intestinal tract and bone marrow presumably by regulating nucleotide synthesis, inflammatory damage, and hormone production.
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Animals , Male , Mice , Colonic Neoplasms/drug therapy , Diarrhea , Fluorouracil/adverse effects , Hormones , Metabolomics , Mice, Inbred BALB C , Polysaccharides/pharmacologyABSTRACT
As the main active ingredient of the orchidaceous herb Bletilla striata, B. striata polysaccharide(BSP) has pharmacological activities such as promoting coagulation, anti-inflammation, anti-oxidation, promoting wound healing, anti-tumor, and immunomodulation, and is biodegradable and non-toxic. Additionally, it has the material properties of suspension thickening, film-forming adhesion, coating and solubilizing, targeting and slow releasing, effect-enhancing and toxicity-reducing, etc., playing the role of unification of medicines and excipients. Therefore, BSP has a wide application prospect in the fields of drug delivery system and trauma repair. This paper reviews the research progress of BSP application in new drug delivery systems and biomaterials based on the related li-terature in recent years, with the aim of providing reference for the further research and application of BSP.
Subject(s)
Biocompatible Materials , Drug Delivery Systems , Orchidaceae , Polysaccharides , Wound HealingABSTRACT
Objective: To prepare carboxymethyl Bletilla striata polysaccharide-chitosan@curcumin (CM-BSP) polyelectrolyte complex films, optimize their preparation technology, and evaluate its quality. Methods: CM-BSP was synthesized, then CM-BSP and CS formed water-insoluble complex by electrostatic bonding, the Cur-loaded polyelectrolyte complex films were prepared by a volatilization of solvent method. The formulation and preparation technology were optimized using an orthogonal design method and the morphology and structure were observed by scanning electron microscopy and fourier transform microscopic infrared spectroscopy. Results: The optimal prescription was of CM-BSP 117 mg, CS 233 mg, glycerol 25%, Cur 20 mg. The mean thickness of Cur-loaded polyelectrolyte complex films was (74.0 ± 2.0) μm, drug loading capacities was 95.41%, and in vitro release rate was 93.78%. Conclusion: The obtained polyelectrolyte complex films displayed an smooth exterior inspection, uniform distribution, good drug loading capacities and in vitro release rate.
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Objective: To optimize Bletilla striata polysaccharide (BSP)/polyvinylalcohol (PVA) wet spinning process by Box- Behnken response surface method and prepare the composite fiber and performe their structural characterization and performance evaluation. Methods: Taking the OD value of three types of mechanical property data (breaking force, breaking strength and elongation at break) of the fiber as the evaluation index, five factors (BSP mass fraction, PVA mass fraction, volumetric mix ratio of BSP/PVA, coagulation time and spinning speed) were investigated by single factor experiments. On the basis of the results of single factor experiments, three factors (BSP mass fraction, volumetric mix ratio of BSP/PVA, coagulation time) were investigated by response surface method to optimize BSP/PVA composite fiber wet spinning process. The morphology, structure, thermal property, and absorption property of the fibers were characterized and analyzed by SEM, IR, DSC, and water absorption test. Berberine hydrochloride (BH) was used as model drug to evaluate the drug loading property of the composite fiber and antibacterial activity of the drug loading fiber. Results: The optimal spinning process of composite fiber were as follows: BSP mass fraction was 7.5%, volume mixing ratio of BSP/PVA was 1:1 and coagulation time was 3 min. The composite fiber had a dense surface and formed a three-dimensional network structure inside, generated intermolecular forces, which enhanced the thermal and mechanical properties, and exhibited excellent water absorption capacity. The encapsulation efficiency of the composite fiber reached 70.2%. And the drug loading fiber formed obvious inhibition zone in the bacteriostatic zone test, which presented excellent antibacterial effect against E. coli and S. aureus. Conclusion: The optimized spinning process is feasible and low cost. The prepared composite fiber has better physical property and certain coating ability, and its application in the field of biomedical textiles is worth further study.
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OBJECTIVE： To study the mechanism of wound healing promotion of Panax notoginseng-Bletilla striata gum sponge on diabetic foot ulcer （DFU） model rats. METHODS： Healthy SD rats were selected and given high-lipid and high-glucose diet， intraperitoneal injection of streptozotocin once to establish diabetes model. Neodymium-iron-boron magnet was used to press the back of rats to make ulcer wound then established DFU model. Totally 60 DFU model rats were randomly divided into group A（blank group， i.e. normal saline gauze group）， group B（vaseline gauze group），group C （gelatin sponge group） and group D （P. notoginseng-B. striata gum sponge group）， with 15 rats in each group. The rats were given corresponding gauze/sponge to cover the wound for intervention treatment， changing dressing once every 1-2 days. On the 3rd and 7th day after intervention， the wound healing of rats in each group was observed with naked eyes， and the wound healing rate was calculated. The wound margin tissue was collected to obtain HE staining section， and histopathological observation was conducted under microscope. mRNA expression of β-catenin， GSK-3β and Rspo3 in wound tissue were determined by RT-PCR. RESULTS： On the 3rd and 7th day after intervention， compared with group A， B， C， healing rate of group D was increased significantly （P＜0.05）； inflammatory cell infiltration， collagen deposition， capillary and granulation tissue growth in wound tissue increased significantly. The mRNA expression levels of β-catenin and Rspo3 all increased， and those of GSK-3β all decreased； except for the difference of β-catenin at the 3rd day and GSK-3β at the 7th day after intervention between group D and group C were not significant， the difference of other indicators was statistically significant （P＜0.05）. CONCLUSIONS： P. notoginseng-B. striata gum sponge can effectively promote the wound healing in DFU model rats， the mechanism of which may be associated with up-regulating the expression of β-catenin and Rspo3 mRNA and down-regulating the expression of GSK-3β mRNA.
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OBJECTIVE： To investigate absorption kinetic characteristics of main active components as 4-（glucoseoxy）- glucoseoxybenzyl cinnamate （A1）， 2-isobutyl malic acid （A2）， 1，4-bis [4-（glucoxy） benzyl]-2-isobutyl malic acid ester （A3）， dihydrophenanthrenes 1 （A4） and 1，4-bis [4-（glucosoxy） benzyl]-2-isobutyl malic acid ester-2-（4-O-cinnamoyl-6-O-acetyl） glucoside （A5） from ethanol extract of Bletilla striata in the intestines of rats. METHODS： Using puerarin as internal standard， UPLC-MS/MS was used to determined the concentration of A1-A5 in intestinal circulation fluid. The determination was performed on Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile （containing 0.1％ formic acid）-water （containing 0.1％ formic acid） （gradient elution） at the flow rate of 0.35 mL/min. The column temperature was 45 ℃， and sample size was 3 μL. The positive ion and negative ion scanning were carried out in the multiple reaction monitoring mode by electrospray ion source. The ion pairs for quantitative analysis were m/z 593.2→431.1 （A1）， m/z 189.0→129.0 （A2）， m/z 725.3→457.2 （A3）， m/z 347.1→332.1 （A4）， m/z 1 059.3→793.1 （A5）， m/z 417.0→267.0 （internal standard）. In the in vivo intestinal circulation perfusion model， using accumulative absorption transfer rate （A） and absorption and transformation rate constant （Ka） as indexes， the effects of different doses of ethanol extract from B. striata （low-， medium-， high-dose were 166， 333，667 μg/mL，respectively）， bile， P-glycoprotein （P-gp） inhibitors （verapamil） and different intestinal segments on the absorption of above 5 components were investigated. RESULTS： The linear range of A1， A2， A3， A4 and A5 were 0.22-14.00， 0.34-21.75， 1.99-127.16， 0.15-9.75， 0.16-10.00 μg/mL（r＞0.99）. The limits of quantitation were 0.22， 0.34， 1.99， 0.15， 0.16 μg/mL， respectively. The lowest detection limits were 0.028， 0.085， 0.251， 0.035 and 0.010 μg/mL. RSDs of inter-day and intra-day were all lower than 10％. The recoveries ranged 83.60％-106.91％. Matrix effect did not affect the determination of the substance to be measured. A and Ka values of A1 in B. striata ethanol extract low-dose and medium-dose groups were significantly higher than high-dose group； A value of A3 in low-dose group was significantly higher than medium-dose and high-dose groups （P＜0.05 or P＜0.01）. A and Ka values of A1 and A3 in non-ligation group were significantly lower than control group， while A and Ka values of A4 were significantly higher than control group （P＜0.05 or P＜0.01）. A and Ka values of A1 and A3 in P-gp inhibitor group were significantly lower than control group （P＜0.05 or P＜0.01）. A values of A1 in jejunum group， ileum group and colon group， Ka value of A1 in colon group， A and Ka values of A2 in colon group， A value of A3 in ileum group， A and Ka values of A4 in ileum group and colon group， A values of A5 in jejunum group and ileum group as well as Ka value of A5 in jejunum group were all significantly lower than duodenum group. Ka values of A3 in jejunum group， ileum group and colon group were significantly higher than duodenum group （P＜0.05 or P＜0.01）. CONCLUSIONS： Established UPLC-MS/MS method is specific， sensitive and simple， and it can be used for quantitative analysis and pharmacokinetic study of A1-A5. The 5 active components in B. striata ethanol extract are absorbed by the whole intestine， and the intestinal segments are different. A1 and A3 are absorbed more in intestinal tract and may be saturated. Bile can inhibit intestinal absorption of A1 and A2， but promoted intestinal absorption of A4. A1-A5 may not be the substrate of P-gp.
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OBJECTIVE： To study in vitro metabolism pathway of effective component of Bletilla striata as Militarine in liver microsomes and kinetics characteristics of enzyme-catalyzed reactions. METHODS： The in vitro incubation system of rat and human liver microsomes was established， and incubation reaction of Militarine was performed. UPLC-QTOF-MS was used to identify the structure of its metabolites in combination with UNIFI database and references. Using puerarin as internal standard， UPLC-Triple Quad-MS was used to quantitatively analyze metabolic transformation of Militarine in rat liver microsomes. The kinetic parameters （vmax， km， CLint） of Militarine enzyme-catalyzed reactions with/without reducing coenzyme Ⅱ （NADPH） were calculated by fitting the curves with GraphPad Prism 5.0 software. RESULTS： After incubation in rat and human liver microsomes， Militarine produced a chemical formula C21H29O11， which was presumed to be a metabolite of Militarine ester bond hydrolysis. The kinetic study of enzyme-catalyzed reactions showed that vmax of Militarine enzyme-catalyzed reactions with/without NADPH were 1.955， 2.129 nmol/（h·mg）； km were 8.601， 9.854 nmol/mL； CLint were 0.227 3， 0.216 1 mL/（h·mg）； there was no significant difference between with NADPH and without NADPH. CONCLUSIONS： The main metabolic pathway of Militarine in liver microsomes is the hydrolysis of C1 and C4 ester bonds. Its metabolism does not depend on the pathway of cytochrome P450 enzymes initiated by NADPH.
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To investigate the active fraction from Bletilla striata in Caco-2 cell monolayer,so as to explore its absorption mechanism of oral administration preliminarily.Active fraction from B.striata in Caco-2 cell monolayer was analyzed by UPLC-Q-TOF and detected by UPLC-MS/MS,and the effects of different concentrations,pH and P-glycoprotein inhibitors on Caco-2 cells Monolayer were investigated.Six compounds were isolated from the active fraction of B.striata in Caco-2 cell monolayer by UPLC-Q-TOF,and identified as B6,B12,B14,B17,B19 and B23,with concentration dependence.Within the 0-180 min,the uptake of B12 and B14 had a time dependence,while B6,B17,B19 and B23 tended to saturate after 60 min.All of the components had a good absorption in an acidic environment.B6 had a good absorption at pH 6.0,while the other components B12,B14,B17,B19 and B23 had a good absorption at pH4.0.The absorption of the 6 main components of B.striata were not be affected by P-glycoprotein inhibitors(verapamil/cyclosporin A).Compared with the control group,there was no difference in the absorption of B6 and B12,and the absorption of B14,B17,B19 and B23 increased,but with no significant difference.The absorption characteristic of B.striata extract across the Caco-2 cell monolayer is probably passive diffusion,and the absorption process was not affected by P-glycoprotein.
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Humans , Biological Transport , Caco-2 Cells , Chromatography, Liquid , Intestinal Absorption , Orchidaceae , Chemistry , Plant Extracts , Pharmacology , Tandem Mass SpectrometryABSTRACT
OBJECTIVES@#To evaluate the characteristics of Bletilla striata microspheres (BSMs) and its effects as an embolic agent in a rabbit model.@*METHODS@#BSMs were prepared with an emulsification-cool condensation-chemical cross-linking method. The characteristics of BSMs in vitro were observed. Embolization experiments were performed in renal artery of rabbit and in a rabbit liver VX2 carcinoma model. Seventy-two New Zealand rabbits were divided into 2 groups, and the right renal artery was embolized with BSMs (200 μm in diameter) in the experimental group and with polyvinyl alcohol (PVA) of the same size in the control group. The pathological findings were examined with hematoxylin-eosin and Masson stainings. Liver and renal functions were tested before and after embolization. VX2 tumor was transplanted in 15 New Zealand rabbits, which were randomly divided into 3 groups (n=5). Group A were treated with saline, group B with a mixture of doxorubicin and lipiodol, and group C with hepatic arterial infusion of BSMs (200 μm in diameter). Tumor growth rate was evaluated by magnetic resonance imaging scan. Apoptosis-related factors (bax, bcl-2) and tumor vascular endothelial cell growth factor (VEGF) were evaluated through immunohistochemical staining.@*RESULTS@#The characteristics of BSMs in vitro were in full compliance with the requirements for use in interventional procedures. In the renal artery embolization experiment, after BSMs intervention, it was more difficult to form collateral circulation than that with PVAs, and the kidney manifested atrophy and calcification. There were no significant difference of liver and renal functions in rabbits between groups. In the liver VX2 carcinoma embolization experiment, compared with group A, the growth rate of VX2 liver tumor and Bcl-2 levels was reduced, while apoptosis index, Bax, and VEGF were increased in group B (P0.05).@*CONCLUSIONS@#The characteristics of BSMs in vitro and in vivo meet the requirements for its use as an embolic agent in interventional approaches.
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In order to understand the difference of contents of coelonin,batatasin Ⅲ and 3'-O-methylbatatasin Ⅲ in 60 different sources of Bletilla striata planted under the same conditions. UPLC method was used and the analysis was performed on a Waters ACQUITY UPLC BEH C18 column( 2. 1 mm×100 mm,1. 7 μm),eluted with acetonitril-0. 1% formic acid solution by gradient. The flow rate was 0. 208 m L·min-1,the detection wavelength was 270 nm,the column temperature was 35 ℃ and the injection volume was 4μL. Under the above chromatographic conditions,the three components can be separated well with good linearity in the range of 0. 156-5. 000 mg·L-1. The average contents of coelonin,batatasin Ⅲ and 3'-O-methylbatatasin Ⅲ were( 0. 116 ± 0. 071) %,( 0. 386 ±0. 185) % and( 0. 086±0. 034) %,respectively. After planting for two years under the same conditions,there was no significant difference in chemical composition among different sources and varieties,but the contents of the three components had some regional differences,which indicated that the western region was higher than the eastern region,while the contents of coelonin and batatasin Ⅲ in B.sinensis were slightly higher than those in B. striata. The chromatographic method above is simple,stable and reproducible,and can be used for quantitative analysis of three components. The content analysis of different sources of B. striata can provide reference for future B. striata breeding and quality control.
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Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Chemistry , Orchidaceae , Chemistry , Quality ControlABSTRACT
Objective: Phenylalanine ammonia-lyase gene (PAL) is one of the key enzymes associated with stress resistance in secondary metabolism pathway of plants. Exploring its sequence information and expression profiling information in stress response could comprehensively peep at the protein structure, functions and signal network of plant stress resistance. Methods: The full cDNA length of PAL from Bletilla striata was cloned by RT-PCR and RACE approaches. Physicochemical properties and conserved domain of BsPAL protein were determined by a series of bioinformatics tools as Protparam, SOPMA, SWISS-MODEL, etc. Multiple alignment and phylogenetic tree were achieved by DNAMAN and MEGA Software, respectively. The qPCR was employed to examine the expression profiles of BsPAL under exogenous hormone stress. Results: The full cDNA of BsPAL was 2 708 bp, encoding a 797 amino-acid protein with a molecular weight of 86 216.94 and an isoelectric point (pI) of 6.24. The BsPAL protein included the typical structural domain and active site of PALs in other plants, and without transmembrane region, which was more homologous with PALs of Dendrobium officinale and Phalaenopsis aphrodita. The qPCR Results: revealed the expression level of BsPAL in roots was much higher than that in leaves and stems. Under MeJA treatment, the expression trend of BsPAL was first gradually ascending and then descending, while SA treatment had the reverse effect. Conclusion: The BsPAL’s sequence characterizing, expression profiling and responding patterns against SA and MEJA provided a research basis for elucidating the metabolic pathways of phenylpropanoid and hormone signaling research in B. striata.
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Objective: The genetic diversity of wild Bletilla striata from different sources was analyzed in order to provide the reference value for breeding, protection and development of B. striata germplasm in the future. Methods: Genetic diversity of 50 wild B. striata samples was identified by SRAP molecular marker technique, and the genetic relationship was analyzed by UPGMA method. Results: A total of 117 bands were amplified from 50 samples by using 15 SRAP primer combinations, with an average of 7.8 bands per primer. Among them, 106 bands were polymorphic, with a polymorphism rate of 90.60%. The clustering results showed that the genetic similarity coefficient of 50 B. striata samples was 0.59-0.87, which could be divided into six categories based on the genetic coefficient of 0.66. The genetic distance of two pieces of B.striata from No.2 Lijiang, Yunnan and No.2 Jingshan County, Hubei was not directly related to geographical distance. Conclusion: There are abundant genetic diversity among wild B. striata resources, and there is no direct relationship between geographical distance and genetic distance. SRAP technology can provide theoretical basis for the protection of B. striata resources and variety breeding, which provides reference for the subsequent development and utilization of B. striata.
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Objective: Bletilla striata polysaccharide (BSP-1) extracted from the stem tubers of B. striata was purified to study the structural properties and antitumor activity. Methods: The crude polysaccharide was fractionated by DEAE-cellulose column chromatography, and three fractions were obtained including BP-1, BP-2, and BP-3, respectively, BP-1 was further purified by SephadexG-200 column chromatography, and a sub-fraction named BSP-1 was obtained. Subsequently, HPGPC, IR spectroscopy, gas chromatography (GC), GC coupled to mass spectrometry (GC-MS) and methylation analysis, and 1H- and 13C-NMR were employed to characterize the structural properties of the polysaccharide fractions. Moreover, the anticancer activity was investigated in vivo. Results: The results showed that the molecular weight of BSP-1 were 4.72 × 105. BSP-1 was consisted of mannose (Man) and glucose (Glc) residues. The backbone of BSP-1 was composed of β-1,4-linked Man and Glc residues at the end with a molar ratio of 8:1. BSP-1 could inhibit the tumor proliferation of HepG2-bearing mice. The tumor weight of mice was significantly inhibited by BSP-1 with inhibitory rate of 66.42%. Conclusion: Structural characterization of BSP-1 provides the significant reference for the further development and elucidation of polysaccharides from B. striata as new drugs.
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Objective: Because of the adhesion of Bletilla striata polysaccharide (BSP), it was mixed with sodium alginate (SA) as a composite carrier to prepare mucoadhesive PNS-BSP composite microspheres. Panax notoginseng saponins (PNS) dispersion with sustained release property was used as an encapsulating drug. Methods: The composite microsphere was prepared by ion cross-linking method. The formulation process was investigated and optimized by single factor test and orthogonal design. The microspheres were evaluated by scanning electron microscope (SEM), particle size distribution, DSC, swelling properties, in vitro mucoadhesive properties, and in vitro release characteristics. Results: PNS-BSP composite microspheres had good roundness, rough surface and wrinkles. The microspheres showed a narrow size distribution. PNS was uniformly dispersed in microspheres in an amorphous state. The microspheres prepared by the best prescription process were stable in process and reproducible. Compared with the microspheres prepared by directly adding PNS, the drug loading, encapsulation efficiency and yield of PNS dispersion microspheres were increased significantly, which were 10.34%, 51.25%, and 82.21%, respectively. The drug loading, encapsulation efficiency, and yield were 4.04%, 12.16%, and 61.35% of PNS microspheres. The addition of BSP increased the swelling properties of the SA microspheres, and significantly increased the retention rate in the stomach of rats. The release of ginsenoside Rg1 in PNS-BSP microspheres was released slower compared to PNS. Conclusion: The bioadhesion of microspheres was increased by the addition of BSP. The drug loading, encapsulation efficiency, and yield of the microspheres were increased by the preparation of PNS as a dispersion, and the microspheres also had a certain sustained-release effect.
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Objective: To study the glycoside chemical constituents of Bletilla striata grown in Guizhou Province. Methods: The glycoside chemical constituents were separated and purified by silica gel column chromatography, MCI, gel column chromatography, medium pressure preparation, highperformance liquid chromatography and other modern separation methods. The structures were identified based on the spectral data. Results: Thirteen glycosides were isolated from the 95% ethanol extracts: 1-methyl-3-phenylpropyl-β-D-glucopyranoside (1), 1-(4-β-D-glucopyranosyl oxybenzyl) 4-ethyl (2R)-2-isobutylmalate (2), 3’,5- dimethoxy-bibenzyl-3-O-β-D-glucopyranoside (3), syringaresinol mono-β-D-glucoside (4), 4-O-(6’-O-glucosyl-p-coumaroyl)-4- hydroxybenzyl alcohol (5), (4-hydroxyphenyl) methyl-β-D-glucopyranoside (6), 4-methylphenyl-β-D-glucopyranoside (7), benzyl-β-D- glucopyranoside (8), phenyl-β-D-glucopyranoside (9), 4-[(acetyloxy) methyl] phenyl-β-D-glucopyranoside (10), batatasin III-3-O- glucoside (11), gastrodin (12) and 4-(4-β-D-glucopyranosyl-oxybenzyl)-(2R)-2-isobutylmalate (13). Conclusion: Compound 1 is a new natural product. Compounds 3-6 are isolated from this plant for the first time; Compounds 2, 7-13 are obtained from this genus for the first time.
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Rhizoma Bletillae is in high demand as a traditional Chinese medicine, and the natural resources have been severely damaged due to excessive exploitation. Because Bletilla striata seeds are small and have no endosperm, the seed germination rate is low in natural conditions. Traditional division propagation could not meet the demands of large-scale cultivation. Tissue culture can provide many seedlings in a short time and is more effective and convenient than other methods. Most studies on tissue culture of B. striata selected seeds as explants. This review summarized the processes of the aseptic seed germination pathway. It included such stages as seed germination, proliferation of clusters of buds, induction of rooting and transplanting of seedlings. Influential factors as well as optimum combination and concentration of the plant growth regulators of each stage were also summarized. The further research on tissue culture of B. striata was also prospected.
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Objective To prepare palmatine-loaded flexible nano-liposomes (PFL) films with Bletilla striata polysaccharide (BSP) as membrane material, and evaluate its pharmacy related performance in order to lay the foundation for further application. Methods The PFL was prepared by injection method and the films of PFL based on Bletilla striata polysaccharide (PFL-BPF) was prepared by homogenate coating method. The PFL-BPF was characterized and evaluated by electron microscopy, differential scanning calorimeter (DSC), and in vitro transmucosal membrane experiment. Results The PFL and BSP had good compatibility and easy to film with BSP as membrane material. The appearance of PFL-BPF obtained was smooth, non-bubble, flexible, and suitable stiffness; PFL-BPF had good biological adhesion. The time of scouring the film agent from mucous membrane with normal saline was (130 ± 7) min. At 0.5 h, the dose of PFL-BPF promoting palmatine (PA) infiltration to mucosa was 32.41 μg/g. It was 3.17 times higher than those of PA solution based on BSP (PL-BPF) and 1.9 times for PA common liposomes based on BSP (BLP + PA-BPF) (t-test, P < 0.05); At 2.5 h, it was 2.67 times and 1.89 times higher than those of PL-BPF and BLP + PA-BPF, respectively. It showed that PFL-BPF could significantly promote the water-soluble drug PA through mucosa membrane and release it slowly. The results of DSC showed that the possible mechanism for promoting the absorption of PA through mucosa membrane was that the flexible liposomes disturbed the mucosal epithelial cells and carried the drug into the mucosal tissue. Conclusion The PFL-BPF had the advantages of good film-forming property, lasting adhesive attraction, strong scour resistance, simple and feasible preparation process, and could promote drug permeation into mucosa obviously. Therefore, the flexible nano-liposomes film is a good drug carrier for the transmucosal drug delivery applications and has a wide application prospect.
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Objective To prepare reactive oxygen species (ROS)-responsive Bletilla striata polysaccharide (Oxi-BSP) micelles, optimize their preparation technnology, and evaluate its in vitro characterizations. Methods Oxi-BSP was synthesized, then curcumin (Cur)-loaded micelles were prepared by a dialysis method. The formulation and preparation technology was optimized using an orthogonal design method. The morphology was observed by transmission electron microscope, while the particle size, particle distribution, and zeta potential were determined by laser particle size analyzer. Hydrogen peroxide was used to simulate the ROS environment and then the change of micellar morphology was observed. Results The Cur-loaded micelles were spherical with homogeneous distribution, the mean size was (225.33 ± 2.97) nm, the zeta potential was (-16.80 ± 0.37) mV, the encapsulation efficiencies was (85.75 ± 0.87)%, and drug loading capacities was (20.21 ± 0.44)%. And the micelles were responsive to ROS stimuli. Conclusion The obtained micelles display an uniform particle size distribution with moderate drug encapsulation efficiencies and drug loading capacities.
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Microspheres (MS) are an excellent transarterial chemoembolization carrier for cancer treatment. Then the Bletilla striata polysaccharide (BSP) that was isolated from the rattan of Bletilla striata was used as skeleton material, and the matrine (ME) loaded Bletilla striata polysaccharide microspheres (ME-BSPMS) were prepared by emulsify-chemical crosslinking method. ME-BSPMS was characterized for appearance shape, particle size, drug loading, swelling ratio, suspension property, drug entrapment condition and in vitro release characteristics. The results showed that the ME-BSPMS appeared as round spherical and smooth shape by SEM, with an average size of (85 ±7) μm. ME-BSPMS with a good suspension in physiological saline and the swelling ratio could reach upwards of (53 ±4.2)% in 20 minutes, also with a large amount of drug loading of (30.12 ±3.25)%. The results of DSC scanning indicate that good compatibility exists between the ME and BSP, and the ME could be embedded fully in the matrix of the ME-BSPMS. The accumulation drug release from ME-BSPMS was (25.38 ±1.57)% at 12 h, this suggests that the ME-BSPMS has a good sustained release effect. These results indicate that the ME-BSPMS may be a promising transarterial chemoembolization carrier for cancer treatment.
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OBJECTIVE:To set up a method for determination of molecular weight and content of polysaccharide in Bletilla striata. METHODS:HPSEC-ELSD method was adopted. The determination was performed on TSK-GEL G4000 PWXL column with mobile phase consisted of pure water at the flow rate of 0.6 mL/min.The column temperature was 30 ℃,and sample size was 20 μL. Evaporative light scattering detector was used.The molecular weight and content of polysaccharide in 3 batches of B. striata were established. RESULTS:The linear range of weight average molecular weight of B. striata polysaccharide were 24.17-178.00 kD(R2=0.985 5). The linear range of BT07 content determination were 0.508 0-5.080 mg/mL(R2=0.998 4). The limits of detection and quantitation were 0.116 5 and 0.274 0 mg/mL. RSDs of precision,stability and reproducibility tests were all lower than 2%(n=6 or n=7).Average recoveries rate were 99.16%-100.20%(RSD=0.39%-0.64%,n=9).Average retention time of 3 batches of samples was 14.28 min(RSD=4.35%,n=3),and weight average molecular weight was 23.54 kD(RSD=3.78%, n=3). Polydispersity coefficient D,MW/Mn)ranged 1.463-1.578. Average content of sample was 93.4%(RSD=4.22%,n=3). CONCLUSIONS:HPSEC-ELSD method is simple,rapid,accurate and reliable,which can be used for simultaneous determination of B.striata polysaccharide.