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Aim To evaluate the hypolipidemic effect of the total phenylpropanoid glycosides extracted from Ligustrum robustum (Roxb.) Blume (LRTPG) on hyperlipidemic golden hamsters and explore its regulatory effect on intestinal flora. Methods Sixty hamsters were randomly divided into a control group, a model group, a positive drug group, LRTPG-L group, LRTPG-M group, and LRTPG-H group. After the successful induction of the model by high-fat diet, the animals were continuously administered for four weeks, and their blood lipids and liver lipids were detected. The formed feces from the colorectal region of the hamsters in the control group, model group and LRTPG-H group were collected for 16S rDNA sequencing. Results LRTPG reduced serum TG, TC, LDL-C and liver TG, TC concentrations significantly in hyperlipidemic hamsters. The results of the intestinal microbiota sequencing showed that compared to the control group, LRTPG significantly decreased the relative abundance of the phylum Firmicutes and increased the relative abundance of the phylum Bacteroidetes and Verrucomicrobia (P < 0.01) at the phylum level. At the family level, LRTPG significantly increased the relative abundance of Christensenellaceae, Peptococcaceae, and Verrucomicrobiaceae (P < 0.05 or P < 0.01). At the genus level, LRTPG significantly increased the relative abundance of Oscillospira, Oscillibacter, Flavonifractor and Akkermansiaceae (P < 0.05 or P < 0.01). These changes in the flora were beneficial to the hypolipidemic effect of LRTPG. Conclusion LRTPG may exert its hypolipidemic effect by improving the intestinal flora disorder caused by a high-fat diet in golden hamsters.
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Ligustrum lucidum is a woody perennial plant of genus Ligustrum in family Oleaceae. Its dried fruit has high medicinal value. In this study, the authors evaluated the variability and species identification efficiency of three specific DAN barcodes(rbcL-accD, ycf1a, ycf1b) and four general DAN barcodes(matK, rbcL, trnH-psbA, ITS2) for a rapid and accurate molecular identification of Ligustrum species. The results revealed that matK, rbcL, trnH-psbA, ITS2 and ycf1a were inefficient for identifying the Ligustrum species, and a large number of insertions and deletions were observed in rbcL-accD sequence, which was thus unsuitable for development as specific barcode. The ycf1b-2 barcode had DNA barcoding gap and high success rate of PCR amplification and DNA sequencing, which was the most suitable DNA barcode for L. lucidum identification and achieved an accurate result. In addition, to optimize the DNA extraction experiment, the authors extracted and analyzed the DNA of the exocarp, mesocarp, endocarp and seed of L. lucidum fruit. It was found that seed was the most effective part for DNA extraction, where DNAs of high concentration and quality were obtained, meeting the needs of species identification. In this study, the experimental method for DNA extraction of L. lucidum was optimized, and the seed was determined as the optimal part for DNA extraction and ycf1b-2 was the specific DNA barcode for L. lucidum identification. This study laid a foundation for the market regulation of L. lucidum.
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Ligustrum/genetics , Seeds , Fruit , Polymerase Chain Reaction , Research DesignABSTRACT
Ligustri Lucidi Fructus, the sun-dried mature fruit of Ligustrum lucidum, is cool, plain, sweet, and bitter, which can be used as both food and medicine, with the effects of improving vision, blacking hair, and tonifying liver and kidney. It takes effect slowly. However, little is known about the genetic information of the medicinal plant and it is still a challenge to distinguish Ligustrum species. In this study, the complete chloroplast genome of L. lucidum was obtained by genome skimming and then compared with that of five other Ligustrum species, which had been reported. This study aims to evaluate the interspecific variation of chloroplast genome within the genus and develop molecular markers for species identification of the genus. The result showed that the chloroplast genome of L. lucidum was 162 162 bp with a circular quadripartite structure of two single-copy regions separated by a pair of inverted repeats. The Ligustrum chloroplast genomes were conserved with small interspecific difference. Comparative analysis of six Ligustrum chloroplast genomes revealed three variable regions(rbcL-accD, ycf1a, and ycf1b), and ycf1a and ycf1b can be used as the species-specific DNA barcode for Ligustrum. Phylogeny analysis provided the best resolution of Ligustrum and supported that L. lucidum was sister to L. gracile. This study clarified the genetic diversity of L. lucidum from provenance, which can serve as a reference for further analysis of pharmacological differences and breeding of excellent varieties with stable drug effects.
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Fruit , Genome, Chloroplast , Ligustrum/genetics , Phylogeny , Plant BreedingABSTRACT
Aim To explore the anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription based on network pharmacology. Methods The active components and targets of Shuanghuang Qizhen Keliu Prescription were searched and selected through the TCM-SP database. Related targets of tumor were screened and retrieved by OMIM. The key anti-tumor targets of Shuanghuang Qizhen Keliu Prescription were obtained by using Venn diagram analysis. The key target PPI network was constructed and the key targets were obtained by Cytoscape. Metascape platform was used for GO and KEGG enrichment analysis of key targets. Results There were many active components such as quercetin, kaempferol, and beta-sitosterol in Shuanghuang Qizhen Keliu Prescription, and their targets might be TP53, AKT1, JUN, MAPK1, TNF, RELA, IL-6 and so on. The anti-tumour mechanism might be proliferation inhibition, migration inhibition and apoptosis promotion. Conclusions This study preliminarily discusses the potential anti-tumour mechanism of Shuanghuang Qizhen Keliu Prescription, providing ideas and basis for subsequent experimental research.
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OBJECTIVE：To i mprove the transfer rate and purity of oleanolic acid and ursolic acid in total triterpenoids from Ligustrum lucidum ，so as to optimize the purification technology. METHODS ：Oleanolic acid and ursolic acid were used as representative components of total triterpenoids ，and their contents were determined by HPLC. The determination was performed on Thermo BDS Hypersil C 18 column with mobile phase consisted of methanol- 0.02％ ammonium acetate solution （80∶20，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm，and column temperature was 30 ℃. The sample size was 20 μ L. In single factor tests，using transfer rate of oleanolic acid and ursolic acid as index ，the effects of water precipitation temperature and time ，the amount of redissolved ethanol on the purification technology was investigated ；using transfer rate and purity of two components as indexes ，the effects of the amount of activated carbon and volume fraction of crystallization ethanol were investigated. Based on it ，using the amount of redissolved ethanol and activated carbon ，volume fraction of crystallization ethanol as factors ，Box-Behnken response surface methodology was used to optimize the purification technology ，and validation tests were performed. RESULTS ：The optimal purification technology was adding 4-fold（mL/g，the same below ）water in L. lucidum concentrated solution ，placing for 2 hours at 0 ℃（water precipitation ）；adding 1-fold ethanol to dissolve （redissolution）； adding 4％ activated carbon （edulcoration）；finally adding water to adjust the volume fraction of ethanol to 80％，placing at 4 ℃ for 12 hours（crystallization），centrifuging and drying. The results of 3 times of validation tests showed that the transfer rates of oleanolic acid and ursolic acid in total triterpenoids prepared by optimized technology were 61.11％ and 65.78％，the purities of them were 53.44％ and 19.79％，and RSDs were both lower than 3％. CONCLUSIONS ：The optimized purification technology has high extraction efficiency and simple operation ，which can be used for industrial production of purification of total triterpenoids from L. lucidum and the development of corresponding preparations.
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OBJECTIVE@#To examine the effect of the aqueous extract of Ligustrum robustum on tumor growth in vitro and in vivo and explore the possible molecular mechanisms.@*METHODS@#In in vitro study, cell viabilities of human cervical carcinoma cells (HeLa), human breast cancer cells (MCF-7), human prostate cancer cells (PC-3), human hepatoma cells (7721) and human colon carcinoma cells (SW480) were evaluated with cell counting kit-8. For L. robustum-treated Hela cells, early or late apoptosis were evaluated by annexin V/PI staining. Mitochondrial membrane potential was measured by staining cells with JC-1. Apoptosis was monitored by nuclear morphology based on chromatin condensation and fragmentation by 4',6-diamidino-2-phenylinole (DAPI) staining. Caspase-3 and -8 activity levels were measured by a colorimetric assay. In vivo, to evaluate the possible mechanism of L. robustum-mediated antitumor effect, nude mouse xenograft study was also conducted.@*RESULTS@#In in vitro study, L. robustum was found to be toxic to HeLa, MCF-7, PC-3, 7721, SW480, with an half maximal inhibitory concentration value of 2-5 mg/mL (P<0.05). Moreover, externalization of phosphatidylserine, loss of mitochondrial membrane potential, DNA fragmentation and activation of caspase-3 and -8 were detected in L. robustum-treated Hela cells. Using a nude mouse model bearing Hela xenografts, we found that L. robustum reduced tumor volume and tumor weight (P<0.05), but had no effect on body weight and histological damage of important organs. Intraperitoneal injection of L. robustum caused a significant reduction in serum aspartate transaminase and alanine transaminase levels (P<0.05). Furthermore, cleaved caspase-3-positive and terminal nucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells were observed in L. robustum-treated tumor tissues.@*CONCLUSIONS@#L. robustum inhibits tumor cell growth both in vitro and in vivo by inducing apoptosis in a caspase-dependent way without apparent hepatic toxicity and histological damage, which may offer partial scientific support for the ethnopharmacological claims of L. robustum as a herbal tea for its antitumor activity.
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Aim To explore the hypolipidemic mechanism of the total phenylpropanoid glycoside from Ligustrum robustum (Roxb. ) Blume (LRTPG) on hyperlipidemic hamsters using label-free quantitative proteomic technique. Methods The total protein was extracted from livers of model group and the group treated with LRTPG for label-free quantitative proteomics research. Results The proteomic data showed that a total of 2231 proteins were identified. And 549 proteins were found to be differentially expressed between model group and group treated with LRTPG. Among the 549 proteins, 93 proteins were up-regulated and 59 proteins were down-regulated, and 397 proteins had quantitative values only in model group or drug-administered group. Further, gene ontology (GO) analysis indicated that those differentially expressed proteins were primarily involved in an array of biological processes including metabolism, transport, oxidation-reduction, phosphorylation, signal transduction and lipid metabolism. KEGG pathway analysis revealed that these proteins were involved in several signal pathways including oxidative phosphorylation, non-alcoholic fatty liver dis-ease, PI3K-Akt, cAMP, and cGMP-PKG pathway. And some of these proteins were much related to the lipid metabolism, such as CD36, PK, HSS, GCK, ApoA I, Acly and FABP5. Conclusion The hypolipidemic effect of LRTPG may be related to CD36, PK, HSS, GCK, ApoA I, Acly and FABP5.
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Objective: To investigate the relationship between the anti-osteoporotic effect of Ligustrum lucidum and the growth hormone (GH)/insulin-like growth factor 1 (IGF-1) signaling pathways. Methods: L. lucidum aqueous extract was orally administrated to ovariectomized (OVX) rats for 14 weeks. Then the femurs were removed and stained with hematoxylin & eosin (HE) and Safranin O/Fast Green staining, respectively, to evaluate the change of bone microstructure. The histomorphological parameters and bone mineral density (BMD) of the femurs were measured by micro-CT. Furthermore, rat serum GH level was determined by ELISA assay, and IGF-1 protein expression in liver and bone was determined by Western blotting and immunohistochemical staining. Results: L. lucidum prevented the disorganized femoral trabeculae and inhibited the decrease in BMD and glycosaminoglycan content in OVX rats. In addition, L. lucidum significantly inhibited the decrease of serum GH levels and improved IGF-1 protein expression of liver and bone in OVX rats. Conclusion: L. lucidum may prevent against osteoporosis through inhibition of bone loss and improvement of bone microstructure via regulating GH/IGF-1 signaling pathway.
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Objective: To study the relative molecular mass and composition of Ligustrum lucidum polysaccharide purified by dialysis, and provide the theories for the relationship between the biological activity and the internal composition. Methods: In this paper, L. lucidum as an object was studied. Polysaccharide was isolated by water extraction and ethyl alcohol precipitation and purified by Sevage method, hydrogen peroxide and dialysis. After the acidolysis of trifluoroacetic acid, the molecular weight was measured by GPC, and the composition of polysaccharide was analyzed by HPLC-RID. Results: The Mw of polysaccharide was 10 721, and the Mn of polysaccharide was 10 673. L. lucidum polysaccharide consisted of glucose, rhamnose, and arabinose, the molar ratio of these monosaccharide was 9.148.105.18. Conclusion: The purified polysaccharide composition is more homogeneous, and the monosaccharides of polysaccharides was easily analyzed by HPLC-RID without column derivatization.
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Background: Thorough pharmacological experiments on various plants used in traditional medicines are in progress in order to establish their effectiveness and safety. But modern drugs or conventional medicines are often viewed as impersonal, emphasizing crisis intervention. Keeping in view the above idea, the present study is undertaken on the plant Ligustrum robustum to explore its antipyretic property in albino rats.Methods: Healthy young albino rats weighing between 100-250gm were obtained for the study. The animals were divided into five groups with six animals in each group. Pyrexia was induced by subcutaneous injection of dried yeast in 2% gum acacia in normal saline at a dose of 20ml/kg body weight below the nape of the neck in albino rats. The antipyretic activity of the aqueous extract of Ligustrum robustum was tested by yeast induced method. The data were subjected to ANOVA followed by Dunnett’s ‘t’ test for statistical significance between different groups.Results: Ligustrum robustum in doses of 100mg/kg, 200mg/kg and 400mg/kg significantly reduced the temperature (p<0.05 to 0.01) in the 3rd and 4th hour after drug administration.Conclusions: Present study concludes that the aqueous extract of the leaves of Ligustrum robustum showed significant antipyretic property in the standard experimental animal models.
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According to the previous results from transcriptome analysis of Ligustrum quihoui, a glycosyltransferase gene(xynzUGT) was cloned by rapid amplification of cDNA ends(RACE). The full length cDNA of xynzUGT was 1 598 bp, consisting of 66 bp 5'-UTR, 1 440 bp ORF and 92 bp 3'-UTR. The ORF encoded a 480 amino-acid protein(xynzUGT) with a molecular weight of 54 826.67 Da and isoelectric point of 5.82. The structure of enzyme was analyzed by using bioinformatics method, the results showed that the primary structure contained a highly conserved PSPG box of glycosyltransferase, the secondary structure included α helix(38%), sheet(12.1%) and random coil(49.9%), and tertiary structure was constructed by peptide chain folding to form two face-to-face domains(often referred to as a Rossmann domains), between which a substrate binding pocket is sandwiched. The phylogenetic tree analysis indicated that xynzUGT might catalyze glycosylation of phenylpropanoids, such as tyrosol. Further simulation experiment of molecular docking between enzyme and tyrosol showed that Gly138 and Ser285 located in the binding pocket interacted with tyrosol by hydrogen bonding. SDS-PAGE analysis exhibited that the prokaryotic expression system successfully expressed recombinant xynzUGT with molecular weight of 58 370.57 Da, but it exists in the form of non-soluble inclusion bodies. Using the molecular chaperone and enzyme co-expression method, the soluble expression was promoted to some extent. The above works laid the foundation for further studying on enzymatic reaction and clarifying the functional mechanism of enzyme.
Subject(s)
Cloning, Molecular , DNA, Complementary , Glycosyltransferases , Genetics , Ligustrum , Genetics , Molecular Docking Simulation , Phylogeny , Plant Proteins , Genetics , Protein Structure, Secondary , Protein Structure, TertiaryABSTRACT
OBJECTIVE To explore the hypolipidemic mechanisms of the total phenylpropanoid glycosides from Ligustrum robustum (Roxb.)Blume (LRTPG)in hamsters using proteomics technique. METHODS The hamsters were fed with a high fat diet to induce hyperlipidemia.Then LRTPG of high (1.2 g·kg-1),medium(0.6 g·kg-1)and low(0.3 g·kg-1)doses were administrated daily for 4 weeks.Then the concentrations of plasma and hepatic lipids were determined using enzymic methods.The total protein was extracted from livers of the model group and the group treated with the high dose of LRTPG for label-free quantitative proteomics. RESULTS LRTPG significantly reduced the concentrations of plasma and hepatic lipids in hamsters fed a high fat diet. The proteomics data showed that a total of 2231 proteins were identified,and 549 proteins were found to be differentially expressed between the model group and the group treated with LRTPG.Among the 549 proteins,93 proteins were up-regulated and 59 proteins were down-regulated, and 397 proteins were absent or not. And some of these proteins were much related to the lipid metabolism. Further, gene ontology (GO) analysis indicated metabolic process, transport, oxidation-reduction process, phosphorylation, signal transduction, lipid metabolic process were the main biological processes that those differentially expressed proteins participated. KEGG pathway analysis showed that those proteins were involved in several metabolic pathways including oxidative phosphorylation,non-alcoholic fatty liver disease(NAFLD),PI3K-Akt signaling pathway, cAMP signaling pathway, cGMP-PKG signaling pathway. CONCLUSION The proteomics study could provide valuable clues to help us to understand the hypolipidemic mechanisms of LRTPG much better.
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Medicinal plants are potential sources of anticancer agents screening. A large number of phytochemicals, including triterpenoids, have been reported to have significant cytotoxic effects on cancer cells. From the fruits of Ligustrum japonicum Thunb., thirteen triterpenoids (1 – 13) were isolated and evaluated for their cytotoxic activity against Hela and HL-60 cells. As results, 8 (oleanolic acid) showed significant effects on Hela with IC50 values of 5.5 µM, and moderate effects on HL-60 cells with IC₅₀ values of 55.9 µM. Meanwhile, 10 (oleanderic acid) and 11 (3β-acetoxy-urs-12-en-28-oic acid) exhibited moderate inhibitory effects on Hela with IC₅₀ value of 55.0 and 68.8 µM, respectively. Moreover, 10 showed cytotoxic effect on HL-60 cell line with IC₅₀ value of 63.9 µM. To our knowledge, this is the first report that oleanderic acid was isolated from L. japonicum and investigated in cytotoxic effects on Hela and HL-60 cells.
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Humans , Antineoplastic Agents , Fruit , HL-60 Cells , Inhibitory Concentration 50 , Ligustrum , Mass Screening , Nerium , Oleaceae , Phytochemicals , Plants, MedicinalABSTRACT
Aim To evaluate the hypolipidemic effect of total phenylpropanoid glycoside extracted from Ligustrum robustum(Roxb.)Blume(CNTG)and its mecha-nisms.Methods The 60 hamsters were randomly divided into six groups,namely the control group,the model group,the positive control group(fenofibrate,150 mg·kg-1),the high(1 200 mg·kg-1),medium(600 mg·kg-1)and low(300 mg·kg-1)doses of CNTG groups.Only the control group was given control diet and other groups received high-fat diet.The changes of serum lipid were measured and analyzed in 1st week to ensure the successful establishment of the model.The drugs were administered daily for four weeks and the concentrations of lipids were determined in the 2nd week,3rd week and 4th week respectively.Quantitative real-time PCR and Western blot were used to assay the mRNA and protein expression of related signaling enzymes and proteins.Results Compared with the model group,the concentrations of serum TG,TC,LDL-C(P<0.05,P<0.01)and hepatic TG,TC(P<0.01)were effectively reduced in hamsters in CNTG-treated groups.Mechanism research found that CNTG increased the levels of phospho-AMPKα,LKB1 and phospho-LKB1 in liver(P<0.05).Conclusion CNTG prevents hyperlipidemia via activation of hepatic LKB1-AMPK pathway.
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Objective To identify the Ligustrum sinense Lour.and Jasminum elongatum(Bergium) Wild.from the macroscopic appearance and microscopic features.Methods Fresh samples of stems,leaves,flowers and fruits of the two kinds of medicinal plants were harvested.Stereoscopy was used for the observation of macroscopic appearance of Ligustrum sinense Lour.and Jasminum elongatum (Bergium) Wild.,and the microscopy was used for the examination of their microscopic features of stem and leaf cross section,tear film and the powder of leaf upper and lower epidermis.Results Original plant characteristics of the two kinds of species were as follows:Ligustrum sinense Lour.had a racemes and funnel-shaped corolla,while Jasminum elongatum (Bergium)Wild.had a cymes and salverform corolla.Microscopic identification results were as follows:the stem pith of Ligustrum sinense Lour.was smaller,and the stem pith ofJasminum elongatum(Bergium) Wild.was bigger;Ligustrum sinense Lour.stem powder had less pit or texture in the sclereids,and had reticulate vessels,and Jasminum elongatum(Bergium) Wild.stem powder had apparent pit or texture in the sclereids,and had spiral vessels.Conclusion The results will provide a basis for the identification,exploitation and utilization of Ligustrum sinense Lour.and Jasminum elongatum(Bergium) Wild.
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OBJECTIVE: To explore the active ingredients of wine fried Ligustrum lucidum fructus by studying the relationship between the HPLC chromatogram and antioxidant activity. METHODS: The characteristic chromatogram of wine fried Ligustrum lucidum fructus processed by means of classical homothermal acceleration was established by HPLC. DPPH, ABTS, and FRAP methods were established to determine the antioxidant activity. The spectrum-effect relationship was studied by using partial least squares regression (PLSR), and the chromatographic peaks related to antioxidant activity were identified. RESULTS: Peaks 1, 3, 8, 10, 14 and 15 were found to have positive relationship with DPPH free radical and ABTS free radical scavenging activity among the 17 matching characteristic chromatograms. Peaks 3, 8, 12, and 13 were significantly positively related to Fe3+ resuction capacity. Among the chromatographic peaks, peak 3 and peak 8 were positively related to the activities of scavenging DPPH free radical and ABTS free radical and reducing Fe3+. Peaks 3, 12, 13, 14, 15, and 17 were determined as salidroside, luteolin-7-O-glucoside, specnuezhenide, oleuropein, ligustroflavone, and luteolin, respectively. CONCLUSION: The pharmcodynamic effects of wine fried Ligustrum lucidum fructus do not depend on the contents of several index components such as specnuezhenide and salidroside. The quality of traditional Chinese medicines should be represented by the compound groups associated with pharmcodynamic effect.
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Objective To optimize the extraction and purification process of oleanolic acid in Ligustrum lucidum fruits with macroporous resin. Methods Grinding degree, temperature, liquid ratio, extraction time, extraction times, ethanol volume fraction and pH were selected as factors, which may influence the extraction of oleanolic acid from L. lucidum. Single factor experiments were carried out on these factors. The optimal extraction conditions were determined, and the best type of resin was chosen for separation and purification of oleanolic acid in the optimal conditions. Results The optimal technological conditions were as follows: the temperature was controlled at 80°C, ethanol solution was 60%, liquid material ratio was 1:11, extraction time was 120 min, and extraction times were twice. The crude extraction was processed at pH 12, and then the pH was adjusted to 2 to further purify the processed crude extraction. The content of oleanolic acid in the extract was 9.5% by UV visible spectrophotometric method. This extraction was further separated and purified by AB- 8 resin, and the content of oleanolic acid detected by ultraviolet visible spectrophotometr was 15.2%o Conclusion The optimal process of the oleanolic acid extraction is reproducible, simple and feasible. It may provide reference for using macroporous resin to extract oleanolic acid from L. lucidum in the industry production.
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Objective To optimize the extraction and purification process of oleanolic acid in Ligustrum lucidum fruits with macroporous resin. Methods Grinding degree,temperature,liquid ratio,extraction time,extraction times,ethanol volume fraction and pH were selected as factors,which may influence the extraction of oleanolic acid from L. lucidum. Single factor experiments were carried out on these factors. The optimal extraction conditions were determined,and the best type of resin was chosen for separation and purification of oleanolic acid in the optimal conditions. Results The optimal technological conditions were as follows:the temper?ature was controlled at 80℃,ethanol solution was 60%,liquid material ratio was 1∶11,extraction time was 120 min,and extraction times were twice. The crude extraction was processed at pH 12,and then the pH was adjusted to 2 to further purify the processed crude extraction. The content of oleanolic acid in the extract was 9.5%by UV visible spectrophotometric method. This extraction was further separated and purified by AB-8 resin,and the content of oleanolic acid detected by ultraviolet visible spectrophotometr was 15.2%.Conclusion The optimal process of the oleanolic acid extraction is reproducible,simple and feasible. It may provide refer?ence for using macroporous resin to extract oleanolic acid from L. lucidum in the industry production.
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Objective:To analyse the physicochemical properties and structure of major privet pollen allergen Lig v 1 using bioinformatics software and provide a reference for choosing suitable recombinant expression system for Lig v 1 and modifying the allergen Lig v 1 experimentally.Methods:The physicochemical properties were analysed by ProtParam,the signal peptide by SignalP 4.1 Server,the transmembrane helix by TMHMM Server v.2.0,the secondary structure by GOR4,MHCⅡepitopes by NetMHCⅡ2.2 Server,B-cell epitopes by ProteanTM 5.01 and the phylogenetic tree by MEGA 6.Results: Privet major pollen allergen Lig v 1 was stable in Escherichia coli and it doesn′t possess any signal peptide and transmembrane helix.Most secondary structures of Lig v 1 were random coils.Potential region of MHCⅡepitope of Lig v 1 was 30-44.Potential B-cell epitopes possess discontinuous and continuous a mino acid sequences.Lig v 1 and its counterparts from Fraxinus excelsior and Olea europaea were clustered into one group.Conclusion:Escherichia coli is the suitable expression system for recombinant Lig v 1.In silico prediction of the epitopes of Lig v 1 provides a reference for modifying the allergen Lig v 1 experimentally.
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OBJECTIVE:To establish a method for the contents determination of 4 ingredients in Ligustrum lucidum decoration piece by reference extract method. METHODS:HPLC was performed on the column of XBridge C18 with mobile phase of acetoni-trile-0.1% formic acid(gradient elution) at flow rate of 1.0 ml/min,detection wavelength was 254 nm,column temperature was 30℃and volume injection was 10μl. RESULTS:The linear range was 0.037-0.598 mg for neonuezhenide,0.006-0.101 mg for ac-teoside,0.189-3.023 mg for nuezhenide and 0.314-5.027 mg for specnuezhenide(r≥0.999 0);RSDs of precision,reproducibility and stability tests were no more than 3.8%;recoveries were 98.46%-104.83%(RSD=2.43,n=6),95.55%-104.57%(RSD=3.63, n=6),100.09%-104.39%(RSD=1.45,n=6)and 98.84%-104.97%(RSD=2.02,n=6). CONCLUSIONS:The method is simple, good reproducibility,and can be used for the contents determination of 4 ingredients in L. lucidum decoration piece.