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Objective:To investigate the optimal dose of Th1/Th2 immuno-enhancement effects of cultivated Artemisia rupestris L. crude polysaccharides (CARCP) on foot-and-mouth disease vaccine (FMDV) via the intramuscular route. Methods:ICR mice were intramuscularly immunized twice with different concentrations of CARCP mixed with FMDV at 2-week intervals. FMDV-specific antibodies, isotypes, and IgE in serum were detected by ELISA. Splenocyte proliferation was detected by MTT. T lymphocyte subsets and cytokines in the spleen were detected by flow cytometry. Clinical signs and local reactions at the injection site were monitored daily, and the body weight of mice was weighed after immunization.Results:The medium dose of CARCP could significantly improve FMDV-specific IgG, IgG 1, and IgG 2a antibody levels and the IgG 2a/IgG 1 ratio ( P<0.05) and lead to significant splenocyte proliferative responses ( P<0.01). The medium dose of CARCP could also significantly increase the level of CD3 +CD4 + and CD3 +CD8 + T cells as well as CD4 +/CD8 + ratio ( P<0.05), elicited the higher levels of IFN-γ in CD4 + T cells and CD8 + T cells ( P<0.05). No local adverse reactions at the injection site were observed after immunization. There was no significant difference in body weight or growth between each group( P>0.05). CARCP did not significantly induce an IgE response ( P>0.05). Conclusions:CARCP as an FMDV adjuvant promotes Th1/Th2 immune responses, especially in favor of the Th1 response, and has a certain safety profile. The best immune enhancement is achieved with CARCP at medium doses.
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Objective: To investigate the chemical constituents of the whole plants of Artemisia rupestris. Methods: The chemical constituents were isolated and purified by silica gel, Sephadex LH-20 column chromatography and semi-preparative HPLC. The structures of the isolated compounds were identified by NMR and MS spectroscopic method. Results: Eighteen compounds were isolated and purified, which structures were identified as 2α-(2',4'-hexadiynoyl)-1,6-dioxaspiro [4,5]-deca-3-ene (1), chrysophanol (2), ethyl vanillate (3), methyl linolenate (4), 8,11-octadecadienoic acid methyl ester (5), eicosane coumaric acid ester (6), emodin (7), 2,5-di-tert-butylphenol (8), phellopterin (9), syringic acid (10), 7-methoxycoumarin (11), vanillin (12), imperatorin (13), oroxylin A (14), 5-hydroxy-3,4',6,7-tetramethoxyflavone (15), erucylamide (16), octadecyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl) propanoate (17) and 3,5,3',4'-tetrahydroxy-6,7-dimethoxyflavone (18). Conclusion: Compounds 1, 4, 11-14 and 18 are isolated from A. rupestris for the first time, and all the other compounds were isolated from the genus Artemisia for the first time.
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@#To establish HPLC dual-wavelength fingerprint of Artemisia rupestris L. , and provide a scientific basis for the improvement of its quality specifications. The separation was performed on an Agilent Zorbax SB-C18(4. 6 mm×250 mm, 5 μm)column maintained at 32 ℃, with methanol-0. 2% formic acid-water gradient elution at flow rate of 1. 0 mL/min and the UV detection wavelength at both 245 and 325 nm. The sample injection volume was 20 μL. The fingerprint of Artemisia rupestris L. was established. 7 and 8 common peaks were found, respectively, of which 5 common peaks were identified, and the similarity among 9 batches of Artemisia rupestris L. and the fingerprints of control was over 0. 9. The HPLC dual-wavelength fingerprint of Artemisia rupestris L. was established for the first time, providing a new scientific basis for its identification and quality control.
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Objective@#To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L. crude polysaccharides (WARCP) as an adjuvant on ovalbumin (OVA) vaccine in mice immunized intramuscularly.@*Methods@#ICR mice were randomly divided into 6 groups (5 per group), including 9 g/L NaCl group (blank control), OVA group (10 μg OVA), low dose WARCP/OVA group (OVA+50 μg WARCP), medium dose WARCP/OVA group (OVA+200 μg WARCP), high dose WARCP/OVA group (OVA+400 μg WARCP), and aluminum adjuvant (Alum)/OVA group (positive control group, OVA+100 μg Alum). ICR mice were immunized intramuscularly and weighted. The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay (ELISA). T cells subsets from spleen and lymph nodes were detected by flow cytometry.@*Results@#The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels (all P<0.05). The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels (all P<0.05), but the difference was not statistically significant comparing with Alum/OVA group (P>0.05). The low-dose WARCP/OVA group enhanced the percentage of CD4+ T cells in spleen and CD4+ T, CD8+ T, CD4+CD44+ T cells in lymph nodes (all P<0.05). The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4+ T, CD8+ T, CD4+CD44+ T, CD8+CD44+ T cells in spleen and CD8+CD44+ T cell in lymph nodes (all P<0.05).@*Conclusions@#Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity, and it is safe and reliable. The results in this study provide a theoretical basis for the popularization and application of WARCP.
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Objective To investigate the enhancement effect of Xinjiang wild Artemisia rupestris L. crude polysaccharides (WARCP) as an adjuvant on ovalbumin (OVA) vaccine in mice immunized intramuscularly. Methods ICR mice were randomly divided into 6 groups (5 per group), including 9 g/L NaCl group (blank control), OVA group (10 μg OVA), low dose WARCP/OVA group (OVA+50 μg WARCP), medium dose WARCP/OVA group (OVA+200 μg WARCP), high dose WARCP/OVA group (OVA+400 μg WARCP), and aluminum adjuvant (Alum)/OVA group (positive control group, OVA +100 μg Alum). ICR mice were immunized intramuscularly and weighted. The OVA-specific antibodies and subtypes in serum were detected by enzyme linked immunosorbent assay (ELISA). T cells subsets from spleen and lymph nodes were detected by flow cytometry. Results The medium-dose WARCP/OVA group enhanced IgG and IgG1 levels and increased early antibody levels (all P<0.05). The medium-dose WARCP/OVA group and the high-dose WARCP/OVA group significantly enhanced IgG2a levels (all P<0.05), but the difference was not statistically significant comparing with Alum/OVA group (P>0.05). The low-dose WARCP/OVA group enhanced the percentage of CD4+ T cells in spleen and CD4 + T, CD8+ T, CD4 +CD44 + T cells in lymph nodes (all P<0.05). The medium dose WARCP/OVA group and the high dose WARCP/OVA group enhanced the CD4 + T, CD8 + T, CD4 +CD44 + T, CD8 +CD44+ T cells in spleen and CD8+CD44+ T cell in lymph nodes (all P<0.05). Conclusions Plant-derived WARCP as an OVA protein vaccine adjuvant can enhance cellular immunity and humoral immunity, and it is safe and reliable. The results in this study provide a theoretical basis for the popularization and application of WARCP.
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Due to the diversity and complexity, the change of chemical components in medicinal plant according to the time, cultivated varieties or ecological condition is difficult to recognize using traditional phytochemistry method. In order to analyze the pharmacodynamics material basis in Uighur medicinal plant Artemisia rupestris L. in an effective and comprehensive way, a plant metabolomics approach was established based on liquid chromatography-tandem mass spectrometry (LC-MS/MS). This study firstly focused on the effect of extraction solvents,redissolve solvents and ultrasonic time on the untargeted metabolomics, then the optimal preparation condition was selected according to metabolites coverage. After methodology validation, the approach was applied to acquire metabolic information in root, stem, branchlet, leaf and flower of Artemisia rupestris L. The results showed that the metabolome in flower was obviously different with the other organs. Coupling with multivariate statistical analysis, a batch of differential metabolites were picked out, in which 61 flavonoids, 97 rupestonic acid derivatives, 7 chlorogenic acids and 15 other compounds were primarily recognized according to the characteristic fragmentation rules of specific structure type and database retrieval. Additionally,the distribution characteristics of the above 180 differential metabolites was illustrated by cluster heat map. In conclusion,this study provided important information about the rational utilization of effective parts from Artemisia rupestris L.,and offered a novel strategy for quality control,variety improvement and reasonable development of medicinal plants.
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Objective To investigate the efficacy of using crude polysaccharides extracted from cultivated Artemisia rupestris L. ( CARCP) in Xinjiang as an immunologic adjuvant for ovalbumin ( OVA) . Methods The mice were subcutaneously immunized twice with OVA vaccine formulated with CARCP. ELISA assay was performed to measure the levels of IgG, IgG1 and IgG2a antibodies. Flow cytometry analy-sis was performed to detect the percentages of different T lymphocyte subsets ( CD3+CD4+ and CD3+CD8+T cells, CD4+CD44+ and CD8+CD44+T cells) in splenocytes as well as the expression of intracellular cyto-kines ( CD4+IFN-γ, CD8+IFN-γ) and CD4+CD25+Foxp3+Treg cells. Results The levels of IgG, IgG1 and IgG2a antibodies in serum, the percentages of CD3+CD4+, CD3+CD8+, CD4+CD44+ and CD8+CD44+T cells as well as the ratio of CD4+/CD8+T lymphocytes increased significantly in mice immunized with OVA in combination with CARCP (P<0. 05). Moreover, CARCP enhanced the expression of CD4+IFN-γand CD8+IFN-γ( P<0. 05 ) , but inhibited the expression of CD 4+CD 2 5+Foxp 3+Treg cells ( P<0. 05 ) . Conclusion As an adjuvant for OVA, CARCP enhanced the humoral and cellular immune responses, especially the T cell immune responses.
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Objective To investigate the efficacy of using Xinjiang wild Artemisia rupestris L. crude polysaccharides ( WARCP) as an immunologic adjuvant for influenza virus vaccine( IVV) .Methods ICR mice were subcutaneously immunized with 0.3 μg of IVV and 1.5 μg of IVV alone or co-administered with 200 μg of WARCP on 0 d and 14 d.Antibody levels in serum samples were detected by using indirect ELISA.MTT method was used to measure the proliferation of splenocytes.The growth conditions of mice were observed as well.Results No significant differences in the body weight were observed between mice from different groups (P>0.05).The levels of influenza virus-specific IgG, IgG1 and IgG2a were signifi-cantly increased in mice injected with WARCP adjuvant (P<0.05).The levels of IgG antibody in mice im-munized with low-dose of IVV and WARCP were significantly higher than those in mice immunized with high-dose of IVV alone (P<0.05), indicating at least 80% reduction in vaccine dosage by adding WARCP as adjuvant.Moreover, WARCP significantly promoted the proliferation of lymphocytes (P<0.05).Conclu-sion Adding WARCP to IVV enhanced the efficacy of IVV by boosting humoral and cellular immunity re-sponses with the advantages of high safety and dose-sparing.This study suggested the possibility of using WARCP as a novel immunologic adjuvant for influenza virus vaccine.
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Objective To establish a preparative method for rupestonic acid reference substance from Artemisia rupestris by HPLC.Methods After raw material being extracted with 95% ethanol,the ethanol extracts were evaporated to dryness under reduced pressure.Then the dry powder was extracted by ethyl acetate,the ethyl acetate extract was fractionated by column chromatography on silica gel(100—200 mesh) using petroleumbenzine-ethyl acetate(3∶1) as the eluent.The collected fraction was purified by preparative HPLC.Results The purity of the product quantitated by normality was over 98%.Conclusion The developed method is simple and rapid and the product of rupestonic acid can be used as reference substance.