Effects of acetyl groups on the prebiotic properties of glucomannan extracted from Artemisia sphaerocephala Krasch seeds.

This study explores the structural modification of glucomannan extracted from Artemisia sphaerocephala Krasch seeds (60S) to assess the impact of acetyl groups on its prebiotic characteristics. The structural changes were examined, with a focus on the degree of acetyl group substitution (DS). Both deacetylation and acetylation had limited influence on the molecular properties of 60S. Despite these modifications, the apparent viscosity of all samples remained consistently low. In vitro fermentation experiments revealed that Escherichia-Shigella decreased as DS increased, while Bacteroides ovatus was enriched. Acetylation had no significant impact on the utilization rate of 60S but led to a reduction in the production of propionic acid. Furthermore, untargeted metabolomics analysis confirmed the changes in propionic acid levels. Notably, metabolites such as N-acetyl-L-tyrosine, γ-muricholic acid, and taurocholate were upregulated by acetylated derivatives. Overall, acetyl groups are speculated to play a pivotal role in the prebiotic properties of 60S.

Facile fabrication of Artemisia sphaerocephala krasch gum hydrogels by radiation induced cross-linking polymerization and enhanced ultrahigh adsorption for methylene blue.

Although Artemisia sphaerocephala krasch gum (ASKG) has attracted growing attention in the field of medical engineering and food industries, however, there are few studies on the gelation of ASKG. In this paper, acrylic acid modified ASKG hydrogels were prepared by radiation induced grafting, cross-linking and polymerization technique for the first time. The semi-IPN structure was prepared by the cross-linked ASKG network and poly-AAc dispersed within the network. The effects of the adsorbed dose on the swelling ratio and gel fraction were investigated. The different acrylic acid content modified ASKG hydrogels (ASKGAAc1 and ASKGAAc2) for methyl blue (MB) adsorption were investigated, and the ASKG hydrogels was also studied for comparison. The influence of pH, contact time, initial concentration, temperature, ion strength on MB adsorption were tested. The results showed that acrylic acid can promote the formation of hydrogel and greatly enhanced the adsorption of ASKG. The adsorption isotherms were well obeyed the Langmuir model, and the maximum adsorption capacity for MB of ASKG, ASKGAAc1 and ASKGAAc2 were 571.43, 1517.8 and 1654.9 mg/g, respectively. Moreover, the MB adsorption by ASKG based hydrogels was exothermic, spontaneous, and more favorable at lower temperature. Furthermore, the adsorption-desorption experiments demonstrated a good reusability of these hydrogels.

Physicochemical properties and cytocompatibility of radiation-resistant and anti-washout calcium phosphate cement by introducing artemisia sphaerocephala krasch gum.

The anti-washout ability of calcium phosphate cement (CPC) determines the effectiveness of CPC in clinical application. The γ-ray irradiation method often used in the sterilization process of CPC products is easy to degrade some commonly polymer anti-washout agent, which greatly reduces its anti-washout performance. Artemisia sphaerocephala Krasch gum (ASKG) has the potential of radiation resistance and anti-washout, but no one has considered its performance as anti-washout agent of CPC and mechanism of radiation resistance and anti-washout so far. In this study, we report the effect of γ-ray on ASKG and the effectiveness of ASKG for enhancing of radiation resistance and anti-washout ability of CPC, the physical, chemical properties and in vitro cell behaviors of ASKG-CPCs were also investigated. The results showed that addition of ASKG before and after irradiation could significantly enhanced the anti-washout performance of CPC, which is differ from conventional anti-washout agents. Meanwhile, ASKG-CPCs had an excellent injectable property and biocompatibility, and low content of irradiated ASKG could promote bone differentiation well. We anticipate that the radiation-resistant and anti-washout ASKG-CPCs have potential application prospect in orthopaedic surgery.

Effect of Artemisia sphaerocephala krasch gum on the functional properties of pork batters.

The influence of the incorporation of Artemisia sphaerocephala krasch gum (ASK gum; 0-0.18%) on the water holding capability (WHC), texture, color, rheological property, water distribution, protein conformation and microstructure of pork batters was investigated. The results showed that the cooking yield, WHC and L* value of pork batter gels significantly increased (p < .05) with the increasing incorporation of ASK gum, and the highest value were observed at 0.15%; the a* value decreased significantly (p < .05) and no significance was obtained in b* value (p > .05); the hardness, elasticity, cohesiveness and chewiness increased first and then decreased, and reached the highest value at 0.15%. The rheological results showed that the higher G' value was obtained in pork batters by the incorporation of ASK gum; the low field NMR analysis indicated that ASK gum significantly increased the proportion of P2b and P21 (p < .05) and decreased the proportion of P22 ; Fourier transform infrared spectroscopy (FTIR) indicated that the ASK gum significantly reduced the α-helix content and increased the ß-sheet content (p < .05). Scanning electron microscopy results suggested that the incorporation of ASK gum could promote the formation of a more homogeneous and stable microstructure of pork batter gels. Therefore, appropriate incorporation (0.15%) of ASK gum may improve the gel properties of pork batters, and while excessive incorporation (0.18%) could weaken the gel properties.

Stability improvement of emulsion gel fabricated by Artemisia sphaerocephala Krasch. polysaccharide fractions.

Artemisia sphaerocephala Krasch. polysaccharide (ASKP) contained two fractions of 60P and 60S with different molecular weight. It was found the potential performance of interface adsorption and gelation activities for the high molecular weight of 60P in comparison with low molecular weight of 60S. The emulsion stability and droplets filling in gel network was highly dependent on the medium chain triglyceride (MCT) concentrations. The emulsion gels fabricated through a complexation of 60P and gelatin or collagen peptides exhibited significantly improved emulsifying activity and gel strength at higher concentration of MCT. Gelatin or collagen peptide could be adsorbed on the droplets interface and interact with 60P in gel matrix, thus presenting an active filling. However, 60P based emulsion gel complexed with pullulan contributed to a lower strength than hydrogel, which was probably due to the existence of spaces between droplets and gel matrix, weakening the stability of gel network, considered as an inactive filling.

Construction of Artemisia sphaerocephala Krasch. Polysaccharide based hydrogel complexed with pullulan and gelatin crosslinked by ferric ions.

Artemisia sphaerocephala Krasch. polysaccharide (ASKP) was found to be crosslinked with ferric ions to form hydrogels in the previous study. In this work, it was demonstrated that ASKP-Fe3+ hydrogel complexed with pullulan or gelatin contributed to a significantly enhanced gel strength at 1.5% ASKP, 60 mM Fe2+, pH 4.0, and the mixing ratio of 9: 1. The complexed hydrogels presented a dense semi-interpenetrating network along with the delay of gelation time and the increase of water retention. ASKP based complexes exhibited good compatibility, probably because pullulan and gelatin could be entangled with ASKP chain under hydrogen bonding and electrostatic interaction, respectively. The interaction between ASKP and pullulan or gelatin contributed to the formation of complexed hydrogels with dense network and significantly enhanced gel strength. It is inferred that ASKP would have great potential to be a new gelling material as well as for the ferric ions delivery.

Transcriptomic Analysis Reveals Key Genes Involved in Oil and Linoleic Acid Biosynthesis during Artemisia sphaerocephala Seed Development.

Artemisia sphaerocephala seeds are rich in polysaccharides and linoleic acid (C18:2), which have been widely used as traditional medicine and to improve food quality. The accumulation patterns and molecular regulatory mechanisms of polysaccharides during A. sphaerocephala seed development have been studied. However, the related research on seed oil and C18:2 remain unclear. For this study, A. sphaerocephala seeds at seven different development stages at 10, 20, 30, 40, 50, 60, and 70 days after flowering (designated as S1~S7), respectively, were employed as experimental samples, the accumulation patterns of oil and fatty acids (FA) and the underlying molecular regulatory mechanisms were analyzed. The results revealed that oil content increased from 10.1% to 20.0% in the early stages of seed development (S1~S2), and up to 32.0% in mature seeds, of which C18:2 accounted for 80.6% of the total FA. FA and triacylglycerol biosynthesis-related genes jointly involved in the rapid accumulation of oil in S1~S2. Weighted gene co-expression network analysis showed that transcription factors FUS3 and bHLH played a critical role in the seed oil biosynthesis. The perfect harmonization of the high expression of FAD2 with the extremely low expression of FAD3 regulated the accumulation of C18:2. This study uncovered the gene involved in oil biosynthesis and molecular regulatory mechanisms of high C18:2 accumulation in A. sphaerocephala seeds; thus, advancing research into unsaturated fatty acid metabolism in plants while generating valuable genetic resources for optimal C18:2 breeding.

The anti-obesity effects exerted by different fractions of Artemisia sphaerocephala Krasch polysaccharide in diet-induced obese mice.

Artemisia sphaerocephala Krasch polysaccharide (ASKP) consists of two main fractions, 60P (molecular weight at 551 kDa) and 60S (molecular weight at 39 kDa). The anti-obesity effects of ASKP and its two fractions were investigated in high-fat-diet-fed mice and showed similar capability in efficiently preventing the development of obesity. The final body weight and body weight gain of obesity mice model were reduced by 12.44% and 35.33% by ASKP, 10.63% and 34.35% by 60P, and 7.82% and 20.04% by 60S. They also showed similar efficiency to ameliorate dyslipidemia, systematic inflammation, and gut dysbiosis. The colonic genes of barrier integrity were significantly upregulated and the genes of hepatic lipid metabolism and that of colonic inflammatory response were suppressed. They attenuated the gut dysbiosis in obese mice, such as the significant enrichment of beneficial genera (Bifidobacterium and Olsenella) and suppression of harmful ones (Mucispirillum and Helicobacter). Significant enrichment of carbohydrate metabolism associated with the promotion of short-chain fatty acid production and decrease of the metabolisms related to obesity and gut dysbiosis (valine, leucine, and isoleucine biosynthesis, and nitrogen metabolism) were also observed by the administration of ASKP, 60P, and 60S. Overall, these polysaccharides showed potential in acting as prebiotics in preventing high-fat-diet-induced obesity.

Transcriptome and co-expression network analysis reveal molecular mechanisms of mucilage formation during seed development in Artemisia sphaerocephala.

Seed mucilage has significant economic value. However, the identification of key regulatory genes in mucilage formation and their molecular regulatory mechanism remain unknown. Artemisia sphaerocephala seeds are rich in mucilage. In this study, A. sphaerocephala seeds in 10, 20, 30, 40, 50, 60 and 70 days after flowering were used as materials to reveal their molecular regulatory mechanism in mucilage formation by RNA-sequencing and weighted gene co-expression network analysis (WGCNA). 21 key regulatory genes for mucilage formation were identified, including AsKNAT7 and AsTTG1 genes, as well as AsNAM and AsAP2 gene families. From 10-30 days after flowering, both AsNAM and AsAP2 supported mucilage formation. From 40-70 days after flowering, promotion by AsNAM and AsAP2 was weakened and the up-regulation of AsKNAT7 inhibited mucilage formation, leading to no further increases in mucilage content. This in depth elucidation of seed mucilage formation lays the foundation for the application of mucilage.

Synthesis of Se-polysaccharide mediated by selenium oxychloride: Structure features and antiproliferative activity.

Selenium oxychloride (SOC) was employed as a highly reactive selenide reagent to synthesize selenized Artemisia sphaerocephala polysaccharides (SeASP). Se content of SeASP was significantly increased (∼22,400 µg/g) as compared to HNO3/H2SeO3 selenylation method (1703 µg/g). Furthermore, selenized ASP was prepared by using microwave-assisted synthesis which obviously enhanced selenylation kinetics. FT-IR, Raman, XPS and NMR results exhibited seleno-group was substituted at C6 position in the form of selenite (Se4+). SEC-MALLS suggested SOC system could effectively avoid the degradation of polysaccharide chain. Meanwhile, MALLS calculation, MB spectrophotometric method and AFM observation showed SeASP appeared spherical and rod-shaped conformation after selenylation. Seleno-groups were more likely to affect the conformational transformation of polysaccharide chains. Moreover, SeASP could significantly enhance antiproliferative activity against three tumor cells, of which the IC50 value of HepG2 was calculated as 24.35 µg/mL. It was found that higher Se content could effectively improve the antitumor activities of Se-polysaccharides in vitro.

[HNMP]HSO4 catalyzed synthesis of selenized polysaccharide and its immunomodulatory effect on RAW264.7 cells via MAPKs pathway.

In this paper, selenized Artemisia sphaerocephala polysaccharides (SePAS) were obtained through employing N-methyl-2-pyrrolidone hydrosulfate as catalyst, which showed a maximum Se content enhanced to 8744 µg/g. FT-IR, 1D/2D NMR, X-ray photoelectron spectroscopy (XPS) and size-exclusion chromatograph analysis exhibited that Se had been successfully introduced into PAS and existed in the form of selenate group (Se4+) with the substitution position at C-6. Furthermore, immunostimulating assays indicated that SePAS with high Se content exhibited stronger immunomodulatory activities by upregulated the phosphorylation level of ERK, JNK and p38, thus enhancing RAW264.7 cells proliferation, phagocytosis, levels of interleukin-6, nitric oxide, tumor necrosis factor and interleukin-1ß. The current outcome suggested that Se content might be a critical factor affecting the immunomodulatory effects of selenized PAS on macrophage RAW264.7.

Prebiotic properties of different polysaccharide fractions from Artemisia sphaerocephala Krasch seeds evaluated by simulated digestion and in vitro fermentation by human fecal microbiota.

Artemisia sphaerocephala Krasch polysaccharide (ASKP) and its two fractions-60P (branched xylan) and 60S (branched glucomannan), were subjected to simulated gastrointestinal digestion and in vitro fermentation by human fecal microbiota. The results showed that all polysaccharide fractions could transit through gastrointestinal tract without dramatic degradation and be utilized by gut microbiota. ASKP exhibited the highest depletion rate and highest capability to decrease the pH than its fractions. Meanwhile, 60S showed the stronger capability to increase the production of propionic acid and reduce the ratio of acetic acid to propionic acid. At the phylum level, all polysaccharides efficiently reduced the Firmicutes/Bacteroidetes ratio and relative abundance of Proteobacteria, with ASKP being the most capable to suppress the proliferation of Proteobacteria. At the genus level, ASKP and 60P markedly promoted the growth of Bacteroidetes, and 60S promoted the growth of Parabacteroides and Collinsella. Prediction on metabolic function revealed that polysaccharide administration could dramatically change the metabolic profile of bacteria compared with fructooligosaccharides. Besides, all the polysaccharides dramatically promoted the bile acid metabolism. Compared with 60S, ASKP and 60P showed stronger ability to suppress the metabolisms on carbohydrate and amino acid. In summary, both ASKP and its two fractions showed the prebiotic potentials.

Controlling dough rheology and structural characteristics of chickpea-wheat composite flour-based noodles with different levels of Artemisia sphaerocephala Krasch. gum addition.

The effect of Artemisia sphaerocephala Krasch. gum (ASKG) addition on the dough performance and structural characteristics of the wheat-chickpea composite flour-based product was studied. The low levels of ASKG addition (0.03-0.5%) tended to cause a remarkable increase in the viscoelastic properties of the composite dough system, followed by a decreased trend at higher level of gum addition (0.8%). This trend was in agreement with the variations found for cooking loss, free -SH content, and relative crystallinity measured by X-ray diffractometer for dough samples. The confocal laser scanning micrographs (CLSM) further confirmed the above trend. For cooked noodle samples, the variations in pasting properties was supported by the changes in relatively crystallinity and free sulfhydryl content, suggesting a relatively more compact structure was formed at the 0.3% of gum addition. The scanning electron microscopic and CLSM observations both revealed that a relatively denser and more coherent network structure was achieved at 0.3% ASKG addition. On the other hand, the higher levels of gum substitution at 0.5% and 0.8% tended to disrupt this stronger network with visible signs of starch deformation, due to the inefficient entrapment of starch molecules during cooking as a result of the lack of gluten network.

Trivalent iron induced gelation in Artemisia sphaerocephala Krasch. polysaccharide.

Artemisia sphaerocephala Krasch. polysaccharide (ASKP) has attracted growing attention in the field of food and medical engineering due to its biological activity and colloidal property. In this study, the binding between ASKP and ferric ions was found and the binding mechanism was explored. The results showed that ASKP could form a hydrogel with three-dimensional network structure in the presence of ferric ions. Ferric ions could specifically bind with the carboxyl and hydroxyl groups of the high molecular weight fraction of 60P with the binding stoichiometry of [M3+]/[repeating unit] = 2.5. The possible mechanism of the formation of ASKP-Fe3+ complex was proposed as two binding modes of monodentate and bridging binding. ASKP-Fe3+ complex exhibited higher thermal stability than ASKP revealed with DSC thermograms. The study indicated that ASKP would be a novel gelation biopolymer and the ASKP-Fe3+ complex hydrogel could be exploited as a new iron fortifier.

Artemisia sphaerocephala Krasch polysaccharide mediates lipid metabolism and metabolic endotoxaemia in associated with the modulation of gut microbiota in diet-induced obese mice.

Artemisia sphaerocephala Krasch polysaccharide (ASKP) has been proved to have many bioactivities. To determine the underlying mechanisms on anti-obesogenic effect of ASKP in mice, parameters related to obesity, gut microbiota composition, and the correlation between the parameters and specific bacterial taxa were investigated. The results showed that ASKP significantly alleviated high-fat-diet-induced obesity in mice with the amelioration of dyslipidemia, and metabolic endotoxaemia. Relative expression analyses of genes indicated that ASKP administration modulated hepatic lipid metabolism with the downregulation of related genes, including ACC-1, FAS, SREBP-1c, and PPARγ. 16S rRNA analysis showed that ASKP mediated the gut dysbiosis induced by high-fat diet, such as the reduction of Proteobacteria, AF12, and Helicobacter. Spearman's correlation showed that some specific genera, such as Odoribacter, AF12, and Rikenella, were strongly associated with obesity-related parameters. Our results demonstrated that ASKP could serve as a potential prebiotic agent in the prevention of diet-induced obesity.

Preparation, structural characterization and bioactivity of 4-O-Methylglucuronoxylan from Artemisia sphaerocephala Krasch.

We obtained four soluble acid xylan fractions AGP-III-A, AGP-III-B, AGP-III-C and AGP-III-D from the insoluble Artemisia sphaerocephala Krasch gum (ASKG) polysaccharide by weak alkali treatment combined with H2O2-Vc oxidative degradation. Activity studies showed that the degradation components could reduce the cell viability of several cancer cell lines in a concentration-dependent manner, especially 4-O-Methylglucuronoxylan AGP-III-C with specific molecular weight and branching degree significantly reduced cancer cells viability and induced HepG2 apoptosis, also caused mitochondrial membrane dysfunction upregulated ROS levels, and induced G0/G1 arrest in HepG2 cells by cell cycle assay. Further, AGP-III-C mediates apoptosis in HepG2 cells by upregulating MAPK phosphorylation. The structure of AGP-III-C was characterized by uronic acid reduction, permethylation with GC-MS, and 2D-NMR analysis. The structure of AGP-III-C had a linear (1→4)-linked ß-Xylf residue backbone with one branched 4-O-Me-α-GlcAp attached to the main chain by a (1→2)-glycosidic bond at every two ß-(1→4)-Xylf units.

Arbuscular mycorrhizal symbiosis and achene mucilage have independent functions in seedling growth of a desert shrub.

Arbuscular mycorrhizal (AM) symbiosis can play a role in improving seedling establishment in deserts, and it has been suggested that achene mucilage facilitates seedling establishment in sandy deserts and that mucilage biodegradation products may improve seedling growth. We aimed to determine if AM symbiosis interacts with achene mucilage in regulating seedling growth in sand dunes. Up to 20 A M fungal taxa colonized Artemisia sphaerocephala roots in the field, and mycorrhizal frequency and colonization intensity exhibited seasonal dynamics. In the greenhouse, total biomass of AM fungal-colonized plants decreased, whereas the root/shoot ratio increased. AM symbiosis resulted in increased concentrations of nutrients and chlorophyll and decreased concentrations of salicylic acid (SA) and abscisic acid (ABA). Achene mucilage had a weaker effect on biomass and on nutrient, chlorophyll, and phytohormone concentration than did AM symbiosis. We suggest that AM symbiosis and achene mucilage act independently in enhancing seedling establishment in sandy deserts.

Selection and validation of reference genes for quantitative real-time PCR in Artemisia sphaerocephala based on transcriptome sequence data.

Artemisia sphaerocephala, a dicotyledonous perennial semi-shrub belonging to the Artemisia genus of the Compositae family, is widely distributed in northwestern China. This shrub is one of the most important pioneer plants which is capable of protecting rangelands from wind erosion. It therefore plays a vital role in maintaining desert ecosystem stability. In addition, to its use as a forage grass, it has excellent prospective applications as a source of plant oil and as a plant-based fuel. The use of internal genes is the basis for accurately assessing Real time quantitative PCR. In this study, based on transcriptome data of A. sphaerocephala, we analyzed 21 candidate internal genes to determine the optimal internal genes in this shrub. The stabilities of candidate genes were evaluated in 16 samples of A. sphaerocephala. Finally, UBC9 and TIP41-like were determined as the optimal reference genes in A. sphaerocephala by Delta Ct and three various programs. There were GeNorm, NormFinder and BestKeeper.

Synthesis and structural features of phosphorylated Artemisia sphaerocephala polysaccharide.

Phosphorylation is a key route to achieve varieties of biological activities for polysaccharides. Currently, conventional phosphorylation reagents are characterized by long processing times and high reaction temperatures. Here, we reported phosphorylation of Artemisia sphaerocephala polysaccharides (PASP) with the degrees of substitution (DS) of 0.34-0.54 in mild reaction conditions (3-4h and 25°C) by employing POCl3/pyridine technique. FT-IR spectroscopy and X-ray photoelectron spectroscopy (XPS) analysis confirmed the appearance of phosphate ester (P5+) groups in PASP. Size exclusion chromatography combined with multi-angle laser light scattering (SEC-MALLS) result clearly showed temperature-dependent increment of weight average molecular mass (MW). High temperature facilitated the reaction of phosphoryl chloride with primary and secondary hydroxyl groups, favoring the crosslinking of polysaccharide chains by phosphate di-esters as further confirmed in 13C NMR determination. Our results provide insight into the relationship between the structure and reaction conditions of phosphorylated polysaccharide, which is important for understanding and exploiting these derivatives in a wide range of applications.

Sulfation can enhance antitumor activities of Artemisia sphaerocephala polysaccharide in vitro and vivo.

In this study, a sulfated Artemisia sphaerocephala polysaccharide (ASPs) was prepared and its antitumor activity was evaluated in tumor cells and Hepatoma 22 (H22) tumor-bearing mice. In vitro experiments, ASPs significantly inhibited the growth of HepG2 and Hela cells with the IC50 values of 172.03 and 161.42µg/mL, respectively. Moreover, no direct cytotoxicity against mouse fibroblast L929 normal cells was observed in vitro. After oral administration for 12days, the tumor growth was significantly suppressed by ASPs at the doses of 200mg/kg (inhibition rate of 60.85%). Results of tumor histological morphology and cell cycle analysis showed that ASPs could arrest H22 cells at S phase and promote cell apoptosis. Additionally, immunohistochemical analysis demonstrated that ASPs caused the down-regulation of mutant p53 protein expression in a dose-dependent manner. Therefore, these findings proposed new insight into antitumor properties of sulfated polysaccharide as a promising agent in cancer treatment.