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Proposed structures of polysaccharides from A. carmichaelii leaves. HG, homogalacturonan; RG-I, type I rhamnogalacturonan; AG-II, type II arabinogalactan; AG-I, type I arabinogalactan. Graphical symbols are depicted according to the symbol nomenclature for glycans (SNFG) (Varki et al., 2015).
Source publication
Roots of Aconitum carmichaelii are used in Asian countries due to its content of bioactive alkaloids. In the production of root preparations, tons of leaves are usually discarded, leading to a huge waste of herbal material. The aim of this study is to investigate the polysaccharides in these unutilized leaves. A neutral polysaccharide (AL-N) appear...
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... or XLXG model due to the ratio of relative amounts of T-α-Xyl and1,2-linked α-Xyl (7.7:4.7, Table 3). Given the homogenous composition observed in Fig. 2D and Fig. 2E, AL-N might be a mixture of mannans, xyloglucans and/or glucomannans and minor amounts of arabinogalactan with similar Mw, as depicted in Fig. 4. The rather low yield of this fraction compared to the high yield of AL-I (Table 1) was the reason for not perform in further studies on ...
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... the signals of H/C-atoms at δ 5.09/104.3, and weak ones at δ 5.11/101.8 and δ 5.02/100.6 in HSQC, they belong to anomeric H/C atoms of 1,4-α-GalpA (GA 1,4 ), 1,4-α-GalpA-6-O-Me (GA 1,4Me ) and 4-α-3-O-Ac-GalpA (GA* 1,4 ) respectively ( Patova et al., 2019;Shakhmatov et al., 2019;Zou et al., 2020). Peaks in the downfield region in 13 C NMR at 173.8, 177.1 and 177.6 ppm should be assigned to C-6 of GalpA. ...
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... Fig. 3D) and downfield areas (such as residues H1/C6-GA 1,4Me , H5/C6-GA 1,4Me and H5/C6-GA 1,4 in Fig. 3D) in HMBC. Most proton signals correlated with H-1 of GalpA were appointed to H-2 by cross peaks in COSY (Fig. S2C), and their correlations to C-1 of GalpA in HMBC (Fig. 3D). Carbon signals correlated to H-1 were assigned to C-2/3/4 of GalpA (Fig. S4D, a). Some of the 1,4-α-GalpA residues were O-6 methyl esterified. Because of the downfield shifts of H-5 from about 4.7 ppm to about 5.10 ppm and the Table 4 1 H and 13 C NMR chemical shifts (ppm a ) assignment of AL-I-I and ...
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... studies (Kostálová et al., 2013;Patova et al., 2019). Particularly, a 4 → β-GalpA was found in AL-I-II, since cross peaks of H/C at δ 4.59/98.8 (GA′-1), δ 4.38/80.1 (GA′-4) and δ 3.49/74.4 ppm (GA′-2) in HSQC, δ 4.06/98.8 (H5/C1-GA′), δ 3.49/98.8 (H2/C1-GA′), δ 4.06/ 176.7 (H5/C6-GA′) in HMBC (Fig. 3D) and H1/H2 and H2/H3 correlations in COSY (Fig. S4C) were detected, which also has been shown in other studies ( Patova et al., 2019;Patova et al., 2021;Zou et al., 2020). The β-linkage was detected in AL-I-II due to the high-resolution 800 MHz NMR instrument, and it might be the reason that this structure has not been highly mentioned in most papers related to pectins. The signals of ...
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... addition, the residues TR-1, TR-2, and TR-4 in HSQC demonstrated the presence of terminal α-Rhap, as well as H/C cross peaks at δ 1.29/ 71.9, δ 1.29/74.8 and δ 1.24/71.6 in HMBC (Fig. S4D, b) and H/H cross peak at δ 1.29/3.90 in COSY spectra (not shown), as described in earlier published studies (Cui et al., 2007;Makarova et al., 2016). Likewise, the terminal α-Rhap residue might be located at the end of GlcpA, Galp, ...
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... belonging to Araf and Galp were measured in methylation analysis, such as 1,2-, 1,3-, 1,3,5-linked Araf and 1,6-, 1,3,6-and 1,4,6-linked Galp. Thus, according to the aforementioned results and NMR elucidation, both AL-I-I and AL-I-II could be typical pectin polysaccharides with both methyl-and acetyl-esterified α-GalA units, as depicted in Fig. 4. According to the known structure of plant-derived pectic polysaccharides ( Kaczmarska et al., 2022;Zaitseva et al., 2020) and the results of glycosidic linkages and NMR analysis above, AL-I-I was probably mainly composed of AG-II and arabinan as side chains of a RG-I core chain besides a HG backbone. The correlations in NMR were ...
Citations
... Methylation analysis was performed on native polysaccharides that did not contain uronic acid polysaccharides or reduced polysaccharides [42,43]. The polysaccharide samples were analyzed by GC-MS (Thermo Fisher Scientific, USA) using dichloromethane as the solvent. ...
Polysaccharides are the main nutritional constituents in the Armillaria mellea fruiting bodies. The structure and immunoactivity of Armillaria mellea polysaccharide are valuable to be determined for development. In the present study, two polysaccharides, including Armillaria mellea neutral polysaccharide (AMPN) and Armillaria mellea acidic polysaccharide (AMPA), were prepared and determined. AMPN and AMPA were separated and refined by the ion exchange column and gel chromatographycolumn. Analysis of AMPN and AMPA revealed molecular weights of 4.432 × 103 Da and 7.323 × 103 Da. The monosaccharide composition analysis revealed that AMPN was mainly composed of 68.3%glucose, while AMPA consisted primarily of glucose, mannose, and galactose, accompanied by 8.9% galacturonic acid and 3% fucose. Then, infrared spectra analysis, Congo red staining, methylation, and NMR spectroscopy analysis were conducted as a means to clarify the structure of AMPN and AMPA. The NMR spectra demonstrated that the two polysaccharides included both α and β-configuration glycosidic bonds. The Congo red experiment suggests that AMPN and AMPA all had obvious triple helix structures.The effects of AMPN and AMPA on immune repair ability were compared by immune deficiency mice. The activity experiment showed that AMPN and acidic polysaccharides extracted from the Armillaria fruiting body have biological immune activity. Among them, AMPA showed higher immune activity. These findings suggest that Armillaria mellea fruiting bodies may be used as a source of dietary supplements and functional products.
... To date, the presence of pectic polysaccharides with a branched RGÀ I core and HG domains have rarely been reported in Aconitum plants, except for pectins found in A. carmichaelii leaves in our previous study. [35] The present study is the first to show the presence of pectins in the roots of A. septentrionale. ...
... The fractionation of the polysaccharide fractions was carried out as described previously, [35] as shown in Figure S1D. In short, ASP (764 mg) was fractionated by IEC using a column ( ...
... Monosaccharide composition of the isolated polysaccharide fractions was determined by methanolysis and GC according to a previous study with minor modification as earlier described. [35] Briefly, samples were subjected to acid methanolysis with HCl (3 M) in water-free methanol (24 h, 80°C). Trimethylsilylated derivatives of the methyl glycosides were analyzed by capillary gas chromatography on a Trace™ 1300 GC (Thermo Scientific, Milan, Italy) processed by Chromelion Software v.6.80 (Dionex Corporation, Sunnyvale, CA, USA). ...
Aconitum septentrionale is known to contain toxic diterpene alkaloids, while other bioactive compounds in the plant remain unclear. The aim of this study was to explore the phenolic compounds and polysaccharides from the water extract of A. septentrionale roots. Fifteen phenolic compounds were isolated and identified by NMR and MS, including fourteen known and one new dianthramide glucoside (benzoic acid, 2-[[2-(β-D-glucopyranosyloxy)-5-hydroxybenzoyl] amino]-4, 5-dihydroxy-, methyl ester, 14). One neutral (complex of glucans with minor amounts of mannans) and two acidic polysaccharide fractions (complexes of pectic polysaccharides and glucans) were also obtained. Hydroxytyrosol (1), hydroxytyrosol-1-O-β-glucoside (2) and bracteanolide A (7) inhibited the release of nitric oxide by dendritic cells. Magnoflorine (8) and 2-[[2-(β-D-glucopyranosyloxy)-5-hydroxybenzoyl] amino]-5-hydroxy-, methyl ester (12) inhibited 15-lipoxygenase, and bracteanolide A (7) was a moderate inhibitor of xanthine oxidase. This study is the first to describe the diversity of phenolics and polysaccharides from A. septentrionale and their anti-inflammatory and anti-oxidant activities.
... Na + Na + Na + Na + species are also well known as immune-stimulant agents and many reports indicated that polysaccharides in these species have significant immune-stimulant properties (Zhao et al 2006;Gao et al. 2010;Fu et al. 2022). FPS-1, a water-soluble polysaccharide isolated from roots of A. carmichaelii Debx., showed potent stimulating effects on splenocyte antibody production and murine lymphocyte proliferation induced by concanavalin A or lipopolysaccharide (Zhao et al 2006). ...
The genus Aconitum belongs to the family Ranunculaceae, is endowed with more than 350 species on the earth. Medicinally important aconitine type of diterpenoid alkaloids are the characteristic compounds in most of the Aconitum species. The present review endeavored the major research carried out in the field of genetic resource characterization, pharmacological properties, phytochemistry, major factors influencing quantity, biosynthetic pathways and processing methods for recovery of active ingredients, variety improvement, propagation methods, and important metabolite production through cell/organ culture of various Aconitum species. More than 450 derivatives of aconitine-type C19 and C20-diterpenoid alkaloids along with a few other non-alkaloidal compounds, such as phenylpropanoids, flavonoids, terpenoids, and fatty acids, have been identified in the genus. A few Aconitum species and their common diterpenoid alkaloid compounds are also well characterized for analgesic, inflammatory and cytotoxic properties. However, the different isolated compound needs to be validated for supporting other traditional therapeutical uses of the plant species. Aconitine alkaloids shared common biosynthesis pathway, but their diversification mechanism remains unexplored in the genus. Furthermore, the process needs to be developed on secondary metabolite recovery, mass-scale propagation methods, and agro-technologies for maintaining the quality of products. Many species are losing their existence in nature due to over-exploitation or anthropogenic factors; thus, temporal monitoring of the population status in its habitat, and suitable management programs for ascertaining conservation needs to be developed.
... The activities of polysaccharides can vary widely, even among those from the same plant, demonstrating a direct relationship between polysaccharide structure and their biological activity [154]. Given the complex structures and various types of polysaccharides, their separation and purification, as well as structure determination, can be very difficult [155], and the characteristics and structures of most reported active polysaccharides are unknown. Therefore, effective new technologies to analyze the relationship between polysaccharide structure and their therapeutic effects, as well as their mechanisms of action in CRC, warrant further exploration. ...
Colorectal cancer (CRC) ranks third and second among the most widespread cancers worldwide and the most common causes of human death due to cancer, respectively. Further, for unknown reasons, numbers of young patients diagnosed with colon cancer has increased. Polysaccharides are important functional phytochemicals reported to have anti-CRC effects. Moreover, CRC development and progression is closely related to the gut microbiome. Although approaches for treating CRC have been the subject of some review papers, research into traditional Chinese medicine (TCM) treatments for CRC and the underlying mechanisms involving polysaccharides have not been reviewed. Here, we reviewed the mechanisms underlying treatment of CRC using TCM polysaccharides, based on the etiology of CRC, and common treatment methods applied. The relationship between intestinal microbes and CRC, the mechanism by which TCM polysaccharides induce CRC cell apoptosis, and how TCM polysaccharides promote immune responses are discussed, as well as TCM polysaccharide use in combination with chemotherapy. TCM polysaccharides provide options for CRC treatment, due to their advantages of having multiple targets, eliciting modest adverse reactions, and wide range of available sources.
... To date, the aerial parts of A. carmichaelii have shown similar analgesic and anti-inflammatory activities as for the roots [4]. An acidic polysaccharide fraction composed of pectic polysaccharides isolated from A. carmichaelii leaves and structurally characterized in a previous study, was shown to possess immunomodulatory and intestinal anti-inflammatory activities in vitro [5]. ...
... 517 m. a.s.l.). The fresh leaves were separated and processed as earlier reported [5]. ...
... A major acidic polysaccharide fraction, AL-I, was isolated from the water extract of dried leaves of A. carmichaelii by anion exchange chromatography, as earlier described [5]. The monosaccharide composition of AL-I was determined as trimethylsilylated (TMS) derivatives, after methanolysis by GC (Trace™ 1300 GC, Thermo Scientific TM, Milan, Italy), as described by Chambers and Clamp [13] with modifications [14]. ...
The industrial processing of Aconitum carmichaelii roots for use in Traditional Chinese Medicine generates a high amount of waste material, especially leaves. An acidic polysaccharide fraction isolated from these unutilized leaves, AL-I, was in our previous work shown to contain pectic polysaccharides. This study aimed to investigate the protective effect of AL-I on ulcerative colitis for the possible application of A. carmichaelii leaves in the treatment of intestinal inflammatory diseases. AL-I was found to alleviate symptoms and colonic pathological injury in colitis mice, and ameliorate the levels of inflammatory indices in serum and colon. The production of short- and branched-chain fatty acids was also restored by AL-I. The observed protective effect could be due to the inhibition of NOD1 and TLR4 activation, the promotion of gene transcription of tight-junction proteins, and the modulation of gut microbiota composition like Bacteroides, Dubosiella, Alistipes and Prevotella,. A regulation of serum metabolomic profiles being relevant to the bacterial change, such as D-mannose 6-phosphate, D-erythrose 4-phosphate and uric acid, was also observed.
Polysaccharides have been assessed as a potential natural active component in Chinese herbal medicine with anti-inflammatory properties. However, the complex and indefinite structures of polysaccharides limit their applications. This study explains the structures and anti-inflammatory potentials of three neutral polysaccharides, RIP-A1 (Mw 1.8 × 104 Da), RIP-B1 (Mw 7.4 × 104 Da) and RIP-B2 (Mw 9.3 × 104 Da), which were isolated from the roots of Isatis indigotica Fort. with sequenced ultrafiltration membrane columns, DEAE-52 and Sephadex G-100. The planar structures and microstructures of RIP-A1, RIP-B1 and RIP-B2 were further determined by HPGPC, GC–MS, methylation analysis, FT-IR, SEM and AFM, in which the structure of RIP-A1 was elucidated in detail using 1D/2D NMR. The Raw 264.7 cells were used for the anti-inflammatory activity in vitro. The results showed that RIP-A1, RIP-B1 and RIP-B2 are all neutral polysaccharides, with RIP-A1 having the smallest Mw and the simplest monosaccharide composition of the three. RIP-A1 is mainly composed of Ara and Gal, except for a small quantity of Rha. Its main structure is covered with glycosidic linkages of T-α-Araf, 1,2-α-Rhap, 1,5-α-Araf, T-β-Galp, 1,2,4-α-Rhap, 1,3,5-α-Araf and 1,6-β-Galp with 0.33:0.12:1.02:0.09:0.45:11.41:10.23. RIP-A1 significantly inhibited pro-inflammatory cytokines (NO, TNF-α, IL-6 and IL-1β) and increased anti-inflammatory cytokines (IL-4) in LPS-stimulated RAW 264.7 cells. Moreover, RIP-A1 could significantly inhibit the mRNA expression of TNF-α, IL-6 and L-1β. It could also activate IKK, p65 and IκBα (the components of the NF-κB signaling pathway). In conclusion, the above results show the structural characterization and anti-inflammatory potentials of RIP-A1 as an effective natural anti-inflammatory drug.
Background
Aconitum carmichaelii Debx. has been widely used as a traditional medicinal herb for a long history in China. It is highly susceptible to various dangerous diseases during the cultivation process. Downy mildew is the most serious leaf disease of A. carmichaelii, affecting plant growth and ultimately leading to a reduction in yield. To better understand the response mechanism of A. carmichaelii leaves subjected to downy mildew, the contents of endogenous plant hormones as well as transcriptome sequencing were analyzed at five different infected stages.
Results
The content of 3-indoleacetic acid, abscisic acid, salicylic acid and jasmonic acid has changed significantly in A. carmichaelii leaves with the development of downy mildew, and related synthetic genes such as 9-cis-epoxycarotenoid dioxygenase and phenylalanine ammonia lyase were also significant for disease responses. The transcriptomic data indicated that the differentially expressed genes were primarily associated with plant hormone signal transduction, plant-pathogen interaction, the mitogen-activated protein kinase signaling pathway in plants, and phenylpropanoid biosynthesis. Many of these genes also showed potential functions for resisting downy mildew. Through weighted gene co-expression network analysis, the hub genes and genes that have high connectivity to them were identified, which could participate in plant immune responses.
Conclusions
In this study, we elucidated the response and potential genes of A. carmichaelii to downy mildew, and observed the changes of endogenous hormones content at different infection stages, so as to contribute to the further screening and identification of genes involved in the defense of downy mildew.
