Athyrium Multidentatum Research Articles

Protective effect of phosphorylated Athyrium multidentatum (Doll.) Ching polysaccharide on vascular endothelial cells in vitro and in vivo.

The purpose of this study was to prepare phosphorylated Athyrium multidentatum (Doll.) Ching polysaccharide (PPS) and investigate its protective effect on vascular endothelial cells (VECs) in vitro and in vivo and the underlying mechanisms. Sodium tripolyphosphate (STPP) and sodium trimetaphosphate (STMP) were used as phosphorylation reagents and PPS was characterized by Fourier transform infrared (FT-IR), 13 C nuclear magnetic resonance (13 C NMR) and 31 P nuclear magnetic resonance (31 P NMR) spectra. Chemical analysis demonstrated that PPS was composed of mannose, glucosamine, rhamnose, glucuronic acid, galacturonic acid, galactosamine, glucose, galactose, xylose, arabinose, and fucose with a molar ratio of 11.36:0.42:4.03:1.12:1.81:0.26:33.25:24.12:6.85:14.46:2.32 and a molecular weight of 28,837 Da. Results from in vitro and in vivo assays revealed that PPS protected human umbilical vein endothelial cells (HUVECs) against H2 O2 -induced oxidative injury and attenuated D-galactose-induced VECs damage in mice. RNA sequencing (RNA-seq) analysis identified 18 differentially expressed genes (DEGs) between D-galactose-treated and PPS-pretreated mice abdominal aorta. A deep analysis of these DEGs disclosed that PPS regulated the expression of genes involved in the functions of vascular endothelium repairment, cell growth and proliferation, cell survival and apoptosis, inflammation, angiogenesis and antioxidant, indicating that these biological processes might play crucial roles in the protective actions of PPS on VECs.

Effect of selenized Athyrium multidentatum (Doll.) Ching polysaccharides on TNF / MAPKs / NF‐κB signaling pathways in RAW 264.7 cells

Polysaccharides from Athyrium multidentatum (Doll.) Ching (AMC) were fractionated and selenylated by HNO3-Na2SeO3 method. Four selenized polysaccharides were synthesized and fully characterized by ICP-MS, UV, FT-IR, 1H NMR, high-performance gel permeation chromatography, high-performance liquid chromatography (HPLC), and SEM. Chemical analysis demonstrated that the selenized polysaccharides were composed of mannose (Man), rhamnose (Rha), glucuronic acid (Glc A), glucose (Glc), galactose (Gal), arabinose (Ara), and fucose (Fuc) with different molar ratios and molecular weights. The immunoactivity results showed that the selenized polysaccharides elevated cell proliferation, phagocytic activity, and promoted the production of TNF-α, IL-6, IL-1β, IFN-γ, IL-10, NO, and reactive oxygen species in RAW 264.7 cells, indicating marked immune enhancement activities. RT-qPCR and Western blot assays revealed that the selenized AMC polysaccharides upregulated the expression levels of many key factors involved in TNF, MAPKs, and NF-κB signaling pathways. TNF/MAPKs/NF-κB signaling pathways might play crucial roles in the immune function of selenized AMC polysaccharides. Practical applications Polysaccharide is an essential nutrient in Athyrium multidentatum (Doll.) Ching. The selenized AMC polysaccharide possesses the activities of organic selenium and polysaccharide, making it abroad application prospects in the fields of medicine, food, and cosmetics.

Structural characteristics and immune-enhancing activity of fractionated polysaccharides from Athyrium Multidentatum (Doll.) Ching.

Polysaccharides coded as CP were extracted from Athyrium Multidentatum (Doll.) Ching and then fractionated into five fractions (FP-1, FP-2, FP-3, FP-4 and FP-5). A purified polysaccharide designated as FP-3-4 was prepared from FP-3 by Sephadex G-100 column chromatography. Chemical analysis disclosed that CP and these fractions were heteropolysaccharides and mainly composed of glucose, galactose, arabinose, mannose, rhamnose, xylose, fucose, ribose and uronic acid with different molar ratios. They presented different images of SEM. FP-3-4 was highly branched polymers with sixteen types of linkages. The in vitro immunomodulatory results stated that CP and these fractions could promote macrophage proliferation, enhance macrophage phagocytosis and increase the production of NO, TNF-α, IFN-γ, IL-1β, IL-6, IL-10 and IL-2, indicating remarkable immune enhancement activities. RNA sequencing analysis revealed that CP and FP-3 induced macrophage activation mainly through MAPK and alternative NF-κΒ signaling pathways via CD14/TLR4 and Dectin-2 receptors, which were verified by RT-qPCR and western blot.

Chemical characteristics and cytoprotective activities of polysaccharide fractions from Athyrium Multidentatum (Doll.) Ching

Five polysaccharide fractions (PS-1, PS-2, PS-3, PS-4 and PS-5) were successfully isolated from Athyrium Multidentatum (Doll.) Ching by anion-exchange column chromatography. Their in vitro cytoprotective activities and the underlying mechanisms were explored in this paper. Chemical analysis suggested that the five polysaccharide fractions were heteropolysaccharides with different molecular weights and monosaccharide compositions. Treatment with these polysaccharide fractions could increase cell viabilities, superoxide dismutase/catalase activities, nitric oxide contents, mitochondrial membrane potential levels and Bcl-2/Bax ratios, and reduce cell apoptosis, intracellular reactive oxygen species production and malondialdehyde contents in H2O2-damaged cells. Moreover, these polysaccharide fractions enhanced the mRNA expression levels of PI3K, Akt, FOXO3a, Nrf2 and HO-1 and PS-4 exhibited the most powerful effects on the mRNA expression of these genes. Current findings suggested that the polysaccharide fractions decreased H2O2-induced apoptosis of HUVECs. The activation of PI3K/Akt/FOXO3a and Nrf2/HO-1 signaling pathways might be involved in the protective mechanisms of the active fractions. The polysaccharides might be one of the key bioactive ingredients of Athyrium Multidentatum (Doll.) Ching for the treatment of oxidative damage.

Response of total phenols, flavonoids, minerals, and amino acids of four edible fern species to four shading treatments.

Total phenols, flavonoids, minerals and amino acids content were investigated in leaves of four fern species grown under four shading treatments with different sunlight transmittance in 35% full sunlight (FS), 13% FS, 8% FS and 4% FS. The leaves of four fern species contain high levels of total phenols and flavonoids, abundant minerals and amino acids, and these all were strongly affected by transmittance. Total phenols and flavonoids content were significantly positively correlated with transmittance, while minerals and total amino acids content were significantly negatively correlated with transmittance, a finding that supports research into how higher light intensity can stimulate the synthesis of phenols and flavonoids, and proper shading can stimulate the accumulation of minerals and amino acids. Matteuccia struthiopteris (L.) Todaro (MS) had the highest total phenols content, Athyrium multidentatum (Doll.) Ching (AM) showed the highest total amino acids, total essential amino acids content, Osmunda cinnamomea (L) var. asiatica Fernald (OCA) exhibited the highest total non-essential amino acids and flavonoids content. Pteridium aquilinum (L.) Kuhn var. latiusculum (Desy.) Underw. ex Heller (PAL) exhibited the highest minerals content. This research can provide a scientific basis for the cultivation and management of those four fern species.

Structural Characterization and Antioxidant Activity of Polysaccharides from Athyrium multidentatum (Doll.) Ching in d-Galactose-Induced Aging Mice via PI3K/AKT Pathway.

The purpose of this study was to characterize the polysaccharides from Athyrium multidentatum (Doll.) Ching (AMC) rhizome and explore the protective mechanism against d-galactose-induced oxidative stress in aging mice. Methods: A series of experiments, including molecular weight, monosaccharide composition, Fourier transform infrared (FT-IR) spectroscopy, and 1H nuclear magnetic resonance (1H NMR) spectroscopy were carried out to characterize AMC polysaccharides. The mechanism was investigated exploring d-galactose-induced aging mouse model. Quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) and western blotting assays were performed to assess the gene and protein expression in liver. Key findings: Our results showed that AMC polysaccharides were mainly composed of mannose (Man), rhamnose (Rha), glucuronic acid (Glc A), glucose (Glc), galactose (Gal), arabinose (Ara), and fucose (Fuc) in a molar ratio of 0.077:0.088:0.09:1:0.375:0.354:0.04 with a molecular weight of 33203 Da (Mw). AMC polysaccharides strikingly reversed d-galactose-induced changes in mice, including upregulated phosphatidylinositol 3-kinase (PI3K), Akt, nuclear factor-erythroid 2-related factor 2 (Nrf2), forkhead box O3a (FOXO3a), and hemeoxygenase-1 (HO-1) mRNA expression, raised Bcl-2/Bax ratio, downregulated caspase-3 mRNA expression, enhanced Akt, phosphorylation of Akt (p-Akt), Nrf2 and HO-1 protein expression, decreased caspase-3, and Bax protein expression. Conclusion: AMC polysaccharides attenuated d-galactose-induced oxidative stress and cell apoptosis by activating the PI3K/AKT pathway, which might in part contributed to their anti-aging activity.

Striatisporolide A, a butenolide metabolite from Athyrium multidentatum (Doll.) Ching, as a potential antibacterial agent.

The present study aimed to investigate the antibacterial activity of striatisporolide A (SA) against Escherichia coli (E. coli) and the underlying mechanism. Antibacterial activity was evaluated according to the inhibitory rate and zone of inhibition. The antibacterial mechanism was investigated by analyzing alkaline phosphatase (AKP) activity and ATP leakage, protein expression, cell morphology and intracellular alterations in E. coli. The results demonstrated that SA exerted bacteriostatic effects on E. coli in vitro. AKP activity and ATP leakage analysis revealed that SA damaged the cell wall and cell membrane of E. coli. SDS-PAGE analysis indicated that SA notably altered the level of 10 and 35 kDa proteins. Scanning electron microscopy and transmission electron microscopy analyses revealed marked alterations in the morphology and ultrastructure of E. coli following treatment with SA. The mechanism underlying the antimicrobial effects of SA against E. coli may be attributed to its actions of disrupting the cell membrane and cell wall and regulation of protein level. The findings of the present study provide novel insight into the antimicrobial activity of SA as a potential natural antibacterial agent.

Growth, Secondary Metabolites and Enzyme Activity Responses of Two Edible Fern Species to Drought Stress and Rehydration in Northeast China

The drought resistance mechanism of Matteuccia struthiopteris (L.) Todar. and Athyrium multidentatum (Doll.) Ching were measured under natural drought exposure. The results showed that the two edible fern species showed stronger resistance in the early stages of drought, mainly expressed as the decrease of relative leaf water content (RLWC), increase of osmotic substances, secondary metabolites such as flavonoids (FC), total phenols (TPC), proantho cyanidins (PCC) content and enzyme activity (superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX)). The higher RLWC, FC, TPC, PCC and abscisic acid (ABA) content and lower H2O2 content indicates the stronger non-enzymatic antioxidant system and drought resistance of A. multidentatum. However, the proline (Pro) content changed slowly, and the synthesis of soluble protein (SP), total phenols, proantho cyanidins and ABA, SOD activity of two fern species were inhibited in the late stages of drought stress. This study can provide a scientific basis for the cultivation and utilization of edible fern species under forest in Northeast China.

Anti-inflammatory action of Athyrium multidentatum extract suppresses the LPS-induced TLR4 signaling pathway

Ethnopharmacological relevanceThe aerial part of Athyrium multidentatum (Doll.) Ching (AM) is widely used in the northeastern region of China as an edible wild herb, but its medicinal value, especially its anti-inflammatory effect, has not been fully explored. Aim of the studyTo investigate the anti-inflammatory activity of AM and clarify the anti-inflammatory mechanism involving the TLR4 signaling pathway using a lipopolysaccharide (LPS)-induced inflammatory model. Materials and methodsAM ethanol extract was used as the experimental material to investigate the effect that the extract has on the production of pro-inflammatory mediators (NO, PGE2, TNF-α, IL-1β and IL-6); changes in LPS-induced peritoneal macrophages (PMs); and TLR4-mediated intracellular events, including MAPKs (ERK, JNK, and p38) and IκB-α in the MyD88-dependant pathway and IRF3, STAT1, and STAT3 in the TRIF-dependent pathway. In in vivo experiments, we established an LPS-induced acute lung injury (ALI) model and investigated the cell count and cytokine (TNF-α, IL-1β and IL-6) levels in bronchoalvelar lavage fluid (BALF) of C57BL6 mice. Histological changes in the lung tissues were observed with H&E staining. ResultsAM extract inhibited NO and PGE2 by suppressing their synthetase (iNOS and COX-2) gene expression in LPS-induced PMs; the secretion of IL-6, IL-1β, and TNF-α also deceased via the down-regulation of mRNA levels. Furthermore, the TLR4-mediated intracellular events involved the phosphorylated forms of MAPKs (ERK, JNK) and IκB-α in the MyD88-dependent pathway and the TRIF-dependent pathway (IRF3, STAT1, STAT3), and the relevant proteins were expressed at low levels in the AM extract groups. In in vivo experiments, the cell count and cytokine (TNF-α, IL-1β and IL-6) levels in BALF decreased significantly in a dose-dependent manner in the AM extract groups. The lung tissue structure exhibited dramatic damage in the LPS group, and the damaged area decreased in the AM extract groups; in particular, the effect of 10 mg/kg extract was similar to that of the positive control dexamethasone (DEX). ConclusionThe findings demonstrate that AM protects against LPS-induced acute lung injury by suppressing TLR4 signaling, provide scientific evidence to support further study of the safety of anti-inflammatory drugs and indicate that AM can be used as an anti-inflammatory and anti-injury agent to prevent pneumonia caused by microbial infection.

Athyrium multidentatum (Doll.) Ching extract induce apoptosis via mitochondrial dysfunction and oxidative stress in HepG2 cells

Athyrium multidentatum (Doll.) Ching (AMC), a unique and nutritious potherb widely distributed in china, has been extensively used in traditional Chinese medicine. Previous studies indicated that AMC extract exhibited antioxidant and antitumor properties. However, the chemical composition of AMC and molecular mechanism of AMC toxicity to HepG2 cells have not yet been elucidated. Hence, this study aimed to investigate the chemical compositions and the underlying mechanisms of the antiproliferative and apoptotic effects of AMC on HepG2. HPLC-MS analysis showed that AMC contain five compounds with chlorogenic acid accounting for 43 percent. Also, AMC strongly inhibited the cell growth and induced apoptosis and cell cycle arrest in HepG2 cells by significantly upregulating the protein expressions of Fas, Fas-L, Bax/Bcl-2, cyto-c, cleaved caspase-3, and PARP in a dose-dependent manner, which indicates AMC induces apoptosis in HepG2 cells through both intrinsic and extrinsic pathways. Moreover, AMC provoked the production of ROS, H2O2, and NO, modulating the PI3K/Akt, MAPK, NFκB and Nrf2 pathways and their downstream transcriptional cascades, ultimately evoked oxidative stress and apoptosis in HpeG2 cells. Further in vivo experiments demonstrated that AMC significantly suppressed the tumor growth, suggesting that AMC may be a novel promising agent for hepatocellular carcinoma treatment.

Cytoproliferative and Cytoprotective Effects of Striatisporolide A Isolated from Rhizomes of Athyrium multidentatum (Doell.) Ching on Human Umbilical Vein Endothelial Cells.

Objectives: The aim of this study was to investigate the proliferative and protective effects of striatisporolide A (SA) obtained from the rhizomes of Athyrium multidentatum (Doell.) Ching on human umbilical vein endothelial cells (HUVECs). Methods: Cell viability was measured by the MTT method. Cell apoptosis was determined by flow cytometry. Intracellular ROS was measured by the 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe. Results: The viability rate in cells treated with 100 µM SA alone was increased to 128.72% ± 0.19% and showed a significant difference compared with the control group (p < 0.05). Meanwhile, SA augmented the cell viabilities in H2O2-treated HUVECs, and the cell viability was enhanced to 56.94% ± 0.13% (p < 0.01) when pre-incubated with 50 µM SA. The cell apoptosis rates were reduced to 2.17% ± 0.20% (p < 0.05) and 3.1% ± 0.34% (p < 0.01), respectively, after treatment with SA alone or SA/H2O2. SA inhibited the overproduction of reactive oxygen species (ROS) in HUVECs induced by H2O2 and the fluorescent intensity was abated to 9.47 ± 0.61 after pre-incubated with 100 μM SA. Conclusions: The biological activities of SA were explored for the first time. Our results stated that SA exhibited significant cytoproliferative and minor cytoprotective effects on HUVECs. We presume that the mechanisms of the proliferation and protection actions of SA involve interference with the generation of ROS and the cell apoptosis. These findings provide a new perspective on the biological potential of butenolides.

Content and antioxidant activity of total polyphenols and tannins from Athyrium multidentatum （Doll.）Ching

Objective To determine the content and to evaluate the antioxidant activity of total polyphenols and tannins from Athyrium multidentatum（Doll.）Ching rhizome. Methods Folin phenol reagent-casein method was used to determine the content of total polyphenols and tannins. Spectrophotometry was used to evaluate the antioxidant activity. Results The content of total polyphenols and tannins was 24.61%and 4.59%, respectively. The total polyphenols and tannins exhibited strong reducing power and hydroxyl radicals/superoxide radicals scavenging capability. Conclusion The antioxidant activity of total polyphenols and tannins from Athyrium multidentatum（Doll.）Ching rhizome is significant and further investigation is deserved in the future. Key words: Athyrium multiclentatum （Doll.） Ching; Tannins; Polyphenols; Antioxidants （TCD）Spectrophotometry

STUDY ON DROUGHT STRESS OF SIX COMMON FERNS IN NORTH CHINA

Six common ferns (Pteridium aquilinum var. latiusculum, Adiantum pedatum, Polystichum tripteron, Matteuccia struthiopteris, Adiantum capillus-veneris, Athyrium multidentatum) in north China were selected to test for drought stress. During drought stress, ferns were randomly selected to determine photosynthetic rate, soil moisture content, chlorophyll content, leaf water content, leaf water-retention ability and electric conductivity. The plant growth information was investigated in different water conditions. The results showed that: the ranks of drought resistance of the six ferns were Adiantum capillus-veneris > Adiantum pedatum > Polystichum tripteron > Matteuccia struthiopteris > Athyrium multidentatum > Pteridium aquilinum var. latiusculum. Drought resistance was made clear by soil moisture content, leaf water content and net photosynthetic rate. The results indicated that the soil moisture content, leaf water content and net photosynthetic rate were the most reliable indexes; the leaf relative electric conductivity, chlorophyll concentration could be used as auxiliary indexes; and the leaf moisture retaining power and chlorophyll a/b rate partly reflected the drought tolerance, as reference indexes. Matteuccia struthiopteris applied in urban greening is abundant in parks of Beijing at present. The results of statistical analysis of related data showed that Adiantum capillus-veneris, Adiantum pedatum and Polystichum tripteron have strong drought tolerance, and good visual effect. Therefore there is an extremely widespread application prospect in landscaping of north China, especially in Beijing.

Antioxidant properties of different molecular weight polysaccharides from Athyrium multidentatum (Doll.) Ching

Different molecular weight polysaccharides were prepared by degradation of polysaccharides extracted from Athyrium multidentatum (Doll.) Ching rhizome (CPA) with hydrogen peroxide and ascorbic acid. Four low molecular polysaccharides derivatives (CPA-1, CPA-2, CPA-3 and CPA-4) were successfully obtained and had their antioxidant activities investigated employing various established in vitro systems. All CPA derivatives showed pronounced antioxidant activity, and had stronger antioxidant ability than CPA in certain tests. CPA-1 exhibited the strongest scavenging ability on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical among all samples, and the IC50 value was 25μg/mL. CPA-2 possessed the highest scavenging ability against superoxide radical at 200μg/mL. The scavenging activity of CPA-4 on hydroxyl radical was higher than CPA from 120 to 200μg/mL. The mechanism on influence the antioxidant activity of CPA and its degraded derivatives was indicated.

Antioxidant activity of polysaccharide fractions extracted from Athyrium multidentatum (Doll.) Ching

Crude polysaccharides were extracted from Athyrium multidentatum (Doll.) Ching (AMC) rhizome and fractionated by DEAE-Cellulose 52 ion-exchange column chromatography. Two polysaccharide fractions (F1 and F2) were obtained and had their antioxidant activities investigated employing various established in vitro systems. All fractions possessed considerable antioxidant activity. Chemical analysis suggested that F1 and F2 were neutral heteropolysaccharide in which glucose was the major component. Available data suggested that the molecular weight and sulfate content played very important roles on antioxidant activity. Our results indicated that the polysaccharides may contribute to the medicinal functions of AMC.

Bioactivity-guided fractionation for antioxidant property of Athyrium multidentatum (Doll) Ching. rhizome

In this paper, methanolic extract of Athyrium multidentatum (Doll) Ching. (AMC) rhizome was fractionated into three fractions (petroleum ether fraction (PEF), ethyl acetate fraction (EAF) and n-butanol fraction (BF)) on the basis of polarity. Each fraction had their antioxidant activities estimated employing various established in vitro systems, including superoxide/hydroxyl radical scavenging activity and reducing power. BF showed the strongest antioxidant activity and was chromatographed on a silica gel column followed by gradient elution. Fractions with CH2Cl2 – EtOAc 10:1, CH2Cl2 – EtOAc 5׃1~1׃1 and EtOAc – CH3OH 10׃1~1׃1 as eluent showed significant antioxidant activity against superoxide anion radical and reducing power. All the results showed that AMC could be potential rich sources of natural antioxidants. Key words: Athyrium multidentatum (Doll.) Ching, antioxidant activity, radical scavenging activity, reducing power, active fraction.

Antioxidant activity of polysaccharides extracted from Athyrium multidentatum (Doll) Ching

In this paper, total polysaccharides were respectively prepared from rhizome and aerial part of Athyrium Multidentatum (Doll) Ching. (AMC) and had their antioxidant activities investigated employing various established in vitro systems, including superoxide/hydroxyl radical scavenging activity and reducing power. Sulfate content and molecular weights of polysaccharides from rhizome (PR) and aerial part (PA) were determined. PR and PA exhibited stronger scavenging capacity on superoxide radicals than vitamin C (Vc) at concentrations from 22.22 to 44.44 μg/ml. The sulfate content in PR and PA were 3.45 and 4.68%, respectively. PR and PA consisted of different polysaccharide fractions. Available data obtained with in vitro models suggested that the ratio of sulfate content was an effective indicator to antioxidant activity of the samples. Key words: Antioxidant activity, Athyrium Multidentatum (Doll.) Ching., polysaccharide, radical scavenging activity, reducing power.

Structure changes and succession dynamic of the natural secondary forest after severe fire interference

The structure and dynamic succession law of natural secondary forest after severe fire interference in recent 20 years were studied by adopting the method of deducing time series from the spatial sequence of vegetation in Heihe region, Heilongjiang, China. Two typical and widely distributed forest types in the study area, namely forest type A and forest type B, were selected as study subjects. Forest type A is pure broadleaf forest or broadleaf mixed forest mainly composing of superior Betula platyphylla and Populus davidiana in the area with gradient 25°. Species richness, vegetation coverage, important value, and similarity index of community in different layers (Herb, shrub, small tree, and arbor layers) were investigated and analyzed for the two typical forests. The results show that after fire interference, the species richness and coverage in each layer in forest type A were higher than that in forest type B. Both for forest type A and B, with elapse of post-fire years, the species richness and coverage of herbs and shrubs showed a decline tendency, while those of arbor layer present a rising tendency. Through comparison of the important values of species in each layer and analysis of community structure changes, the dynamic process of post-fire vegetation succession for forest type A and B was separately determined. Post-fire 80 years’ succession tendency of forest type A is B. platyphylla and Larix gmelinii mixed forest. Its shrub layer is mainly composed of Corylus heterophylla and Vaccinium uliginosum, and herb layer is dominated by Carex tristachya, Athyrium multidentatum, and Pyrola incarnate; whereas, the post-fire 80 years’ succession of forest type B is Q. mongolica and B. davurica mixed forest. Its shrub layer is mainly composed of lespedeza bicolar and corylus heterophylla and herb layer is dominated by Carex tristachya, Asparagus densiflorus, and Hemerocallis minor.