Crude Polysaccharide Fraction Research Articles

Water-soluble polysaccharides from Piper regnellii (Pariparoba) leaves: Structural characterization and antinociceptive and anti-inflammatory activities

Piper regnellii is a plant popularly known as “Pariparoba” and it is widely used in folk medicine to treat pain, inflammation, among others. This work presents the extraction, purification and characterization of polysaccharides present in the plant leaves and evaluation of their anti-inflammatory and antinociceptive activities. From the crude aqueous extract of P. regnellii leaves, a polysaccharide fraction named PR30R, predominantly constituted of arabinose, galactose and galacturonic acid monosaccharide units, was obtained. Methylation and NMR analysis showed that the main polysaccharides of PR30R are a type II arabinogalactan, formed by a β-D-Galp-(1 → 3) main chain, substituted at O-6 by side chains of β-D-Galp-(1 → 6), which are substituted at O-3 by non-reducing α-L-Araf ends, and a homogalacturonan, formed by →4)-α-D-GalpA-(1→ units. Intraperitoneal administration of the crude polysaccharide fraction PRSF reduced significantly nociception induced by acetic acid in mice at the doses tested, and the PR30R fraction, derived from PRSF, presented antinociceptive and anti-inflammatory effects at a dose of 0.1096 mg/kg (PRSF ED50). These data support the use of the plant leaves in folk medicine as an herbal tea to treat pain and inflammation.

Crude Polysaccharides from Wild-Growing Armillaria mellea—Chemical Composition and Antidiabetic, Anti-Inflammatory, Antioxidant, and Antiproliferative Potential

Armillaria mellea is a commonly harvested and consumed mushroom in Poland. Several activities of polysaccharides from this species have already been reported. However, polysaccharides from A. mellea growing in the wild in Poland have not yet been investigated. This study was conducted to obtain a crude polysaccharide fraction (AmPS) and investigate its chemical composition and biological properties. Our research suggests that A. mellea is a valuable source of polysaccharides, including β-glucans. 1H NMR and high-performance capillary electrophoresis analysis of AmPS revealed the occurrence of polysaccharides consisted of myo-inositol, mannitol, fucose, galactose, glucose, and mannose. Proteins, uronic acids, and phenolics were present only in trace amounts in the AmPS. The AmPS fraction was found to exhibit strong α-glucosidase inhibitory activity; however, no activity in relation to α-amylase was detected. The fraction was also able to inhibit pro-inflammatory enzymes, i.e., lipoxygenase, cyclooxygenase, and hyaluronidase. Moderate antioxidant and antiradical activity of AmPS was revealed by the ORAC and ABTS assay, respectively. The AmPS fraction was found to have antiproliferative potential against gastric and colorectal cancer cell lines (AGS, HT-29). These findings suggest that AmPS may be considered a health-beneficial food ingredient. Since A. mellea is a commonly known edible mushroom with a long tradition of use, its polysaccharides may be considered safe components for direct consumption or production of nutraceuticals.

Crude Polysaccharide Fraction from Rosa rugosa Thunb. Root—Chemical Characterisation, Enzyme Inhibitory, Antioxidant and Antiproliferative Activity

Rosa rugosa Thunb. (Rosaceae) plantations can provide industrial amounts of roots, which contain many biologically active metabolites. Here, we report the first studies of the chemical composition and pharmacological potential of a crude polysaccharide fraction obtained from R. rugosa roots (CPL-Rx). It was found that the roots contained water-soluble and -insoluble sugars, including a large amount of β-glucans (12.95 ± 0.30 g/100 g). The water-soluble fraction was found to be mostly composed of sugars (28.94 ± 0.01%), uronic acids (2.61 ± 0.41%), and polyphenols (4.55 ± 0.12%). High-performance capillary electrophoresis analysis showed that glucose was the prevalent monosaccharide (64.31%), followed by a smaller amount of galactose (11.77%), fructose (11.36%), and arabinose (4.27%). SDS-PAGE followed by liquid chromatography mass spectrometry revealed the presence of unbound protein identified as protein plastid transcriptionally active 14-Like. CPL-Rx was found to inhibit the activity of pro-inflammatory enzymes (cyclooxygenase and hyaluronidase) and to have antioxidant potential in the Trolox equivalent antioxidant capacity assay (0.302 ± 0.01 mmol Trolox/g), DPPH radical scavenging effect (EC50 3.92 ± 0.12 mg/mg DPPH•) and oxygen radical absorbance capacity assay (0.134 ± 0.10 mmol Trolox/g). It also exerted a slight antiproliferative effect on SW480 (colon) and A549 (lung) cancer cell lines. Our research provides the first insights into the composition and pharmacological application of crude polysaccharides from rugosa rose roots. It suggests that CPL-Rx may potentially be used for cosmetic and bio-medical purposes, especially as an antihyaluronidase or chemopreventive agent.

Immune-enhancing effects of polysaccharide extract of by-products of Korean liquor fermented by Saccharomyces cerevisiae

To increase the value of yeast-fermented Korean liquor by-products, we obtained crude polysaccharide (CPS) fractions via ultrasound-assisted extraction and stepwise-gradient ethanol precipitation and investigated their functionality. Nitric oxide production in RAW 264.7 cells was increased following treatment with the CPSs derived from extract. Analysis of the monosaccharide and amino acid composition of the CPS fractions using HPLC revealed that the polysaccharides were mainly composed of glucose (57.2%), mannose (22.6%), and galactose (17.6%), and no amino acids were detected. In addition, a higher concentration of ethanol solvent for fractionation yielded polysaccharides with lower molecular weights (<15 kDa). CPS 3 and 4 fractions increased the production of TNF-α (15 and 17-fold, respectively) and IL-6 (20 and 18-fold, respectively) and iNOS (65 and 35-fold, respectively) expression at concentration 12.5 μg/mL compared with levels in non-treated RAW 264.7 cells. Especially, CPS 4 at 200 and 400 μg/mL significantly increased the proliferation of mouse spleen cells by 126% and 153%, respectively. These results indicated that CPS 4 enhanced the proliferation of mouse spleen cells in vivo, indicating its immune-enhancing effects. Therefore, this research can contribute to the development of eco-friendly extraction techniques and immune-enhancing materials.

A new polysaccharide isolated from Dendrobium offcinale, stimulates aquaporin-3 expression in human keratinocytes

Polysaccharides are the major components of the water extract from Dendrobium officinale, which have been thoroughly studied and found to have favorable moisturizing properties on mammalian skin. However, the structures of polysaccharides on skin moisturizing properties remains unknown. The aim of this study was to extract and characterize D. officinale polysaccharides, which have anti-oxidant activity and the role of promoting effects of aquaporin-3 (AQP3). Two crude polysaccharide fractions were extracted using water and biological fermentation, respectively. The crude polysaccharides were further purified by graded alcohol precipitation, and finally purified polysaccharides DOP-S and DOP-F were obtained by Sephacryl S-300 column chromatography. The average molecular weights of DOP-S and DOP-F were 3.604 × 105 and 1.027 × 105 Da, respectively. Monosaccharide units and structures of DOP-S and DOP-F were further elucidated by gas chromatography-mass spectrometry (GC-MS) and Fourier Transform infrared spectroscopy (FTIR). At the same time, DOP-S and DOP-F induced aquaporin-3 (AQP3) expression was evaluated by immuno-fluorescence analysis. The antioxidant activities of the polysaccharides were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ethylbenzothiazoline-6-sulfonic acid (ABTS) radical scavenging capacities, and ferric reducing antioxidant power (FRAP). The results showed that biological fermentation effectively achieved the hydrolysis of the polysaccharides and stimulated antioxidant activity and AQP3 expression.

Exploration of bioactive prospects of a polysaccharide fraction from Termitomyces heimii against colorectal cancer and broad spectrum bacteria

Termitomyces heimii is a highly cherished wild edible mushroom in India for its pleasant delicacy, rich proximate compositions and profound ethno medicinal applications. A water soluble crude polysaccharide fraction (THP-I) was isolated from dried fruit bodies of T. heimii and purified by GPC. The 1H- NMR spectrum of THP-I exhibited anomeric protons within chemical shifts (δH) between 3.40 to 4.0, indicating that the compound possess –CH and –CH2 functional groups, which resonate with the oxygen atoms of the –OH group, confirmed it as a polysaccharide. THP-I exhibited the MIC values of 62.5 and 125 <mu>g/ml against Staphylococcus aureus and Shigella flexneri respectively. DPPH and FRAP assays showed effective radical scavenging potential by THP-I. In vitro cytotoxicity of THP-I was evaluated by MTT assay and showed notable cellular toxicity against HCT cells at a dose concentration 200 <mu>g/ml. It is first time reported herein that, THP-I has effectively diminished hyperplasia in 1, 2-dimethylhydrazine (DMH) induced colon cancer of Swiss albino rats along with sprouting of goblet cells. Through the present study it is evident that polysaccharide from T. heimii exhibited manifold bioactivities and therefore could be exploited further for its undiscovered potential applications.

Selective activation of the estrogen receptor-β by the polysaccharide from Cynanchum wilfordii alleviates menopausal syndrome in ovariectomized mice

The menopausal syndrome caused by rapid changes in hormone levels greatly influences the quality of life of women. Though hormone replacement therapy (HRT) is widely used to treat the menopausal syndrome, it exhibits many side effects, including the risk of thrombosis, cardiovascular diseases, and increased incidence of breast cancer; thus, diversifying the interest for phytotherapy-based materials as alternatives to HRT. Here, we isolated a crude polysaccharide fraction (CWPF) from Cynanchum wilfordii root that alleviated the ovariectomy-induced uterine atrophy and bone loss without changes in plasma estradiol concentration in mice. Increased plasma levels of follicle-stimulating hormone (FSH), alkaline phosphatase (ALP), osteocalcin (OC) in ovariectomized mice were also reduced to normal levels by CWPF administration. We found that the inhibitory effects of CWPF on menopausal symptoms were mediated by the estrogen receptor β (ER-β) specific activation, not ER-α. Moreover, CWPF treatment suppressed the phosphorylation of Akt, suggesting that CWPF alleviates post-menopausal symptoms by regulating ER-β related Akt signaling pathway. These results demonstrate that the polysaccharides corresponding to CWPF among the water-soluble extracts of CW could be used as a beneficial herbal alternative for the development of therapeutic agents to prevent menopausal syndrome in women.

Physicochemical properties and bioactivities of Lentinula edodes polysaccharides at different development stages

Lentinula edodes polysaccharides from at four different development stages (referred to L1, L2, L3 and L4, respectively) were extracted by hot water method, and graded ethanol precipitation to final concentration of 20%, 50% and 70%, then12 crude polysaccharide fractions (referred to L1P20, L2P20, L3P20; L4P20, L1P50, L2P50, L3P50, L4P50 and L1P70, L2P70, L3P70, L4P70, respectively) were obtained. Physicochemical properties and exoteric bioactivities of the crude polysaccharide fractions were measured. The results of physicochemical properties revealed that extraction yields of P20 fractions were significantly higher than those of P50 and P70 fractions, and the contents of polysaccharide and β-glucan in L3P50 fractions were higher, and the viscosity-average molecular weight reached a maximum at L2, and high molecular weight polysaccharides could be obtained at a low alcohol concentration in P20 fractions, and the glycosidic bonds were found to exist in all crude polysaccharide fractions. These crude polysaccharide fractions showed different bioactivities, wherein the polysaccharides of higher molecular weight in P20 fractions had greater bioactivity. These results showed that immature stage of Lentinula edodes was the optimal harvest time for obtaining higher bioactivity of crude polysaccharides.

A Comparison of Characterization and Its Actions on Immunocompetent Cells of Polysaccharides from Sijunzi Decoction

Sijunzi decoction (SJZD) is a classic recipe in Traditional Chinese Medicine (TCM) with strong immune-enhancement activity. To further understand the characterization and immunomodulatory effect of polysaccharides from SJZD, the monosaccharide compositions of crude polysaccharide (SJZDP), polysaccharide fraction (S-3), and homogeneous polysaccharide (S-3-AG) from SJZD were compared by GC analysis, as well as their immunomodulatory effects on Peyer's patch cells, splenocytes, and macrophages which are related to intestinal immunity, specific immunity, and nonspecific immunity. The results showed that S-3-AG mainly contained Ara with a proportion of 38.9%, while Glc accounted for the largest proportion in S-3 (55.6%) and SJZDP (87.6%). The SJZDP, S-3, and S-3-AG all showed strong capability to stimulate Peyer's patch cells to proliferate and produce IgA and promoted the proliferation and IFN-γ production of splenocytes and increased the NO production and TNF-α production of macrophages. However, S-3 and S-3-AG were able to stimulate splenocytes to secret IL-4, SJZDP had no effect on IL-4 production of splenocytes in the tested concentrations. In addition, S-3 could stimulate the phagocytic activity of macrophages, and S-3-AG restrained the proliferation of macrophages at the concentration of 50–200 µg/mL. These results suggested that SJZDP, S-3, and S-3-AG might have different immunomodulatory effects on intestinal immunity, specific immunity, and nonspecific immunity due to their different monosaccharide compositions. It will provide references for the material basis and mechanism of SJZD immunomodulation activity.

Mycelial polysaccharides of Lentinus edodes (shiitake mushroom) in submerged culture exert immunoenhancing effect on macrophage cells via MAPK pathway.

A crude polysaccharide fraction (termed cLEP) and two derived fractions (termed LEP1 and LEP2) from Lentinus edodes mycelia were purified and characterized. LEP1/-2 were classified as α-type heteropolysaccharides with 1 → 2, 1 → 3, 1 → 4, 1 → 6 linkages. Their monosaccharide components were respectively Gal, Glc, Man, Ara, Fuc, and Rha (molar ratio 10.17:9.75:9.01:1.61:1.26:1), and Glc, Man, Fuc, Rha, and Gal (molar ratio 5.18:4.69:2.85:1.43:1). In vitro culture experiments with macrophage RAW264.7 cells showed no cytotoxic effects of the polysaccharides. Phagocytosis (neutral red uptake) was significantly enhanced by LEP1/-2. Levels of NO, TNF-α and IL-6 were higher in LEP1/-2-treated groups than in cLEP-treated group. qRT-PCR analysis showed that LEP1/-2 had greater enhancing effect on mRNA transcription of iNOS, TNF-α, and IL-6 genes. Western blotting analysis revealed that LEP1/-2 strongly promoted phosphorylation of kinases ERK and JNK, and suggested that they exert immunoenhancing effects via MAPK signaling pathway.

Antimelanogenic effect and whitening of crude polysaccharide fraction extracted from Perilla frutescens Britton var. acuta Kudo

본 연구에서 자소엽 조다당 추출물의 미백 활성을 평가하기 위하여 tyrosinase 억제활성과 melanocyte 세포에서 멜라닌 함량 및 멜라닌 관련 단백질 발현 억제능을 수행하였다. 자소엽 조다당 추출물은 tyrosinase 억제능을 관찰한 결과 자소엽 조다당 추출물의 농도가 증가할수록 활성이 유의적으로 증가하는 것으로 나타...

Comparison of the Biological Activity of Crude Polysaccharide Fractions Obtained from Cedrela sinensis Using Different Extraction Methods

Comparison of the Biological Activity of Crude Polysaccharide Fractions Obtained from Cedrela sinensis Using Different Extraction Methods

Characterization of cell wall polysaccharides from Sicana odorifera fruit and structural analysis of a galactan-rich fraction pectins as side chains

Sicana odorifera is a Brazilian native fruit. In this work, cell wall polysaccharides from S. odorifera pulp were isolated by sequential extraction with water, citric acid, and sodium hydroxide solutions. The monosaccharide composition of crude polysaccharide fractions was determined. The aqueous fractions displayed the highest yields and they were constituted by pectins, having mainly galactans as side chains. The citric acid fraction (SCA) had galactose as the main component. The hemicellulosic fractions consisted mainly of xylose, mannose, galactose, and glucose, suggesting the presence of xyloglucans, xylans and mannans. The SCA fraction was further purified, resulting in a linear galactan (SCAI2). NMR and methylation analysis showed that SCAI2 was a β-(1→4) d-galactan with molar mass of 17,560 g/mol, determined by light scattering. The presence of a linear galactan in free form in fruits is unusual because these polymers usually occur as side chains of type I rhamnogalacturonans.

Antioxidant and neuroprotective effects of crude polysaccharide fractions from Cudrania tricuspidata fruits

Antioxidant and neuroprotective effects of crude polysaccharide fractions from Cudrania tricuspidata fruits

Immunomodulating Activity of Crude Polysaccharide from Inonotus obliquus Sclerotia by Fractionation including MeOH Reflux

To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysac- charide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer's patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production (<TEX>$10{\mu}g/mL$</TEX>) in Peyer's patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.

Neuroprotection against hydrogen peroxide-induced toxicity by Dictyophora echinovolvata polysaccharide via inhibiting the mitochondria-dependent apoptotic pathway.

Neuronal apoptosis caused by toxic stimuli such as oxidative stress is believed to be one of the major reasons in the pathologenesis of neurodegenerative diseases. In the current study, the neuroprotective effects of the crude polysaccharide fraction of edible Dictyophora echinovolvata (DEVP) against H2O2-induced cytotoxicity were investigated using PC12 cells. Following exposure of PC12 cells to 750μM H2O2, a significant reduction in cell viability and the number of FDA-stained viable neurons as well as an increase in the number of PI-stained dead cells were observed. Furthermore, H2O2 treatment significantly upregulated the protein expression of Bax, cleaved caspases 3 and cytosolic cytochrome c, and down-regulated Bcl-2 levels. 2h pre-treatment with VP reversed these changes caused by H2O2, including inhibiting neuronal loss and decreasing Bax, cleaved caspases 3 and cytosolic cytochrome c levels, as well as increasing Bcl-2 levels. These results taken together demonstrated that DEVP provided a substantial neuroprotection against H2O2-induced toxicity in PC12 cells, at least partly through inhibiting the mitochondrial apoptotic pathway. These findings suggested that DEVP might be a potential candidate for further preclinical study for preventing neurodegenerative diseases in which oxidative stress and apoptosis are involved.

Tissue Inhibitor of Metalloproteinase-1 (TIMP-1) and IL-23 Induced by Polysaccharide of the Black Hoof Medicinal Mushroom, Phellinus linteus (Agaricomycetes).

Matrix metalloproteinase-9 (MMP-9) has diverse roles associated with cell growth, migration, invasion, and angiogenesis. Tissue inhibitor of metalloproteinase-1 (TIMP-1) is known to inhibit MMP-9 by complexing with it at a 1:1 ratio. Suppressing MMP-9 activity through the overexpression of TIMP-1 allows for regulation of tumor growth and metastasis by blocking invasion and angiogenesis in the tumor microenvironment. We found that TIMP-1 and interleukin (IL)-23 are induced in RAW264.7 macrophage cells, a cell line established by Abelson leukemia virus transformation from the BALB/c mouse strain, in a dose-dependent pattern, at the transcriptional level by treatment with a crude polysaccharide fraction of Phellinus linteus (CPP) at a range of 10 to 1000 μg/mL. We purified CPP into 2 polysaccharide fractions, Fr-I and Fr-II, and one protein fraction, Fr-III. Among the 3 fractions, Fr-II increased TIMP-1 expression 6.8-fold compared with the control, according to quantitative reverse-transcription polymerase chain reaction (qRT-PCR) analysis in the RAW264.7 culture system. On the other hand, all 3 fractions increased IL-23 expression, with the highest increase brought about by Fr-II. qRT-PCR analysis showed that Fr-I and Fr-II increased IL-17 expression in RAW264.7 cells by 13.3-fold and 19.6-fold, respectively. IL-17 expression in lung tissue was increased 2.1-fold compared with the control group, whereas that in liver tissue was unaltered by oral administration of CPP for 7 days. In a mouse model, qRT-PCR analysis showed that CPP induced liver TIMP-1 and lung IL-17 expressions 8.9-fold and 2.1-fold, respectively, without affecting MMP-9 expression. Our in vitro and in vivo data suggest that inducing TIMP-1 without altering MMP-9 expression by administering the polysaccharide fraction of Ph. linteus could be a novel antitumor or antimetastasis mechanism of polysaccharide from the medicinal mushroom Ph. linteus.

Gastroprotective effect and chemical characterization of a polysaccharide fraction from leaves of Croton cajucara

Croton cajucara Benth. is a tree from the Amazon Forest, where it is known as sacaca. Its leaves and barks are used in medicinal preparations to treat different diseases, including gastric ulcers. The crude polysaccharide fraction (CCP), obtained from the hot aqueous extract of C. cajucara leaves, was able to promote gastroprotection on an ethanol induced gastric ulcer model. Therefore, a bioguided fractionation was performed to isolate the active polysaccharide fraction. After freezing-thawing, ultrafiltration and dialyses at 100, 50, and 25kDa cut-off membranes, fraction 25R was obtained. It contained glucose, galactose, rhamnose, arabinose, galacturonic acid and mannose in a 7:5:5:3:1:1 molar ratio approximately, and had a Mw of 42,840g/mol. Methylation analysis and NMR spectroscopy indicated that 25R is a very complex polysaccharide fraction containing type I rhamnogalacturonan, arabinan, type I arabinogalactan, type II arabinogalactan, rhamnan, starch and mannan. It was able to reduce ethanol-induced gastric ulcers in rats, through preservation of mucus and GSH levels.

Polysaccharides from Arctium lappa L.: Chemical structure and biological activity

The plant Arctium lappa L. is popularly used to relieve symptoms of inflammatory disorders. A crude polysaccharide fraction (SAA) resulting of aqueous extraction of A. lappa leaves showed a dose dependent anti-edematogenic activity on carrageenan-induced paw edema, which persisted for up to 48h. Sequential fractionation by ultrafiltration at 50kDa and 30kDa cut-off membranes yielded three fractions, namely RF50, RF30, and EF30. All these maintained the anti-edematogenic effect, but RF30 showed a more potent action, inhibiting 57% of the paw edema at a dose of 4.9mg/kg. The polysaccharide RF30 contained galacturonic acid, galactose, arabinose, rhamnose, glucose, and mannose in a 7:4:2:1:2:1 ratio and had a Mw of 91,000g/mol. Methylation analysis and NMR spectroscopy indicated that RF30 is mainly constituted by a type I rhamnogalacturonan branched by side chains of types I and II arabinogalactans, and arabinan.

Structure Characterization and Immunomodulating Effects of Polysaccharides Isolated from Dendrobium officinale.

A crude polysaccharide fraction (cDOP) has been determined to be the characteristic marker of Dendrobium officinale, an expensive tea material in Asia, but its chemistry and bioactivity have not been studied. In work reported here, cDOP was destarched (DOP, 90% yield) and separated into two subfraction polysaccharides, DOPa and DOPb, which were characterized by monosaccharide composition and methylation analyses and spectral analyses (FT-IR and (1)H and (13)C NMR). Both are composed of mannose and glucose at similar ratios and have a similar structure with a backbone of 1,4-linked β-D-mannopyranosyl and β-D-glucopyranosyl residues. Significant differences were observed only in their molecular weights. Bioassay using mouse macrophage cell line RAW264.7 indicated that DOP and its two subfractions enhance cell proliferation, TNF-α secretion, and phagocytosis in a dose-dependent manner. They also induced the proliferation of lymphocytes alone and with mitogens. DOPa and DOPb are thus proven to be major, active polysaccharide markers of D. officinale.