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RAW264.7 Cell Activating Glucomannans Extracted from Rhizome of Polygonatum sibiricum
Yelithao, Khamphone,Surayot, Utoomporn,Lee, Ju Hun,You, SangGuan The Korean Society of Food Science and Nutrition 2016 Preventive Nutrition and Food Science Vol.21 No.3
Water-soluble polysaccharides isolated from the rhizome of Polygonatum sibiricum and fractionated using ionexchange chromatography were investigated to determine their structure and immunostimulating activity. Crude and fractions ($F_1$ and $F_2$) consisted of carbohydrates (85.1~88.3%) with proteins (4.51~11.9%) and uronic acid (1.79~7.47%), and included different levels of mannose (62.3~76.3%), glucose (15.2~20.3%), galactose (4.35~15.3%), and arabinose (4.00~7.65%). The crude contained two peaks with molecular weights (Mw) of $151{\times}10^3$ and $31.8{\times}10^3$, but $F_1$ and $F_2$ exhibited one major peak with Mw of $103{\times}10^3$ and $628{\times}10^3$, respectively. Little immunostimulatory activity was observed by the crude; however, $F_1$ and $F_2$ significantly activated RAW264.7 cells to release nitric oxide and various cytokines, suggesting they were potent immunostimulators. The backbone of the most immunostimulating fraction ($F_1$) was ($1{\rightarrow}4$)-manno- and ($1{\rightarrow}4$)-gluco-pyranosyl residues with galactose and glucose attached to O-6 of manno-pyranoside.
RAW264.7 Cell Activating Glucomannans Extracted from Rhizome of Polygonatum sibiricum
Khamphone Yelithao,Utoomporn Surayot,Ju Hun Lee,SangGuan You 한국식품영양과학회 2016 Preventive Nutrition and Food Science Vol.21 No.3
Water-soluble polysaccharides isolated from the rhizome of Polygonatum sibiricum and fractionated using ionexchange chromatography were investigated to determine their structure and immunostimulating activity. Crude and fractions (F₁ and F₂) consisted of carbohydrates (85.1∼88.3%) with proteins (4.51∼11.9%) and uronic acid (1.79∼7.47%), and included different levels of mannose (62.3∼76.3%), glucose (15.2∼20.3%), galactose (4.35∼15.3%), and arabinose (4.00∼7.65%). The crude contained two peaks with molecular weights (Mw) of 151×10³ and 31.8×10³, but F₁ and F₂ exhibited one major peak with Mw of 10³×10³ and 628×10³, respectively. Little immunostimulatory activity was observed by the crude; however, F₁ and F₂ significantly activated RAW264.7 cells to release nitric oxide and various cytokines, suggesting they were potent immunostimulators. The backbone of the most immunostimulating fraction (F₁) was (1→4)- manno- and (1→4)-gluco-pyranosyl residues with galactose and glucose attached to O-6 of manno-pyranoside.
Yelithao, Khamphone,Surayot, Utoomporn,Park, WooJung,Lee, SangMin,Lee, Dae-Hee,You, SangGuan Elsevier 2019 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Vol.122 No.-
<P><B>Abstract</B></P> <P>The aqueous polysaccharide from <I>Polygonatum sibiricum</I> was extracted and fractionated using anion-exchange chromatography to obtain F<SUB>1</SUB> fraction. The F<SUB>1</SUB> was chemically sulfated and partially acid-hydrolyzed for the production of its over-sulfated (OS<SUB>1,2,3</SUB>) and hydrolyzed (HP<SUB>1,2,3</SUB>) derivatives, in which the sulfate content of OS<SUB>1,2,3</SUB> was 7.5–17.1%, and the M<SUB>w</SUB> of HP<SUB>1,2,3</SUB> ranged from 18.2 × 10<SUP>3</SUP> to 57.3 × 10<SUP>3</SUP> g/mol. Considerable RAW264.7 cell activation was observed by HP<SUB>1,2,3</SUB> with NO production of 34.9, 44.3 and 42.7 μM, respectively, as well as the mRNA expression of cytokines (IL-1β, IL-6, IL-10 and IL-12). NK cell cytotoxicity against HT-29 cell was facilitated by OS<SUB>1,2,3</SUB> treatment with the increased gene expressions of INF-γ, Granzyme-B, perforin, NKG2D, and FasL. RAW264.7 cells appeared to be activated via MR and TLR4 mediated signaling pathway, but CR3 and TRL2 might play a main role in stimulating NK cells. Overall, the present study suggests the potential application of polysaccharides from <I>P. sibiricum</I> in functional foods and pharmacological industries.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Polysaccharide was extracted and purified from <I>Polygonatum sibiricum</I>. </LI> <LI> Fraction 1 (F<SUB>1</SUB>) was over-sulfated and acid hydrolyzed. </LI> <LI> The M<SUB>w</SUB> of F<SUB>1</SUB> appeared to influence the RAW264.7 cell activation. </LI> <LI> The sulfation was beneficial for improving its NK cell cytotoxicity. </LI> </UL> </P>
Tabarsa, Mehdi,Dabaghian, Elham Hashem,You, SangGuan,Yelithao, Khamphone,Cao, RongAn,Rezaei, Masoud,Alboofetileh, Mehdi,Bita, Seraj Elsevier 2020 INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES Vol.148 No.-
<P><B>Abstract</B></P> <P>Polysaccharides from <I>Nizamuddinia zanardinii</I> were extracted using water at elevated temperature and fractionated by a DEAE Sepharose FF column yielding four fractions (F<SUB>1</SUB>-F<SUB>4</SUB>). Crude and fractions were composed of neutral sugars (50.8–57.4%), proteins (10.8–18.1%), sulfates (7.5–17.3%) and uronic acids (3.5–7.7%). Various levels of galactose (13.4–44.4%), fucose (34.1–40.1%), mannose (14.1–33.2%) and xylose (7.4–15.2%) formed the building blocks of the polysaccharide structures. The weight average molecular weights (<I>M</I> <SUB>w</SUB>) of polysaccharides varied between 40.3 and 1254.4 × 10<SUP>3</SUP> g/mol. F<SUB>3</SUB> polysaccharide was the most active fraction stimulating RAW264.7 murine macrophage cells to secrete NO, TNF-α, IL-1β and IL-6, and activating NK cells to release TNF-α, INF-γ, granzyme-B, perforin, NKG2D and FasL through NF-κB and MAPKs signaling pathways. Highly-branched F<SUB>3</SUB> polysaccharide mainly consisted of (1 → 2)-Fuc<I>p</I>, (1 → 2,3)-Man<I>p</I>, (1 → 3)-Gal<I>p</I>, (1 → 2)-Man<I>p</I>, (1 → 3)-Man<I>p</I>, (1 → 2,3,4)-Man<I>p</I> and (1 → 2,3,6)-Man<I>p</I> residues with great amount of (→1)-Fuc<I>p</I> and (→1)-Xyl<I>p</I>. Sulfates substituted at C-2 of fucose and galactose residues. Overall, fucoidan from <I>N. zanardinii</I> showed immense potency in boosting immune system through macrophages and NK cells activations and therefore suitable for further exploration in immune-mediated biomedical applications.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Water-soluble fucoidans were isolated from <I>Nizamuddinia zanardinii</I>. </LI> <LI> Fucoidans activated RAW264.7 cells to release proinflammatory mediators. </LI> <LI> Fucoidans stimulated natural killer cells to produce TNF-α, INF-γ and perforin. </LI> <LI> RAW264.7 and NK cells were activated through NF-κB and MAPK signaling pathways. </LI> <LI> Most immunoenhancing polysaccharide contained high (1 → 2)-fucopyranosyl units. </LI> </UL> </P>
Bahramzadeh, Saman,Tabarsa, Mehdi,You, SangGuan,Yelithao, Khamphone,Klochkov, Vladimir,Ilfat, Rakhmatullin Elsevier 2019 Journal of Functional Foods Vol.52 No.-
<P><B>Abstract</B></P> <P>Polysaccharides from <I>Boswellia carterii</I> were isolated using hot water and fractionated on a DEAE Sepharose Fast Flow column to evaluate their structural characteristics and macrophage stimulating capacities. Crude and fractionated polysaccharides (BCF<SUB>1</SUB> and BCF<SUB>2</SUB>) were mainly consisted of different amounts of neutral sugars (63.7–83.3%) and uronic acids (6.2–22.3%). The weight average molecular weights (<I>M</I> <SUB>w</SUB>) of polysaccharides greatly varied between 295.5 and 504.7 × 10<SUP>3</SUP> g/mol. The BCF<SUB>2</SUB> was a highly active polysaccharide on RAW264.7 murine macrophage cells inducing considerable release of nitric oxide and proinflammatory cytokines including TNF-α, IL-1β and IL-6. The presence of p-NF-κB, p-JNK, p-ERK and p-p38 proteins in the cytoplasm of the stimulated macrophages indicated the activation of cells via NF-κB and MAPKs signaling pathways which was followed after CBF<SUB>2</SUB>-macrophage interaction through TLR4 and TLR2 receptors. The most potent macrophage activating polysaccharide, BCF<SUB>2</SUB>, contained a backbone of (1 → 6)-β-galactopyranosyl units branched at C-3.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Water soluble acidic polysaccharides were isolated and purified from <I>Boswellia carterii.</I> </LI> <LI> The molecular weights of polysaccharides ranged from 295.5 to 504.7 × 10<SUP>3</SUP> g/mol. </LI> <LI> Polysaccharides exhibited notable RAW264.7 macrophage stimulating activity. </LI> <LI> Macrophages were activated through NF-κB and MAPK pathways via TLR4 and TLR2. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>