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Immunomodulating activity of Nymphaea rubra Roxb. extracts: activation of rat dendritic cells and improvement of the T(H)1 immune response.

Cheng JH, Lee SY, Lien YY, Lee MS, Sheu SC - Int J Mol Sci (2012)

Bottom Line: CD80/86 (87.16% ± 8.49%) and MHC class II (52.01% ± 10.11%) expression levels were significantly up-regulated by this treatment compared to the controls (65.45% ± 0.97% and 34.87% ± 1.96%).In parallel, endocytosis was also reduced (167.94% ± 60.59%) after treatment with 25 μg/mL of NR-PS as measured by the medium fluorescence intensity compared to the control (261.67% ± 47.26%).In conclusion, NR-PS exhibits stimulatory effects on rat DCs and promotes the secretion of T(H)1 cytokines.

View Article: PubMed Central - PubMed

Affiliation: The Department of Nursing, Shu Zen College of Medicine and Management, Kaohsiung 821, Taiwan; E-Mail: cjaiho@yahoo.com.tw.

ABSTRACT
Polysaccharides play a key role in enhancing immune function and facilitating cellular communication. Here, we purified Nymphaea rubra Roxb. polysaccharides (NR-PS) by treating them with pullulanase. They were then cultured with immature dendritic cells (DCs) derived from rat bone marrow hematopoietic cells (BMHCs). After treatment with bioactive NR-PS with a degree of polymerization (DP) value of 359.8, we found that the DCs underwent morphological changes indicative of activation. CD80/86 (87.16% ± 8.49%) and MHC class II (52.01% ± 10.11%) expression levels were significantly up-regulated by this treatment compared to the controls (65.45% ± 0.97% and 34.87% ± 1.96%). In parallel, endocytosis was also reduced (167.94% ± 60.59%) after treatment with 25 μg/mL of NR-PS as measured by the medium fluorescence intensity compared to the control (261.67% ± 47.26%). Furthermore, the DCs after treatment with 25 μg/mL NR-PS showed increased IL-12 (102.09 ± 10.16 to 258.78 ± 25.26 pg/mL) and IFN-γ (11.76 ± 0.11 to 15.51 ± 1.66 pg/mL) secretion together with reduced IL-10 secretion (30.75 ± 3.35 to 15.37 ± 2.35 pg/mL), which indicates a T(H)1 immune response. In conclusion, NR-PS exhibits stimulatory effects on rat DCs and promotes the secretion of T(H)1 cytokines. Taken together, our studies are the first to show that NR-PS is an immunomodulator affecting the maturation and functioning of DCs.

No MeSH data available.


(A) The flower of Nymphaea rubra Roxb.; (B) Schematic of a longitudinal section through a quartered portion of the flower, (sep) sepals, (pet) petals, (s) stamen, (ca) carpellary appendage, (c) carpel; (C) The carpel of Nymphaea rubra Roxb.
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f1-ijms-13-10722: (A) The flower of Nymphaea rubra Roxb.; (B) Schematic of a longitudinal section through a quartered portion of the flower, (sep) sepals, (pet) petals, (s) stamen, (ca) carpellary appendage, (c) carpel; (C) The carpel of Nymphaea rubra Roxb.

Mentions: The carpels of Nymphaea rubra Roxb. were collected (Figure 1C) and ground to give a crude extract. We then prepared both crude and hydrolyzed Nymphaea rubra Roxb. extracts and analyzed their composition. As shown in Table 1, the yield of crude extraction was 1.43% (wet weight). As previously reported [27,28], β-1,3-d-glucan has important biological functions and therefore pullulanase was added to the crude extract to hydrolyze any α-1,6 linkages present and to increase the solubility of the crude extract without breaking the β-1,3-glycosidic linkages. The yield of the hydrolyzed sample was 42.00% (dry weight) relative to the crude extract. The total sugar content and reducing sugar content of the hydrolyzed extract were 51.3% and 2.45%, respectively. The total protein content of the crude and hydrolyzed extracts were 7.84% and 5.28% respectively. The DP value of the hydrolyzed extract was found to be at 359.8 kDa by Glass Capillary Viscometer with a degree of branching (DB) value of 0.36. Bohn and BeMiller reported that the most active polysaccharide polymers have a high molecular weight (up to 100 kDa) and a DB between 0.2 and 0.5 [29]. Ohno et al. reported that the polysaccharide fractions prepared from cultured A. blazei containing a highly branched 1,3-β-glucan segment are the most active and have the highest antitumor activity and immunomodulary effects [30]. The Nymphaea rubra Roxb. polysaccharides (NR-PS) preparation used in this study had a DB of 0.36 and was within the bioactive range of polysaccharide polymers.


Immunomodulating activity of Nymphaea rubra Roxb. extracts: activation of rat dendritic cells and improvement of the T(H)1 immune response.

Cheng JH, Lee SY, Lien YY, Lee MS, Sheu SC - Int J Mol Sci (2012)

(A) The flower of Nymphaea rubra Roxb.; (B) Schematic of a longitudinal section through a quartered portion of the flower, (sep) sepals, (pet) petals, (s) stamen, (ca) carpellary appendage, (c) carpel; (C) The carpel of Nymphaea rubra Roxb.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472710&req=5

f1-ijms-13-10722: (A) The flower of Nymphaea rubra Roxb.; (B) Schematic of a longitudinal section through a quartered portion of the flower, (sep) sepals, (pet) petals, (s) stamen, (ca) carpellary appendage, (c) carpel; (C) The carpel of Nymphaea rubra Roxb.
Mentions: The carpels of Nymphaea rubra Roxb. were collected (Figure 1C) and ground to give a crude extract. We then prepared both crude and hydrolyzed Nymphaea rubra Roxb. extracts and analyzed their composition. As shown in Table 1, the yield of crude extraction was 1.43% (wet weight). As previously reported [27,28], β-1,3-d-glucan has important biological functions and therefore pullulanase was added to the crude extract to hydrolyze any α-1,6 linkages present and to increase the solubility of the crude extract without breaking the β-1,3-glycosidic linkages. The yield of the hydrolyzed sample was 42.00% (dry weight) relative to the crude extract. The total sugar content and reducing sugar content of the hydrolyzed extract were 51.3% and 2.45%, respectively. The total protein content of the crude and hydrolyzed extracts were 7.84% and 5.28% respectively. The DP value of the hydrolyzed extract was found to be at 359.8 kDa by Glass Capillary Viscometer with a degree of branching (DB) value of 0.36. Bohn and BeMiller reported that the most active polysaccharide polymers have a high molecular weight (up to 100 kDa) and a DB between 0.2 and 0.5 [29]. Ohno et al. reported that the polysaccharide fractions prepared from cultured A. blazei containing a highly branched 1,3-β-glucan segment are the most active and have the highest antitumor activity and immunomodulary effects [30]. The Nymphaea rubra Roxb. polysaccharides (NR-PS) preparation used in this study had a DB of 0.36 and was within the bioactive range of polysaccharide polymers.

Bottom Line: CD80/86 (87.16% ± 8.49%) and MHC class II (52.01% ± 10.11%) expression levels were significantly up-regulated by this treatment compared to the controls (65.45% ± 0.97% and 34.87% ± 1.96%).In parallel, endocytosis was also reduced (167.94% ± 60.59%) after treatment with 25 μg/mL of NR-PS as measured by the medium fluorescence intensity compared to the control (261.67% ± 47.26%).In conclusion, NR-PS exhibits stimulatory effects on rat DCs and promotes the secretion of T(H)1 cytokines.

View Article: PubMed Central - PubMed

Affiliation: The Department of Nursing, Shu Zen College of Medicine and Management, Kaohsiung 821, Taiwan; E-Mail: cjaiho@yahoo.com.tw.

ABSTRACT
Polysaccharides play a key role in enhancing immune function and facilitating cellular communication. Here, we purified Nymphaea rubra Roxb. polysaccharides (NR-PS) by treating them with pullulanase. They were then cultured with immature dendritic cells (DCs) derived from rat bone marrow hematopoietic cells (BMHCs). After treatment with bioactive NR-PS with a degree of polymerization (DP) value of 359.8, we found that the DCs underwent morphological changes indicative of activation. CD80/86 (87.16% ± 8.49%) and MHC class II (52.01% ± 10.11%) expression levels were significantly up-regulated by this treatment compared to the controls (65.45% ± 0.97% and 34.87% ± 1.96%). In parallel, endocytosis was also reduced (167.94% ± 60.59%) after treatment with 25 μg/mL of NR-PS as measured by the medium fluorescence intensity compared to the control (261.67% ± 47.26%). Furthermore, the DCs after treatment with 25 μg/mL NR-PS showed increased IL-12 (102.09 ± 10.16 to 258.78 ± 25.26 pg/mL) and IFN-γ (11.76 ± 0.11 to 15.51 ± 1.66 pg/mL) secretion together with reduced IL-10 secretion (30.75 ± 3.35 to 15.37 ± 2.35 pg/mL), which indicates a T(H)1 immune response. In conclusion, NR-PS exhibits stimulatory effects on rat DCs and promotes the secretion of T(H)1 cytokines. Taken together, our studies are the first to show that NR-PS is an immunomodulator affecting the maturation and functioning of DCs.

No MeSH data available.