Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways
To investigate the immunomodulatory effects of Dendrobium officinale polysaccharide (DOP) on RAW264.7 macrophages. DOP was purified from Dendrobium officinale using water extraction-alcohol precipitation and gel column chromatography. The molecular weight distribution of the polysaccharide was analy...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Kafkas University, Faculty of Veterinary Medicine
2025-05-01
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| Series: | Kafkas Universitesi Veteriner Fakültesi Dergisi |
| Subjects: | |
| Online Access: | https://vetdergikafkas.org/pdf.php?id=3190 |
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| Summary: | To investigate the immunomodulatory effects of Dendrobium officinale polysaccharide
(DOP) on RAW264.7 macrophages. DOP was purified from Dendrobium officinale using
water extraction-alcohol precipitation and gel column chromatography. The molecular
weight distribution of the polysaccharide was analyzed by high-performance gel
permeation chromatography (HPGPC). The effects of DOP on RAW264.7 cells were
evaluated as follows: cell viability was assessed via MTT assay to screen appropriate
treatment concentrations; ELISA was used to detect the secretion of inflammatory
cytokines (IL-10, IL-12 p40, and IL-6); RT-qPCR was employed to measure the
expression of inflammatory genes (IL-1β, IL-10, IL-12 p40, and IL-6); and Western
blotting was performed to analyze the expression of P65, phosphorylated P65 (p-P65),
JNK, phosphorylated JNK (p-JNK), ERK, and phosphorylated ERK (p-ERK). The
purified DOP exhibited a yield of 8.24% and a total sugar content of 95.33±3.11%.
HPGPC analysis revealed three peaks corresponding to molecular weights of 1.89 x
10⁵ Da, 549 Da, and 211 Da. DOP showed no cytotoxicity toward RAW264.7 cells at
concentrations ranging from 3.125 to 400 μg/mL. All tested concentrations of DOP
significantly enhanced the mRNA expression of IL-1β, IL-10, IL-12 p40, and IL-6
(P<0.05, P<0.01, P<0.001) and promoted the secretion of IL-10, IL-12 p40, and IL-6 in
a dose-dependent manner. Regarding the NF-κB and MAPK signaling pathways, DOP
markedly increased the expression of p-JNK, p-ERK, and p-P65 proteins (P<0.001), with
sustained effects observed for up to 120 minutes (P<0.001). DOP activates RAW264.7
cells by promoting inflammatory gene expression, stimulating cytokine secretion, and
triggering the NF-κB and MAPK pathways, thereby exerting immunomodulatory
effects. |
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| ISSN: | 1309-2251 |