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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| Format: | Article |
| Language: | English |
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Kafkas University, Faculty of Veterinary Medicine
2025-05-01
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| Series: | Kafkas Universitesi Veteriner Fakültesi Dergisi |
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| Online Access: | https://vetdergikafkas.org/pdf.php?id=3190 |
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| author | Guangpei XU Yanfei HE Ping JIANG Lei YANG Jianwen CAO Chuanbo SUN |
| author_facet | Guangpei XU Yanfei HE Ping JIANG Lei YANG Jianwen CAO Chuanbo SUN |
| author_sort | Guangpei XU |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-4513f0d37f3d4d67bad0a579c8e1f602 |
| institution | DOAJ |
| issn | 1309-2251 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Kafkas University, Faculty of Veterinary Medicine |
| record_format | Article |
| series | Kafkas Universitesi Veteriner Fakültesi Dergisi |
| spelling | doaj-art-4513f0d37f3d4d67bad0a579c8e1f6022025-08-20T03:23:51ZengKafkas University, Faculty of Veterinary MedicineKafkas Universitesi Veteriner Fakültesi Dergisi1309-22512025-05-0131336737510.9775/kvfd.2024.335923190Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathwaysGuangpei XU0Yanfei HE1Ping JIANG2Lei YANG3Jianwen CAO4Chuanbo SUN5West Anhui University, College of Biological and Pharmaceutical Engineering, 237000, CHINAWest Anhui University, College of Biological and Pharmaceutical Engineering, 237000, CHINAWest Anhui University, College of Biological and Pharmaceutical Engineering, 237000, CHINAWest Anhui University, College of Biological and Pharmaceutical Engineering, 237000, CHINAWest Anhui University, College of Biological and Pharmaceutical Engineering, 237000, CHINAWest Anhui University, College of Biological and Pharmaceutical Engineering, 237000, CHINATo 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.https://vetdergikafkas.org/pdf.php?id=3190dendrobium officinale polysaccharideraw264.7 macrophagescytokinesimmunomodulation |
| spellingShingle | Guangpei XU Yanfei HE Ping JIANG Lei YANG Jianwen CAO Chuanbo SUN Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways Kafkas Universitesi Veteriner Fakültesi Dergisi dendrobium officinale polysaccharide raw264.7 macrophages cytokines immunomodulation |
| title | Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways |
| title_full | Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways |
| title_fullStr | Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways |
| title_full_unstemmed | Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways |
| title_short | Dendrobium officinale polysaccharide regulates the immune function of raw264.7 cells via the nf-κb and mapk signaling pathways |
| title_sort | dendrobium officinale polysaccharide regulates the immune function of raw264 7 cells via the nf κb and mapk signaling pathways |
| topic | dendrobium officinale polysaccharide raw264.7 macrophages cytokines immunomodulation |
| url | https://vetdergikafkas.org/pdf.php?id=3190 |
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