Modulatory effects of the acid polysaccharide fraction from one of anamorph of Cordyceps sinensis on Ana-1 cells.
BACKGROUND
Cordyceps sinensis has been used as a precious herbal medicine for thousands of years in China. Its polysaccharide fraction has been confirmed possessing immunomodulatory function and we have reported the acid polysaccharide fraction (APSF), from an anamorph of C. sinensis, has stimulating activity on macrophages. The mechanism still needs to be further elucidated.


METHODS
In order to investigate the effects of APSF on macrophage's phenotypes, Ana-1 mouse macrophages were polarized to M2 phenotype by culturing the cells with culture supernatant of H22 cells. M2 phenotype was determined by measuring the expression of TNF-α and checking cell surface markers mannose receptor (MR) and scavenger receptor (SR). After cultured with H22 supernatant for 72 h, the TNF-α level of Ana-1 cells was decreased while the SR and MR expressions were up-regulated, suggesting that Ana-1 cells were polarized towards M2 macrophages. Then the effects of APSF on M2 macrophages were investigated by measuring mRNA levels of TNF-α, inducible nitric oxide synthase (iNOS), IL-12 and IL-10. Nuclear NF-κB was detected by Western blotting.


RESULTS
APSF treatment increased the expressions of TNF-α, IL-12 and iNOS, and reduced the expression of IL-10 of Ana-1 cells. Besides, the expressions of SR and MR were down-regulated by APSF. And the result of Western blotting showed NF-κB level was decreased in M2 macrophages and up-regulated after APSF treatment.


CONCLUSIONS
APSF may convert M2 macrophages to M1 phenotype by activating NF-κB pathway.
Jiangsu Key Laboratory of Molecular Medicine, Medical School and State Key Laboratory of Analytical Chemistry for Life Science, Nanjing University, Nanjing 210093, People's Republic of China.
Weixia
W
Chen
Fengjiao
F
Yuan
Kongcheng
K
Wang
Dan
D
Song
Weiyun
W
Zhang
D000818:Animals; D018122:B7-1 Antigen / Q000378:metabolism; D051940:B7-2 Antigen / Q000378:metabolism; D002460:Cell Line; D045744:Cell Line, Tumor; D032661:Cordyceps; D016207:Cytokines / Q000235:genetics; D003911:Dextrans / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000378:metabolism; D007155:Immunologic Factors / Q000494:pharmacology*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics; D010641:Phenotype; D011134:Polysaccharides / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism
D018122:B7-1 Antigen; D051940:B7-2 Antigen; C497965:Cd86 protein, mouse; D016207:Cytokines; D003911:Dextrans; D007155:Immunologic Factors; D016328:NF-kappa B; D011134:Polysaccharides; D012333:RNA, Messenger; C015219:fluorescein isothiocyanate dextran; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D016650:Fluorescein-5-isothiocyanate