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1.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 487-494, 2017.
Article in English | WPRIM | ID: wpr-812089

ABSTRACT

Lipopolysaccharides (LPS) contamination in herbal crude polysaccharides is inevitable. The present study was performed to explore the effect of polymyxin B on abolishing the influence of LPS contamination in mononuclear cells. LPS was pretreated with polymyxin B sulfate (PB) at different concentrations for 1, 5 or 24 h, and then used to stimulate RAW264.7 and mouse peritoneal macrophages (MPMs). The nitric oxide (NO) and tumor necrosis factor-α (TNF-α) in cell culture supernatant, as the indications of cell response, were assayed. Bupleurum chinensis polysaccharides (BCPs) with trace amount contamination of LPS was treated with PB. 30 μg·mL of PB, treating LPS (10 and 1 000 ng·mL in stimulating RAW264.7 and MPMs respectively) at 37 °C for 24 h, successfully abolished the stimulating effect of LPS on the cells. When the cells were stimulated with LPS, BCPs further promoted NO production. However, pretreated with PB, BCPs showed a suppression of NO production in MPMs and no change in RAW264.7. In the in vitro experiments, LPS contamination in polysaccharide might bring a great interference in assessing the activity of drug. Pretreatment with PB (30 μg·mL) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination (10 and 1 000 ng·mL).


Subject(s)
Animals , Mice , Bupleurum , Chemistry , Drug Contamination , Drugs, Chinese Herbal , Pharmacology , Lipopolysaccharides , Macrophages , Metabolism , Nitric Oxide , Metabolism , Polymyxin B , Pharmacology , Polysaccharides , Pharmacology , Tumor Necrosis Factor-alpha , Metabolism
2.
Chinese Journal of Natural Medicines (English Ed.) ; (6): 889-898, 2017.
Article in English | WPRIM | ID: wpr-812044

ABSTRACT

The present study aimed to investigate the effects of polysaccharides extracted from Bupleurum chinense DC (BCPs) on macrophage functions. In the in vivo experiment, 1 mL of 5% sodium thioglycollate was injected into the abdomen of the mice on Day 0 and macrophages were harvested on Day 4. The macrophages were cultured in plates and treated with different concentrations of BCPs and stimulus. Effects of BCPs on macrophage functions were assessed by chemotaxis assay, phagocytosis assay and Enzyme-Linked Immunosorbent Assay (ELISA). Our results showed the enhanced chemotaxis, phagocytosis and secretion of nitric oxide (NO) and inflammatory cytokines by macrophages when treated with BCPs. However, when chemotaxis and phagocytosis were up-regulated by complement components or opsonized particles, BCPs inhibited these effects. Also, the NO production induced by lipopolysaccharides (LPS) was suppressed by BCPs mildly. Moreover, BCPs had an inhibitory effect on the [Ca] elevation of macrophages. These results suggested that BCPs exerted modulatory effects on macrophage functions, which may contribute to developing novel approaches to treating inflammatory diseases.


Subject(s)
Animals , Mice , Bupleurum , Chemistry , Chemotaxis , Cytokines , Metabolism , Immunologic Factors , Pharmacology , Immunomodulation , Macrophages , Mice, Inbred BALB C , Nitric Oxide , Metabolism , Phagocytosis , Plant Extracts , Chemistry , Pharmacology , Plant Roots , Chemistry , Plants, Medicinal , Chemistry , Polysaccharides , Pharmacology
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