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 m L of 5% sodium thioglycollate was injected into ...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 m L 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^(2+)]i 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.展开更多
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 mononuclea...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^(–1) of PB, treating LPS(10 and 1000 ng·mL^(–1) in stimulating RAW264.7 and MPMs respectively) at 37 ℃ 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^(–1)) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination(10 and 1 000 ng·mL^(–1)).展开更多
基金Supported by Innovation Alliance Project of Agricultural Science and Technology Innovation Fund of Hunan Province(2017LM0301)Science and Technology Innovation Project in Hunan Academy of Agricultural Sciences(2017JC55)Key Research&Development Project of Hunan(2016NK2180)~~
基金supported by the National Natural Science Foundation of China(Nos.30925042,81274165,and 81330089)the State Key Program for New Drugs from the Ministry of Science and Technology,China(No.2012ZX09301001-003)the Science and Technology Commission of Shanghai Municipality(Nos.10XD1405900 and 12JC1400800)
文摘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 m L 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^(2+)]i 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.
基金supported by the National Natural Science Foundation of China(Nos.81330089,30925042,and 81274165)the State Key Program for New Drugs from the Ministry of Science and Technology,China(No.2012ZX09301001-003)the Science and Technology Commission of Shanghai Municipality(Nos.10XD1405900 and 12JC1400800)
文摘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^(–1) of PB, treating LPS(10 and 1000 ng·mL^(–1) in stimulating RAW264.7 and MPMs respectively) at 37 ℃ 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^(–1)) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination(10 and 1 000 ng·mL^(–1)).
