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      Polymyxin B as an inhibitor of lipopolysaccharides contamination of herb crude polysaccharides in mononuclear cells

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          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 1 000 ng·mL –1 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 –1) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination (10 and 1 000 ng·mL –1).

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          Most cited references 23

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          Two physically, functionally, and developmentally distinct peritoneal macrophage subsets.

          The peritoneal cavity (PerC) is a unique compartment within which a variety of immune cells reside, and from which macrophages (MØ) are commonly drawn for functional studies. Here we define two MØ subsets that coexist in PerC in adult mice. One, provisionally called the large peritoneal MØ (LPM), contains approximately 90% of the PerC MØ in unstimulated animals but disappears rapidly from PerC following lipopolysaccharide (LPS) or thioglycolate stimulation. These cells express high levels of the canonical MØ surface markers, CD11b and F4/80. The second subset, referred to as small peritoneal MØ (SPM), expresses substantially lower levels of CD11b and F4/80 but expresses high levels of MHC-II, which is not expressed on LPM. SPM, which predominates in PerC after LPS or thioglycolate stimulation, does not derive from LPM. Instead, it derives from blood monocytes that rapidly enter the PerC after stimulation and differentiate to mature SPM within 2 to 4 d. Both subsets show clear phagocytic activity and both produce nitric oxide (NO) in response to LPS stimulation in vivo. However, their responses to LPS show key differences: in vitro, LPS stimulates LPM, but not SPM, to produce NO; in vivo, LPS stimulates both subsets to produce NO, albeit with different response patterns. These findings extend current models of MØ heterogeneity and shed new light on PerC MØ diversity, development, and function. Thus, they introduce a new context for interpreting (and reinterpreting) data from ex vivo studies with PerC MØ.
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            Functional macrophage cell lines transformed by Abelson leukemia virus.

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              Binding of polymyxin B to the lipid A portion of bacterial lipopolysaccharides.


                Author and article information

                Chinese Journal of Natural Medicines
                20 July 2017
                : 15
                : 7
                : 487-494
                1Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China
                2Department of Pharmacognosy, School of Pharmacy, Fudan University, Shanghai 201203, China
                Author notes
                *Corresponding author: ZHANG Yun-Yi, Tel: 86-21-51980050, zhcheng@ 123456shmu.edu.cn ; CHEN Dao-Feng, Tel: 86-21-51980135, dfchen@ 123456shmu.edu.cn

                These authors have no conflict of interest to declare.

                Copyright © 2017 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.
                Funded by: National Natural Science Foundation of China
                Award ID: 81330089
                Award ID: 30925042
                Award ID: 81274165
                Funded by: Ministry of Science and Technology
                Award ID: 2012ZX09301001-003
                Funded by: Science and Technology Commission of Shanghai Municipality
                Award ID: 10XD1405900
                Award ID: 12JC1400800
                This work was 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), and the Science and Technology Commission of Shanghai Municipality (Nos. 10XD1405900 and 12JC1400800).


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