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1.1 Animals Balb / c mice, 6-7 weeks old, weight 20-22g,[link widoczny dla zalogowanych], Department of Basic Medicine, Jilin University Experimental Animal Center.



1 experimental material
Araliaceae Panax ginseng as a plant, sweet, bitter , cool, with a strong tonic, nourishing fluid, rather God puzzle and so on. Modern pharmacological studies have shown that Panax quinquefolius saponin with anti-virus,[link widoczny dla zalogowanych], anti-mutation, anti-arrhythmia, lower blood lipids, such as the role of lipid peroxidation [1]. Panax quinquefolius saponin for further study of immune regulation, the experimental observation of American ginseng leaf saponins on in vitro metabolism of mouse peritoneal macrophages and generation of NO concentration, IL  1 activity.


1.2 Drugs and reagents Panax quinquefolius saponin provided by the Institute of Chinese Medicine of Jilin Province: MTT, bacterial lipopolysaccharide (LPS), concanavalin A (ConA), Sigma; dimethyl sulfoxide sulfone (DMSO), Shantou, Guangdong Xilong Chemical Products; RPMI1640 medium, the United States GIBCO products; fetal calf serum (NBS) Beijing National Center for Biotechnology Development tripod provides; NO kit by Nanjing Jiancheng Bioengineering Institute; other reagents were of analytical grade.



Panax quinquefolius saponin on mouse peritoneal macrophages in immune function of


Chinese papers League finishing. Of: Ding Tao, Sheung Chi, Wen Fuchun, Zhang Wei, Zhang Dianwen, Jifeng Lan, Xuhui Bo



2.2 macrophages Metabolic Test 96 macrophage monolayer culture plates with different concentrations of reagent 10μl / hole, PBS negative control by adding balanced salt solution,[link widoczny dla zalogowanych], 5 duplicate wells for each concentration, and then each well adding 10% NBS  PRMI1640 medium 100μl, set 37 ℃ 5% CO2 incubator for 24h, MTT reagent added to each well 10μl, to continue to foster 4h, each hole by adding DMSO100μl, micro oscillator 10min, microplate reader at 570nm absorbance A measured value [3].

1.3 Instruments microplate reader, TECAN Austria products; CO2 incubator, Japan SANYO products; inverted microscope, Japan OLYMPUS products; TU  1800S spectrophotometer, Beijing General Instrument Co., S & P analysis Company.
2.3 macrophages to produce NO was determined by nitrate reductase. Single 24-well plates of macrophages, each hole by adding a final concentration of 10μg/ml of LPS0.5ml, to activate macrophages, while each hole by adding different concentrations of reagent 1ml, 6 duplicate wells for each concentration at 37 ℃ 5% CO2 incubator for 24h, culture supernatants obtained 0.5ml per well tested, and then press the NO kit instructions to join the appropriate reagents, at room temperature for 10min, 550nm wavelength spectrophotometer absorbance A measured value,[link widoczny dla zalogowanych], according to kit formula amount of NO formation [4].
2.1 mouse peritoneal macrophages of mice 6 Preparation of the abdomen after disinfection in the intraperitoneal injection of each serum-free RPMI1640 medium 3ml, 15min after the cervical dislocation mice were killed, 75% alcohol soaked 5min, into clean table, intra-abdominal fluid collection, 1000r/min centrifugal 5min, supernatant, PBS washed cells in balanced salt solution 3 times, with 10% NBS  RPMI1640 culture medium adjusted cell concentration to 106/ml, 24 well plate each well cell suspension 1ml (96 well plate cell suspension added per well 100μl), set to 37 ℃ 5% CO2 incubator for 4h, supernatant balance with PBS wash away the salt solution is not adherent cells, cultured macrophages single hole [2].
2 experiments


2.4 macrophages to produce IL  1 of thymus cells with value-added method. Single 24-well plates of macrophages, each hole by adding a final concentration of 10μg/ml of LPS0.5ml, to activate macrophages, while each hole by adding different concentrations of reagent 1ml, culture supernatants harvested after 48h, was induced of IL  1 crude, -20 ℃ saved under test. Preparation of 107/ml normal thymus cell suspension, added 96, each well 0.1ml, while adding different dilutions of IL  1 tested supernatant 0.1ml, final concentration 2μg/ml the ConA0.1ml, each a concentration of 3 re-holes, set 37 ℃ 5% CO2 incubator for 68h, the MTT reagent added to each well 10μl, to continue to foster 4h, each hole by adding DMSO100μl, micro oscillator 10min, microplate reader measuring absorbance at 570nm A value [5].
Key words Panax quinquefolius saponin mouse peritoneal macrophage nitric oxide in interleukin-1
Abstract Objective: To observe the stems and leaves of American ginseng saponins on activated mouse peritoneal macrophage metabolism and secretion of nitric oxide (NO), interleukin-1 (IL  1) activity. Methods: Different concentrations of ginseng stem saponins added in vitro cultured mouse peritoneal macrophages to NO kit NO content in the supernatant to MTT (MTT) colorimetric assay metabolism of macrophages and production of IL  1 activity. Results: Panax quinquefolius saponins can stimulate the metabolism of mouse peritoneal macrophages and generation of NO,[link widoczny dla zalogowanych], and NO production in a dose dependent relationship between the activity of IL  1 had no effect. Conclusion: Ginseng Leaves Extract can enhance the activity of mouse peritoneal macrophages may be Panax quinquefolius saponin regulating immune function in an important way. More articles related to topics：


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