摘要
目的：观察黄芪桂枝五物汤中皂苷对机体免疫功能抑制的影响。方法：选取3~7周龄,清洁级昆明种小鼠,体质量（18±4）g,雌雄各半,取小鼠8只,雌雄各4只,脱颈处死,取脾脏,放入细胞筛中,用玻璃注射器内塞在3 m L PBS中研磨脾脏,离心混悬液,沉淀物中加2 m L蒸馏水充分破碎混悬液中红细胞,约半分钟后加入1.8%氯化钠溶液2 m L,离心,磷酸缓冲液洗涤细胞,计数,调浓度为1.0×106·m L-1。在细胞培养板中,实验组加入100μL细胞悬液,从黄芪桂枝五物汤中提取其有效成分皂苷,作用于小鼠脾细胞,取皂苷溶液10μL,按设计浓度分别进行实验,对照组加10μL缓冲液。采用血球计数板计数细胞数以判断其增殖活性,酶联免疫吸附试验（ELISA）测细胞因子（IL-2,IFN-γ）含量,流式细胞仪（FCM）检测细胞表型变化情况。结果：实验组淋巴细胞增殖活性优于对照组（P〈0.05）;实验组IL-2、IFN-γ浓度优于对照组,且随皂苷浓度的增加而降低,在一定浓度范围内成剂量依赖关系;细胞表型分析显示,皂苷既能抑制T淋巴细胞增殖,也能抑制B淋巴细胞增殖和细胞因子IL-2、IFN-γ的分泌。结论：黄芪桂枝五物汤中皂苷能抑制T淋巴细胞增殖、B淋巴细胞增殖和细胞因子分泌,改善IL-2、IFN-γ浓度,且随皂苷浓度的增加而降低,在一定浓度范围内成剂量依赖关系。
Objective： To observe the influence of the saponin in Astragalus and Cinnamon Twig Five Substances Decoction on the body immunity inhibition. Methods： Select 8 clean Kunming Mice of 3- 7 months old with body mass of（ 18 ± 4） g,4 female mice and 4 male mice,take off the neck to death,take the spleen and put into the cell strainer,grind the spleen with glass syringe plug in 3 m L PBS,centrifuge the suspension,add 2ml distilled water in the precipitate to well break the red blood cells in the suspention,add 2m L 1. 8% sodium chloride solution,centrifuge,clean the cells with phosphate bugger,count the number,and regulate the concentration to 1. 0 × 106·m L- 1. In the cell culture plate,add 100 ul cell suspension in experiment group. Extract the effective constituent in the Astragalus and Cinnamon Twig Five Substances Decoction to act on the spleen cells of mice,take 10 ul saponin solution,conduct experiment based on the designed concentration and add 10 ul buffer solution to control group. Judge the proliferative activity based on the the number of the cells with blood counting chamber,determine the content of IL-2 and IFN-γ by LISA,detect the changes of the cell phenotype with FCM. Results： The proliferative activity in experiment group was better than that of control group（ P 〈 0. 05）. The concentration of IL-2 and IFN-γ in experiment group was better than that of control group and decreased with the increase of the concentration of saponin; among certain concentration range,there was dosage dependency relationship. The analysis of cell phenotypic analysis showed that saponin can not only inhibit the proliferation of T lymphocytes but also suppress the proliferation of B lymphocytes and the secretion of IL-2 and IFN-γ. Conclusion： The saponin in Astragalus and Cinnamon Twig Five Substances Decoction can inhibit the proliferation of T lymphocytes,B lymphocytes and cytokines,improve the concentration of IL-2 and IFN-γ,and decreased with the increase of the concentration of saponin; among cert
出处
《河南中医》
2016年第3期479-481,共3页
Henan Traditional Chinese Medicine
基金
唐山市科学与研究计划指令课题(编号:13130253z)
关键词
黄芪桂枝五物汤
皂苷
细胞免疫
体液免疫
细胞因子
Astragalus and Cinnamon Twig Five Substances Decoction
saponin
cell immunity
humoral immunity
cytokines
