摘要
研究中药粗提剂影响体外淋巴细胞转化试验时,常因所含杂质多浓度偏高或细胞收集不全,使 ~3H 标记的胸腺嘧啶脱氧核糖苷(~3H-TdR)掺入受阻或细胞丢失而致 cpm 偏低,造成假阴性。加用镜检观察细胞增殖为指标时,它受上述因素干扰较小,可有很高的可信性、重复性,较宽的检出浓度范围,较易检测成功,以及可多人同时评定使其客观性较强等特点,它与~3H—TdR 掺入指标两者互相参比,可获客观确切结果。增强细胞收集时所用负压.延长冲吸时间、镜验收集是否完全等,可防止中药粗提剂增强细胞粘附培养板孔面而致收集不全之弊。
When in vitro lymphocyte transformation test was used to investigate the immunologic effects of crude herbal extracts,false negative results of ~3H-TdR uptake usually were obtained due to the high content of impurities,higher concentration of drugs or incomplete harvest of cells.Microscopic examination of cell growth to be additional index proved very useful because of being less interfered by factor mentioned above.Cell growth examination has high reliability,reproducibility,wider range of suitable drug concentration detectable(20～40μg crude extract/well)and high objectivity of being capably examined by several observers for comparison.Therefore both ~3H-TdR uptake and cell growth examination should be adopted for indications of this test to obtain accurate objective results.In case of increasing adhesion of cells with surfaces of plate wells by crude herbal extract it is necessary to elevate negative pressure(700 mmHg)and prolong the time of wash-suction (1.5～2min)during cell harvesting.
基金
中国药材公司科研资助课题
