体外细胞培养检验“方证对应”关系的方法学研究  

Methodological Study on Effect of Traditional Chinesc Medicine Based on Syndrome Differentiation on Natural Killer Cell activity In Vitro

作　　者：朱唯芳[1] 黄海茵[1] 郭映华[1] 

机构地区：[1]上海市肺科医院,200433

出　　处：《中华临床医学卫生杂志》2006年第3期5-7,共3页China Journal of Clinical Medicine Hygiene

摘　　要：目的通过在体外细胞培养中加入中药复方粗提制剂对具有典型的中医证型的肺癌患者的外周血淋巴细胞NK活性的影响，探索能否在体外实验中反映“方证对应”关系。方法以水煎醇沉法制备两种中药复方（益气养阴和健脾化痰）的粗提制剂，取具有典型的中医证型（气阴两虚或脾虚痰湿证型）的肺癌患者的外周血淋巴细胞，将同一个患者的标本分为四个体外实验组，即空白对照组，中药5mg／ml组及10mg／ml组，自细胞介素2（250^u/ml，阳性对照）组。按“辨证论治”原则加入相应的中药复方制剂。体外培养22h或94h，以MTT法检测各组的NK活性。结果无论培养22h或94h，中药组的NK活性与对照组比较皆无显著性差异（P〉0．05），而白细胞介素2（阳性对照）组的NK括性均明显升高（P〈0．05）。结论两种中药复方制剂都未能在体外反应出“方证对应”的效果，说明中药粗提制剂直接用于体外细胞培养，其结果的可靠性差。Objective To investigate the effect of total extracts of composite recipes of traditional Chinese medicine（TCM） based on syndrome diffcrrentiation on natural killer cell（NK） activity in vitro. Methods The total extracts of two composite recipes（replenishing Qi and nourishing Yin;invigorating Spleen and eliminating Phlegm） were prepared by the method of water decocting and ethanol extracting.The patients of lung cancer who were typical in syndrome differrentiation typing（deficiency of both Qi and Yin,or Spleen deficiency with Phlegm stagnancy） were selected and their peripheral blood mononuclear cells isolated,the sample of every patient was further divided into four groups：the control group,TCM 5mg/ml group,TCM 10mg/ml group, Interleukin- 2（250^u/ml）group. extracts of composite recipes ,according to syndrome differrcntiation typing ,were added to the cell culture of the TCM groups.After 22 or 94 hours of culture,the NK activity of each group were determined with colorimetric MTT assay. Results No significant differrence was noted between the NK activity of the TCM group and the control group（P〉0.05）.However, the NK activity of the Interleukin-2 group was significantly higher than the control group either after 22 or 94 hours.Conclusion The effect of total extracts of both of the two composite recipes in vitro didn＇t accord with the effect of them in vivo. Total extracts of composite recipes of TCM isn＇t suitable for in vitro tissue culture,for the method is susceptible to a lot of intcrfereing factors and its result unreliable.

关 键 词：辨证论治 中药复方 体外 组织培养 自然杀伤细胞 

分 类 号：R-33[医药卫生]