评价中药寒热药性的实验方法研究

Experimental method for evaluating cold and hot characteristic of traditional Chinese medicine

目的通过考察寒凉药与温热药对人乳腺癌细胞MCF-7体外生长增殖的影响来建立评价中药寒热药性的实验方法。方法用噻唑蓝比色法(MTT法)考察4种寒凉药和5种温热药对人乳腺癌细胞MCF-7体外生长增殖的影响,倒置显微镜观察寒凉药、温热药对细胞形态特征的影响,台盼蓝染色法分析所选寒凉药和温热药的细胞毒性作用。结果寒凉药(50～800μg/mL黄连、虎杖、竹叶、夏枯草)抑制MCF-7生长增殖,随质量浓度增大抑制作用增强。温热药干姜和白胡椒(50～800μg/mL)促进MCF-7生长增殖,干姜的促进作用随质量浓度增大而增强,白胡椒的促进作用先增大后减小。其余温热药低质量浓度(肉桂与花椒50～200μg/mL、仙茅50～400μg/mL)促进MCF-7生长增殖,促进作用随质量浓度增大而增强;高质量浓度(肉桂与花椒400～800μg/mL、仙茅600～800μg/mL)抑制MCF-7生长增殖,抑制作用随质量浓度增大而增强。形态学观察表明寒凉药(黄连、虎杖、竹叶、夏枯草)在所选质量浓度范围内使MCF-7密度减小,细胞固缩变圆;温热药(干姜、白胡椒、肉桂、仙茅、花椒)在各自起促进作用的质量浓度范围内使MCF-7密度增大,生长旺盛。台盼蓝染色表明各寒凉药与温热药对MCF-7没有细胞毒性作用。结论本方法有可能用于中药寒热药性的实验评价。

Objective To set up the experimental method of evaluating the cold and hot characteristic of traditional Chinese medicine （TCM） through investigating the different effects of TCM with cold and hot characteristic on the growth and proliferation of human breast cancer cell line MCF-7 in vitro. Methods MTT Assay was used to investigate the effect of four herbal drugs of TCM with cold and cool characteristic （cold drug） and five herbal drugs of TCM with hot and warm characteristic （hot drug） on the growth and proliferation of MCF-7 in vitro. Morphological changes of MCF-7 treated with cold and hot herbal drugs were observed through inverted microscope. Trypan-blue staining was used to analyze the cytotoxicity of herbal drugs we have chosen. Results The cold drugs （Coptis chinensis, Polygonum cuspidatum, Phyllostachys nigra var. henonis, and Prunella vulgaris） could inhibit the growth and proliferation of MCF-7 in the concentration range of 50-800 μg/mL and the inhibitory effect is dependent on the concentration. The hot drugs （Zingiber officinale and Piper nigrum） could promote the growth and proliferation of MCF-7 in a concentration-dependent manner. The concentration range is 50-800μg/mL. The promotive effect of Z. officinale could increase along with the increasing of the drug concentration. The promotive effect of P. nigrum could increase firstly but then decrease along with the increasing of the drug concentration. The rest of the herbal drug with hot characteristic could promote the growth and proliferation of MCF-7 in the low concentration range （Cinnamomum cassia and Zanthoxylum bungeanum 50-200μg/mL, Curculigo orchioides 50-400μg/mL）, the promotive effect could increase along with the increasing of the concentration; but inhibit in the high concentration range （C. cassia, and Z. bungeanum 400-800μg/mL, C. orchioides 600-800μg/mL）, the inhibitory effect could increase along with the increasing of the concentration. Morphological observation showed that the cold drugs in the concentration range we have chosen could decrease the density of MCF-7 and condense the cell, while hot drugs make MCF-7 grow well and the density rise in a certain concentration range. Trypan-blue staining showed that the drugs we have chosen had no cytotoxicity on MCF-7. Conclusion All these results show that the method in this paper may be used to evaluate the cold and hot characteristic of TCM.