评价中药寒热药性的实验方法研究

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。

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

Objective T o set up the experimental method of evaluating the cold and hot characteristicof traditional Chinese medicine (T CM) through investigating the different effects of T CM with cold andhot characteristic on the growth and proliferation of human breast cancer cell line MCF-7 [WT BX] in vitro.[WTBZ] Methods.MTT Assay was used to investigate the effect of four herbal drugs of T CM with coldand cool characteristic (cold drug) and five herbal drugs of TCM with hot and warm characteristic (hotdrug) on the growth and proliferation of MCF-7 [WTBX] in vitro. [WTBZ] Morphological changes ofMCF-7 treated with cold and hot herbal drugs were observed through inverted microscope. T rypan-bluestaining was used to analyze the cytotoxicity of herbal drugs we have chosen. Results .The cold drugs([WT BX] Coptis chinensis, Polygonum cuspidatum, Phyllostachys nigra [WTBZ] var. [WT BX] henonis,and Prunella vulgaris[WT BZ] ) could inhibit the growth and prolif eration of MCF-7 in the concentra-tion range of 50.800. g/ mL and the inhibitory ef f ect is dependent on the concentration. The hot drugs([WT BX ] Zingiber of f icinale[WTBZ] and [WT BX ] Piper nigrum[WT BZ] ) could promote the growthand p rolif eration of MCF-7 in a concentration-dep endent manner. The concentration range is 50-800.g/ mL. The promotive ef f ect of [WT BX ] Z. of f icinale[WT BZ] could increase along with the increas-ing of the drug concentration. The promotive ef f ect of [WT BX ] P. nigrum[WT BZ] could increasef irstly but then decrease along with the increasing of the drug concentration. The rest of the herbal drugwith hot characteristic could promote the growth and prolif eration of MCF-7 in the low concentrationrange ([WT BX ] Cinnamomum cassia [WT BZ] and [WT BX ] Zanthoxylum bungeanum[WTBZ] 50-200.g/ mL, [WT BX ] Curculigo orchioides 50-400 /g/ mL) , the promotive effect could increase along withthe increasing of the concentration; but inhibit in the high concentration range ([WTBX] C. cassia. [WT-BZ] and [WTBX] Z. bungeanum [WT BZ] 400-800. g/ mL, [WT BX] C. orchioides[WT BZ] 600. 800 ?g/mL), the inhibitory effect could increase along with the increasing of the concentration. Morphological ob-servation showed that the cold drugs in the concentration range we have chosen could decrease the densityof MCF-7 and condense the cell, while hot drugs make MCF-7 grow well and the density rise in a certainconcentration range. Trypan-blue staining showed that the drugs we have chosen had no cytotoxicity onMCF-7. ConclusionAll these results show that the method in this paper may be used to evaluate the coldand hot characteristic of T CM.