均匀设计法优选肝豆汤的效应中药配伍——以TX小鼠神经细胞内Bcl-2、Bax蛋白表达水平及血清和脑组织中铜含量为指标  

A Study of Compatibility of Effective Traditional Chinese Medicine Components in Gandou Decoction Based on Uniform Design—Indicated by Protein Expression of Bcl-2 and Bax in Nerve Cells and Content of Copper in Serum and Brain Tissue of TX Mice

作　　者：汪世靖 赵雯[1,2] 陈林[1,2] 董健健[1,2] 程楠[2] 韩永升[2] 胡文彬[2] 韩咏竹[2] 

机构地区：[1]安徽中医药大学研究生部,安徽合肥230038 [2]安徽中医药大学神经病学研究所附属医院,安徽合肥230061

出　　处：《安徽中医药大学学报》2015年第5期69-74,共6页Journal of Anhui University of Chinese Medicine

基　　金：安徽省科技攻关计划项目(12010402114);国家自然科学基金项目(81473535)

摘　　要：目的基于均匀设计法筛选中药复方肝豆汤保护TX小鼠神经细胞的效应中药。方法基于"6因子11水平表"均匀设计法,以神经细胞内Bcl-2、Bax蛋白表达水平及血清和脑组织中铜含量为指标,以TX小鼠为Wilson病动物模型,采用多元逐步回归分析法优选肝豆汤中6种药物(大黄、姜黄、黄连、泽泻、金钱草、三七)的效应中药配伍。采用Western blot法以及等离子质谱法分别测定细胞凋亡相关蛋白Bcl-2、Bax和血清、脑组织铜含量。结果与对照组比较,TX小鼠脑组织中Bcl-2表达水平显著降低(P<0.05),Bax蛋白表达水平显著升高(P<0.05),脑组织铜含量显著升高(P<0.05),血清铜含量显著降低(P<0.05)。多元逐步回归分析结果显示,姜黄和大黄分别进入血清铜含量及脑组织铜含量的回归方程,黄连和泽泻均进入脑组织Bcl-2和Bax表达水平的回归方程,姜黄和金钱草分别进入神经细胞Bcl-2/Bax的回归方程,三七均未进入上述指标的回归方程。结论肝豆汤中大黄、姜黄、黄连、泽泻、金钱草配伍组合,可通过多靶点保护TX小鼠神经细胞;三七在保护TX小鼠神经细胞过程中未发挥明显效应。Objective To screen the effective traditional Chinese medicine （TCM） components in compound Gandou Decoction that are responsible for protection of nerve cells in TX mice based on uniform design. Methods A uniform design table with six factors and eleven levels was used in this study. The TX mice were used as the animal model of Wilson＇s disease, and the indices consisted of the protein expression of Bcl-2 and Bax in nerve cells and the content of copper in serum and brain tissue. Multiple stepwise regression analysis was performed to figure out the compatibility of the six effective TCM components in Gandou Decoction, Rheum officinale, Rhizoma Curcumae Longae, Coptis chinensis, Alisma orientalis, Lysimachia christinae, and Radix Notoginseng. Western blot and plasma mass spectrometry were used to determine the apoptosis-related proteins, Bcl-2 and Bax, and the content of copper in serum and tissue, respectively. Results Compared with the control group, the TX mice had significantly lower expression of Bcl-2 in brain tissue （P〈0.05） and serum content of copper （P〈0.05）, as well as significantly higher expression of Bax （P〈0.05） and content of copper in brain tissue （P〈0.05）. The multiple stepwise regres- sion analysis showed that Rhizoma Curcumae Longae and Rheum officinale were influencing factors for the content of copper in serum and brain tissue, respectively; Coptis chinensis and Alisma orientalis were both influencing factors for the expression of Bax and Bcl-2 in brain tissue; Rhizoma Curcumae Longae and Lysimachia christinae were influencing factors for the Bax/Bcl-2 ratio in nerve cells; there was no relationship of Radix Notoginseng with the above indices. Conclusion Rheum officinale, Rhizoma Cureumae Longae, Coptis chinensis, Alisma orientalis, and Lysimachia christinae in Gandou Decoction synergize to protect nerve cells in the TX mice by targeting multiple pathways; Radix Notoginseng has no substantial contribution to protection of nerve cells in TX mice.

关 键 词：均匀设计法 肝豆汤 效应中药 配伍 

分 类 号：R742.4[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]