Wang Houwei, Dou Yanling, Tian Jingzhen, Li Feng, Wang Shijun, Wang Zhenguo
College ofPharmacy, Shandong University of Traditional Chinese Medicine, Jinan 250355, China.
Zhongguo Zhong Yao Za Zhi. 2009 Feb;34(4):438-42.
To research on the substantial foundation of the medical speciality of Chinese traditional medicines from immunogenicity.
Control antigen with hot nature was prepared from the mixture of the aqueous extracts of three Chinese traditional medicines with three typical hot nature of Alpinia officinarum, Cinnamomum cassia and Curculigo orchioides, while that with cold nature prepared with Rheum palmatum, Anemarrhena asphodeloides, Coptis chinensis, and polyclonal antibody was prepared by immunizing rabbit with control antigen. Dot blotting was performed between the polyclonal antibody of control antigen and the aqueous extracts of nine Chinese traditional medicines on a piece of PVDF membrane, and the blotting signals were analyzed by the software of Quantity One.
Blotting signals with hot control antigen of nine Chinese traditional medicines in descending were Zingiber officinale, Aconitum carmichaeli, Eucommia ulmoides, Fraxinus rhynchophylla, Lonicera japonica, Anemarrhena asphodeloides, Coptis chinensis, Rheum palmatum and Phellodendron chinense, which degree of similarity to control antigen in peak value were 57.33%, 43.56 %, 34.16%, 30.2%, 28.81%, 26.53%, 21.68%, 17.62% and 14.85%, respectively. Blotting signals with cold control antigen were Rheum palmatum, Anemarrhena asphodeloides, Coptis chinensis, Phellodendron chinense, Zingiber officinale, Lonicera japonica, Fraxinus rhynchophylla, Eucommia ulmoides and Aconitum carmichaeli in descending, of which degree of similarity to cold control antigen in peak value were 55.22%, 54.23%, 46.72%, 34.08%, 30.3%, 24.48%, 24.33%, 20.35% and 15.17%, respectively. Results of cluster analysis with Wistar's method showed that nine medicines were classified into two groups, one group included Phellodendron chinense, Anemarrhena asphodeloides, Coptis chinensis, Rheum palmatum, another was Zingiber officinale, Aconitum carmichaeli, Eucommia ulmoides, Fraxinus rhynchophylla, Lonicera japonica.
Blotting signals of nine medicines with control antigen regularly varied with the alteration of medicine nature. The more similarity degree of the tested medicine to control antigen was smaller, the more distance of the tested medicine to control antigen was further. Dot immunoblotting was a practical and effective new method in researching the substantial foundation of the medical speciality of Chinese traditional medicines.
从免疫原性方面研究中药药性的物质基础。
选取具有三种典型热性的中药高良姜、肉桂、仙茅的水提取物混合制成热性对照抗原，选取大黄、知母、黄连制成寒性对照抗原，并用对照抗原免疫家兔制备多克隆抗体。将对照抗原的多克隆抗体与9味中药的水提取物在PVDF膜上进行斑点杂交，用Quantity One软件分析杂交信号。
9味中药与热性对照抗原杂交信号强度由高到低依次为生姜、川乌、杜仲、秦皮、金银花、知母、黄连、大黄、黄柏，其峰值与对照抗原的相似度分别为57.33%、43.56%、34.16%、30.2%、28.81%、26.53%、21.68%、17.62%、14.85%。9味中药与寒性对照抗原杂交信号强度由高到低依次为大黄、知母、黄连、黄柏、生姜、金银花、秦皮、杜仲、川乌，其峰值与对照抗原的相似度分别为55.22%、54.23%、46.72%、34.08%、30.3%、24.48%、24.33%、20.35%、15.17%。用Wistar法进行聚类分析结果显示，9味药分为两组，一组为黄柏、知母、黄连、大黄，另一组为生姜、川乌、杜仲、秦皮、金银花。
9味中药与对照抗原的杂交信号随药性的改变呈规律性变化。被测药物与对照抗原的相似度越小，其与对照抗原的距离越远。斑点免疫印迹法是研究中药药性物质基础的一种实用有效的新方法。
