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目的基于超分子印迹模板自主识别理论,以茜草素为印迹模板功能单体进行猪不同组织的体外亲和色谱实验,验证茜草素归肝经机制,为创立中药归经体外研究方法奠定基础。方法将猪的肝、肺、舌、腹部肌肉、腿部肌肉等组织作固定相制成色谱柱,以PBS为流动相,进行归肝经的茜草素与归脾经的白砂糖(阴性对照)体外亲和色谱实验,采用HPLC测定各洗脱液中茜草素的量,UV法测定阴性对照组各洗脱液中葡萄糖的量,采用总量统计矩分析法获得各亲和色谱参数,获得色谱方程。结果茜草素在肝、肺、舌、腹部肌肉、腿部肌肉的色谱参数为C0:0.250 9~4.813 mg/m L;AUC:2.509~48.13 mg;VR:20.42~30.77 m L;σ2:282.6~532.7 m L2;H:14.13~26.63 m L;n:1.212~1.777;S:0.759 0~1.000。肝脏对茜草素的吸附性最强,腹部肌肉、腿部肌肉、肺、舌对茜草素的吸附作用依次减弱。阴性对照组中,肝脏对葡萄糖的吸附性并非最强。结论猪的肝器官对茜草素的吸附最大,两者存在超分子印迹模板亲合作用,这与茜草归肝经理论相吻合,由此可建立中药归经的体外初步研究方法。
OBJECTIVE Based on the autodifferentiation theory of supramolecular blotting template, the in vitro affinity chromatography experiments of different tissues of pigs with alizarin as the functionalized imprinted template were carried out to verify the mechanism of phletilisin-induced liver metastasis, which laid the foundation for the establishment of the traditional Chinese medicine study in vitro. Methods The liver, lung, tongue, abdominal muscles, leg muscles and other tissues of pigs were used as the stationary phase. The mobile phase consisted of PBS, In vitro affinity chromatography, the amount of rapamycin in each eluate was determined by HPLC, the amount of glucose in each eluate of the negative control was measured by UV method, and the total amount of affinity chromatography was used to obtain each affinity chromatographic parameter to obtain the chromatogram equation. RESULTS: The chromatographic parameters of the radish in the liver, lung, tongue, abdominal muscles and leg muscles were C0: 0.250 9-4.813 mg / m L; AUC: 2.509-48.13 mg; VR: 20.42-30.77 m L; 532.7 m L2; H: 14.13-26.63 m L; n: 1.212-1.777; S: 0.759 0-1.000. The liver adsorption of the strongest Rubidium, abdomen muscles, leg muscles, lungs, tongue adsorption of rubidium in turn weakened. Negative control group, the liver’s adsorption of glucose is not the strongest. Conclusion The liver organ of porcine has the strongest adsorption of raprosyrin, and the two have the affinity of supramolecular imprinting template, which agrees well with the theory of the liver by the Madder, which can establish the preliminary research method of the Chinese medicine.