法半夏炮制过程对半夏中毒蛋白凝集素蛋白的影响  

Effect of Pinelliae Rhizoma processed with lime and licorice on toxic lectin protein

作　　者：李书晖 郁红礼[1,2,3] 吴皓[1,2,3] 张元斌 王卫[1] LI Shu-hui;YU Hong-li;WU Hao;ZHANG Yuan-bin;WANG Wei(Pharmacology College,Nanjing University of Chinese Medicine,Nanjing 210023,China;Jiangsu Key Laboratory of Chinese Medicine Processing,Nanjing 210023,China;Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processings Nanjing 210023,China)

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省中药炮制重点实验室,江苏南京210023 [3]国家教育部中药炮制规范化及标准化工程研究中心,江苏南京210023

出　　处：《中国中药杂志》2020年第11期2546-2551,共6页China Journal of Chinese Materia Medica

基　　金：国家重点研发计划项目(2018YFC1707003);国家自然科学基金项目(81173549,81573605)。

摘　　要：为探究法半夏炮制过程对其毒性成分凝集素蛋白的影响,阐释法半夏炮制解毒的科学内涵,该研究采用Western blot法半定量分析炮制前后半夏中活性凝集素蛋白的含量;以甘草汁、石灰水以及两者混合溶液浸泡的方法分别处理半夏及凝集素蛋白,考察炮制不同时间对毒性蛋白凝集素蛋白含量的影响;采用SDS-PAGE凝胶电泳分析炮制前后溶液及沉淀中凝集素蛋白的条带变化;采用MALDI-TOF技术对炮制前后的蛋白进行相对分子质量定性分析比较。结果显示,生半夏中含凝集素蛋白5.01%,而法半夏中仅含0.04%,表明炮制后活性凝集素含量显著降低;石灰水浸泡1 d、甘草汁浸泡3 d后,生品中凝集素含量显著降低,混合溶液浸泡的影响最大。定性分析发现,使用石灰水浸泡后,凝集素蛋白被分解为小分子的肽段,甘草汁浸泡,可促使凝集素蛋白变性沉淀。法半夏炮制过程中辅料甘草汁和石灰水浸泡均可破坏毒性成分凝集素蛋白结构,显著降低毒性成分凝集素蛋白的含量,此为法半夏炮制解毒机制之一。The present study was aimed to investigate the effect of lime and licorice processing of Pinelliae Rhizoma on its toxic lectin protein and clarify the scientific detoxification connotation of lime and licorice processing of Pinelliae Rhizoma.Western blot was used to semi-quantitatively analyze the contents of lectin in Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum.Raw products and lectin were treated by soaking in licorice juice,lime solution or mixture solution of these two to investigate the different processing time on the content of toxic lectin protein.SDS-PAGE gel electrophoresis was used to analyze the changes of lectin protein bands in the solution and precipitates before and after processing.MALDI-TOF technology was used to qualitatively analyze and compare the protein molecular weight before and after processing.The results showed that the contents of lectin in Pinelliae Rhizoma and Pinelliae Rhizoma Praeparatum were 5.01%and 0.04%respectively,indicating that processing could significantly reduce the content of active lectin in raw products.The results also showed that the content of lectin in raw drugs decreased significantly after soaking in lime solution for one day or in licorice juice for three day,and the effect was greatest in mixture solution.Qualitative analysis showed that after being treated by soaking in lime solution,the lectin protein was decomposed into small peptide segments,while after being treated by soaking in licorice juice,the lectin protein was denatured and precipitated.The structure of lectin protein in Pinelliae Rhizoma was broken after being processed with licorice juice and lime solution,which significantly reduced the content of toxic lectinprotein.This is one of the detoxification mechanisms of Pinelliae Rhizoma processing.

关 键 词：半夏 毒性成分 炮制 凝集素蛋白 

分 类 号：R283[医药卫生—中药学]