Li Shu-Hui, Yu Hong-Li, Wu Hao, Zhang Yuan-Bin, Wang Wei
Pharmacology College, Nanjing University of Chinese Medicine Nanjing 210023, China.
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 Processing Nanjing 210023, China.
Zhongguo Zhong Yao Za Zhi. 2020 Jun;45(11):2546-2551. doi: 10.19540/j.cnki.cjcmm.20200328.308.
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.
本研究旨在探讨石灰和甘草炮制法对半夏毒性凝集素蛋白的影响，阐明石灰和甘草炮制半夏的科学解毒内涵。采用蛋白质免疫印迹法半定量分析半夏及制半夏中凝集素的含量。将生品及凝集素分别用甘草汁、石灰水或二者混合溶液浸泡处理，考察不同处理时间对毒性凝集素蛋白含量的影响。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析处理前后溶液及沉淀中凝集素蛋白条带的变化。采用基质辅助激光解吸电离飞行时间质谱技术对处理前后的蛋白质分子量进行定性分析和比较。结果表明，半夏及制半夏中凝集素的含量分别为5.01%和0.04%，表明炮制可显著降低生品中活性凝集素的含量。结果还表明，生药在石灰水中浸泡1天或在甘草汁中浸泡3天后，凝集素含量显著降低，在混合溶液中效果最佳。定性分析表明，经石灰水浸泡处理后，凝集素蛋白分解为小肽段，而经甘草汁浸泡处理后，凝集素蛋白变性沉淀。甘草汁和石灰水炮制后半夏凝集素蛋白的结构被破坏，毒性凝集素蛋白含量显著降低。这是半夏炮制的解毒机制之一。
