甘遂醋制前后萜类成分效应部位抗腹水、泻下作用及组成变化研究

目的?探究甘遂醋制前后抗腹水、泻下作用及组成变化。方法?将甘遂用二氯甲烷和95%乙醇依次提取，获得二氯甲烷部位及非二氯甲烷部位。采用癌性腹水小鼠模型，以腹水量、粪便含水量、尿量及肠道水通道蛋白（AQP）1、AQP3的蛋白表达水平为指标，筛选甘遂效应部位，考察甘遂醋制前后效应部位二氯甲烷提取部位的抗腹水及泻下作用变化。同时对二氯甲烷部位及非二氯甲烷部位，以及醋制前后二氯甲烷部位各成分的相对含量变化进行质谱分析。结果?与模型组相比，生甘遂总提物组、二氯甲烷提取部位高剂量组小鼠腹水量和十二指肠、空肠、回肠及结肠AQP1、AQP3蛋白的表达量均显著降低，粪便含水量显著增加，对尿量无影响；非二氯甲烷部位组无明显降低腹水及增加粪便含水量的作用，表明甘遂二氯甲烷部位是其主要效应部位；与模型组相比，醋品二氯甲烷部位高剂量组仍可显著降低癌性腹水小鼠的腹水量，增加粪便含水量，显著降低十二指肠、空肠、回肠中AQP1、AQP3蛋白表达，但对结肠AQP1和AQP3蛋白表达量无显著影响。与生品二氯甲烷部位组相比，醋制后甘遂二氯甲烷部位组小鼠腹水量显著升高，粪便含水量显著降低。此外，甘遂对腹水中癌细胞存活率没有影响，不能促使其凋亡。质谱分析发现，甘遂中萜类成分主要富集于二氯甲烷部位中，且醋制后二氯甲烷部位16个萜类成分中12个成分相对含量减少，4个成分含量增加。结论?甘遂醋制后仍保留“泻水逐饮”功效，但泻下作用缓和，效应的变化可能与所含萜类成分的组成变化有关。 

Study on the Effects of Anti-Ascites and Anti-Diarrhea and Composition of the Effective Parts of Terpenoids from Kansui Radix Before and After Vinegar Processing

OBJECTIVE?To explore the anti-ascites and anti-diarrhea effects of Kansui Radix and vinegar-baked Kansui Radix. METHODS?Dichloromethane and non-dichloromethane fractions were extracted from Kansui Radix with dichloromethane and 95% ethanol in turn. The ascites volume， fecal water content， urine volume， intestinal expressions of AQP1， and AQP3 proteins were measured as indexes to screen the effective fraction and investigate the anti-ascites and anti-diarrhea effects of dichloromethane fraction before and after vinegar-baked. At the same time， the relative content changes of dichloromethane fraction before and after vinegar-baked were analyzed by mass spectrometry. RESULTS?Compared with the model group， the number of ascites and the expressions of AQP1 and AQP3 in duodenum， jejunum， ileum， and colon of mice in the total-extract Kansui Radix group and high-dose dichloromethane fraction group were significantly decreased， while fecal water content was significantly increased， and urine volume was not changed. The non-dichloromethane fraction group did not notably influence the ascites and fecal water content， indicating that the dichloromethane fraction of Kansui Radix was the main effective fraction. Compared with the model group， the high-dose dichloromethane fraction of vinegar-baked Kansui Radix could significantly reduce the ascites volume， increase the fecal water content， and significantly reduce the expression of AQP1 and AQP3 protein in the duodenum， jejunum， and ileum， but had no significant effect on the expressions of AQP1 and AQP3 in the colon. Compared with the group of dichloromethane fraction of raw Kansui Radix， the ascites volume of the vinegar-baked dichloromethane fraction was significantly increased， and the fecal water content was significantly decreased. Besides， Kansui did not affect the survival rate or apoptosis of cancer cells in ascites. Mass spectrometry analysis showed that terpenoids mainly concentrated in dichloromethane fraction， 12 of the 16 terpenoids decreased， and 4 of 16 increased after vinegar processing. CONCLUSION?The "removing water retention by purgation" effect of Kansui Radix was retained after vinegar processing. Vinegar-baked weakened the purgative effect， which may be related to the change of terpenoid composition.