白芍硫磺熏蒸后四物汤对大鼠尿液中内源性代谢产物的影响研究

Effect of Siwu Tang with Paeoniae Radix Alba Fumigated with Sulfur on Endogenous Metabolites in Urine of Rats

目的:观察白芍硫磺熏蒸后四物汤对大鼠尿液中内源性代谢产物的影响。方法:将80只SPF级SD大鼠分为空白组、未经硫磺熏蒸白芍四物汤组、经硫磺熏蒸白芍四物汤组及四药全经硫磺熏蒸四物汤组,每组20只,雌雄各半。含未经硫磺熏蒸白芍四物汤组给予含未经硫磺熏蒸白芍四物汤提取液,含经硫磺熏蒸白芍四物汤组给予含硫磺熏蒸白芍四物汤提取液,四药全经硫磺熏蒸四物汤组给予含经硫磺熏蒸四药的四物汤提取液,空白组给予等量蒸馏水,剂量为34.15 g/kg (生药量),每天给药1次,连续给药14周。收集4组大鼠第6、8、10、12、14周的尿液,进行色谱及质谱分析,将采集到的数据进行多元统计。结果:4组大鼠的尿液样品从第6周开始分离,第8周和第10周时分离最明显,第12周和第14周在得分图上的位置趋于稳定。根据4组在正、负离子模式下的PCA得分图及载荷图,初步鉴定出13个内源性代谢标志物,主要参与了15条代谢通路,其中4条代谢通路的影响值大于0.1,即苯丙氨酸、酪氨酸和色氨酸生物合成通路,二羧酸代谢通路,戊糖和葡萄糖醛酸酯代谢通路和酪氨酸代谢通路。结论:白芍硫磺熏蒸后四物汤造成的肾毒性可能通过对苯丙氨酸、酪氨酸和色氨酸生物合成通路,二羧酸代谢通路,戊糖和葡萄糖醛酸酯代谢通路和酪氨酸代谢通路的影响而产生。

Objective: To observe the effect of Siwu Tang with Paeoniae Radix Alba fumigated with sulfur on endogenous metabolites in urine of rats. Methods:A total of 80 SPF grade SD rats were divided into the blank group,the Siwu Tang group with Paeoniae Radix Alba not fumigated with sulfur,the Siwu tang group with Paeoniae Radix Alba fumigated with sulfur, and the Siwu Tang group with all four medicinals fumigated with sulfur,with 20 rats in each group,half male,half female. The Siwu Tang group with Paeoniae Radix Alba not fumigated with sulfur was given the extract of Siwu Tang with Paeoniae Radix Alba not fumigated with sulfur;the Siwu Tang group with Paeoniae Radix Alba fumigated with sulfur was given the extract of Siwu Tang with Paeoniae Radix Alba fumigated with sulfur;the Siwu Tang group with all four medicinals fumigated with sulfur was given Siwu Tang with all four medicinals fumigated with sulfur;the blank group was given the same amount of distilled water;all groups were given the administration at a dose of 34.15 g/kg(the amount of crude herbs),once a day for 14 consecutive weeks.The urine of rats in the four groups at the 6th,8th,10th,12thand 14thweeks was collected and analyzed by chromatography and mass spectrometry;the collected data were subjected to multivariate statistics.Results:The urine samples of rats in the four groups were separated from the 6thweek,the separation at the 8thand 10thweeks was the most obvious,and the position on the score map tended to be stable at the12thand 14thweeks. According to the PCA score map and load map of the four groups in the positive and negative ion modes,13 endogenous metabolic markers were preliminarily identified,mainly participating in 15 metabolic pathways, of which four metabolic pathways had an impact value greater than 0.1,namely, phenylalanine, tyrosine and tryptophan biosynthesis pathway, dicarboxylic acid metabolism pathway, pentose and glucuronide metabolism pathway and tyrosine metabolism pathway. Conclusion:The nephrotoxicity caused by Siwu Tang with Paeoniae Radix Alba fumigated with sulfur may be caused by the effect on phenylalanine,tyrosine and tryptophan biosynthesis pathway,dicarboxylic acid metabolism pathway,pentose and glucuronate metabolism pathway and tyrosine metabolism pathway.