桃红四物汤用于原发性痛经的整合效应评价研究

Evaluation study on integration effect of Taohong Siwu Tang in treating primary dysmenorrhea

目的:评价桃红四物汤及各分离部位对热板致小鼠疼痛,醋酸致小鼠扭体反应,小鼠痛经模型及离体子宫收缩的影响,探讨效应部位群的物质基础。方法:采用大孔树脂分离制备各部位样品(TH-1～TH-15);整体动物实验,采用热板致小鼠疼痛的模型,观察样品对小鼠痛阈值的影响;采用醋酸致小鼠扭体反应的模型,观察样品对小鼠扭体反应的影响;采用小鼠痛经模型,观察样品对小鼠扭体反应,子宫匀浆中钙离子和NO的影响。离体器官实验,采用缩宫素诱导的小鼠离体子宫收缩模型,观察样品对离体子宫收缩指标的影响。用HPLC-DAD法对效应部位群进行含量测定。结果:整合生物效应评价表明,桃红四物汤效应部位除整方水煎液(TH-2)和醇沉上清部位(TH-4)外,TH-9,TH-10和TH-11是主要的效应部位群,对其主要效应成分进行分析,化合物类型包括单萜苷类、苯酞类、有机酸类等。结论:桃红四物汤用于痛经的主要效应部位群为TH-9,TH-10和TH-11,是水溶性小分子部位经大孔树脂吸附醇梯度洗脱得到的30%～50%活性部分,与TH-10,TH-11比较,TH-9的效应较强,这可能与其所含的化合物类型和含量密切相关。

Objective： To evaluate the effect of Taohong Siwu Tang and its fractions on hotplate-induced pain in mice, acetic acid-induced writhing response, dysmenon＇heal model and isolated uterine contraction in vitro in mice, and discuss material basis of effect sites. Method： The samples of fractions were prepared by macroporuus adsorptive resins （TH-1-TH-15）. In the whole animal experiment, the hotplate-induced pain mice model was established to observe the effect of the samples on pain threshold in mice ; the acetic acid-induced writhing response mice model was built to observe the effect of the samples on the writhing response in mice; the mice dysmenorrheal model was established to observe the effect of the samples on the writhing response, and calcium ion （ Ca2 ~ ） and nitric oxide （NO） levels in uterine tissue of mice. In the isolated uterus contraction experiment, the oxytocin-induced isolated uterus contraction mice model was established to observe the effect of the samples on the isolated uterus contraction index. HPLC-DAD method was adopted for the content determination of effect sites. Result： According to the evaluation of the integration geological effect, beside TH-2 and TH-4, other three active fractions （TH-9, TH-10 and TH-1 l ） extracted from Taohong Siwu Tang are the main effect sites. Their chemical components were analyzed and identified as monoterpene glycosides, phthalides, organic acids, etc. Conclusion： The effect sites of Taohong Siwu Tang on dysmenorrhea are TH-9, TH-10 and TH-11, which are 30% -50% active fractions obtained from water-soluble small-molecular fractions by gradient elution using ethanol through macroporous absorption resin. Compared with TH-10 and TH-11, TH-9 shows stronger effect, which may be related to the type and content of chemical components it contains.