| Purpose: The aim of this study is to study the content of the different polar parts of the Drug-Pair Peony and Licorice,the effect of four polar parts of the Drug-Pair Peony and Licorice on the uterus in mice and its effect on the MDA,SOD,GSH-PX,ROS,NO,PGE2 and PGF2? in dysmenorrhea in model rats,under the guidance of the traditional Chinese medicine theory and the modern scientific methods.And the extraction process of the best polar part was optimized,the tissue distribution was studied,in order to further study the effects of the Drug Pair Peony and Licorice provide scientific,reasonable and necessary reference.Material and method: 1 The contents of paeoniflorin,paeoniflorin,glycyrrhizin,licorice glycyrrhizin and glycyrrhizic acid in the ethyl acetate fraction,ethanol fraction and water fraction were determined by high performance liquid chromatography with wavelength shift method.The changes of five index components in three polarities were compared.2 Uterine smooth muscle in vitro experiments,Untreated mice isolated uterus and oxytocin-induced uterine contractions in vitro mouse model were treated by four polarity fractions of the Drug Pair Peony and Licorice with low,medium and high doses.Observing the effects of different polar parts of the Drug pair Peony and Licorice Decoction on the uterine contraction amplitude and frequency of contractions.3 Injected diethylstilbestrol and oxytocin into rats to get the dysmenorrheal model and to study the treatment effects of different polar fractions and model group;All rats were detected serum MDA?SOD?GSH-PX?ROS?NO by ELISA;Obtained the living uterine tissue homogenized into homogenate,which determined the activity of PGE2?PGF2? by ELISA.4 The extraction rate and the extraction rate of paeoniflorin,paeoniflorin and glycyrrhizin were taken as indexes,ethanol concentration,ratio of material to liquid,extraction time and extraction times as investigation factors,to optimize the extraction process of the best polar part of the Drug pair Peony and Licorice by L9(34)orthogonal test.5 The method of simultaneous determination of paeoniflorin,paeoniflorin,glycyrrhizin and glycyrrhizin in tissues of rats with dysmenorrhea in ethanol fraction of the Drug pair Peony and Licorice was established by high performance liquid phase wavelength shift method.Result: 1 The content of five indicative components respective are8.718 mg.g-1,24.35 mg.g-1,29.49 mg.g-1,18.59 mg.g-1,12.40 mg.g-1 in ethyl acetate fraction;they are 95.84 mg.g-1,161.8mg.g-1,28.89 mg.g-1,8.991 mg.g-1,29.37 mg.g-1in ethanol fraction;they are 14.25 mg.g-1,22.50 mg.g-1,48.44 mg.g-1,0.7242 mg.g-1,19.30 mg.g-1 in water fraction.2 When the Uterine smooth muscle was not treated,four polarity fractions have not an obvious effect on uterine smooth muscle;When the Uterine smooth muscle was treated,except the petroleum ether,the others have some effects on it 3 Compared with the model group,in serum,except the petroleum ether group,the results of other experimental groups were statistically significance;Compared with the model group,in Uterine tissue homogenate,except the petroleum ether group,the results of other experimental groups were statistically significance.4 The optimum extraction process of the Drug Pair Peony and Licorice was 80% ethanol,10 times of alcohol,3 times and 2 hours.5 The calibration curve was linear within the range of 0.1378?g.g-1 ~ 63.65?g.g-1,and the linear range was 0.1378?g.g-1 ~ 66.15?g.g-1 for paeoniflorin concentration,The linear range of glycyrrhizic acid concentration in the range of 0.1112?g.g-1 ~ 26.70?g.g-1 and 0.1530?g.g-1 ~ 73.45?g.g-1 for glycyrrhizic acid was good.Conclusion: 1 The results of content shows that the index component is the largest in the ethanol part,and the ethanol part is the best polar part.2 Ethyl acetate fraction,ethanol fraction and water fraction had some inhibitory effects on contraction of uterine contraction induced by oxytocin in mice,affecting the uterine contraction amplitude and contraction frequency.3The Drug Pair Peony and Licorice decrease the content of MDA?ROS?NO?PGE2?PGF2?,increase the content of SOD?GSH-PX on primary dysmenorrhea model rats.The Drug Pair Peony and Licorice can treat dysmenorrhea rats,and determine the best polar fraction is the ethanol polar fractions.4 The results showed that the method was simple,reasonable and feasible.5 Paeoniflorin,peony lactone glycosides,glycyrrhizin and glycyrrhizin in rat tissues and organs widely distributed,especially in the large intestine,small intestine in the distribution of more concentrated. |
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