Explored The Scientific Connotation Of Efficacy Differences Of Moutan Cortex, Paeoniae Rubra Radix And Paeoniae Alba Radix Based On The Relevance Of “Variety-ITS2 Sequence-component-efficacy”

Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix are used frequently in clincial, are the plant of Ranunculaceae peony. From ancient times to now, there was a clear difference of the efficacy of the 3 medicinal materials in many traditional medical literatures, and the 3 medicinal materials were used to different diseases in clinical.Generally, Moutan Cortex was used to clear liver fire, cool blood and remove blood stasis; Paeoniae Rubra Radix has the effects of eliminating stasis to stop pain and cooling blood; Paeoniae Alba Radix was used to enrich blood and soften the liver, calm the liver and stop the pain. However, many researchers found that Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix contain phenolic acids, terpene glycoside small molecule compounds and polysaccharide macromolecular compounds. Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix contained similar chemical compositions, but they have different clinical efficacy. At present, the material foundation that caused the different efficacy is not clear. Therefore, this issue will revealed the relevance of “variety-ITS sequence-chemical composition- pharmacolo gical activitie” from the genetic differences, component structure differences and different pharmacological activities of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix, provided the scientific evidence for controling medicines quality and their clinical application.Traditional Chinese Medicine is a complex and orderly whole, “Structural component” theory holds that the whole system of Chinese medicine was constituted by three levels.The first level: monomer composition; the second level: the same category monomers constitute the component, and has a stable monomer ratio of monomer within the struture of component; the third level: each component according to the specific structure ratio to constitute the whole of Traditional Chinese Medicine. In this study, the use of traditional chinese medicine fingerprint, cluster analysis and principal component analysis, combined with “structural components” theory to explore the structureal characteristics of internal components in Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix, provided the basic research to the material base of the different efficiency. This paper will study the folloeing aspects.1.The “varieties-ITS2 sequence-composition” relevance of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba RadixFrom the level of plant gene to investigate the relevance of “varieties-ITS2 sequence”. That shown there are 10 variable sites between Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix and the intraspecific genetic distance of Moutan Cortex was less significantly than the genetic distance between species of Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix by detecting ITS2 sequences. Based on ITS2 sequences to construct the NJ tree shown that different regions Moutan Cortex were one class, Paeoniae Rubra Radix and Paeoniae Alba Radix were one class.From chemical constituents to study the association of varieties-ingredients”. thin layer chromatography shown that there were similar categories spots, but the color and size are different in Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix. Constructed the fingerprint of Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix. It found that medicines were divided into three categories, respectively, Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix by cluster analysis. Principal component analysis found that the representative components of Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix were different. 2.Assessment the difference of the effectiveness of Moutan Cortex and Paeoniae Rubra Radix, Paeoniae Alba Radix(1) The efficacy of enrich blood: To establish mouse model of blood deficiency, through detecting WBC, RAC, Hb, HCT, MCHC in peripheral blood and observing the pathological changes of thymus and spleen. The results of study that Paeoniae Rubra Radix group significantly elevated WBC, HCT; Paeoniae Alba Radix significantly elevated WBC,RBC,HCT and Hb. Both Paeoniae Rubra Radix and Paeoniae Alba Radix group can significantly reduce MCHC; Moutan Cortex group has no obvious effect to the indexs in peripheral blood. Paeoniae Rubra Radix and Paeoniae Alba Radix group improved the thymus and spleen of blood deficiency in different level. Moutan Cortex group improved the pathological changes of spleen.(2) The efficacy of heat and cooling blood, bleeding: To establish rats model of blood heat bleeding, through detecting the PT, APTT,TT,FIB in plasma, whole blood viscosity and plasma viscosity, pathological changes in the liver and lung, results showed that Moutan Cortex and Paeoniae Alba Radix group significantly prolonged APTT,TT, reduced FIB; Paeoniae Rubra Radix group significantly shortened PT, prolonged APTT, and improved blood viscosity, plasma viscosity. Moutan Cortex and Paeoniae Rubra Radix group could improve the injury in liver and lung.(3) Study on the effect of promoting blood circulation: the establishment of acute blood stasis and carotid artery thrombosis models, detected four indexs of coagulation, blood rheology, platelet adhesion rate, fibrinolysis system, the expression of inflammatory proteins and other indexs. The study found that the effect of Paeoniae Rubra Radix on four indexs of coagulation and blood rheology was more obvious than Moutan Cortex and Paeoniae Alba Radix group; Moutan Cortex group significantly inhibited platelet adhesion rate. Moutan Cortex and Paeoniae Alba Radix group down regulated the expression of COX2,TLR4,IL-1β, IL-6; Paeoniae Rubra Radix down regulated the expression of COX2,TLR4,IL-1β, and the effect of Moutan Cortex on inflammatory proteins expression was more significant than Paeoniae Rubra Radix and Paeoniae Alba Radix group.Evaluation of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix’s efficacy by different animal models. Paeoniae Rubra Radix and Paeoniae Alba Radix emphasized on enriching blood. However, the effect of Moutan Cortex, Paeoniae Rubra Radix on activating blood circulation was better than Paeoniae Alba Radix, and Paeoniae Rubra Radix was good at coagulation and hemorheology, Moutan Cortex could effectively control the level of inflammatory protein. 3.The analysis of the efficacy material base structure characteristics of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba RadixCombined with “structural components” theory to explore the composition structure characteristics of phenolic acids and terpene glycosides. The results found that there were significant differences in the composition structure of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix. That provied the basic study to cause the different efficacy of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix.Established the model of H2O2 induced HUVECs injury, the content of LDH, SOD, MDA and the expression of MCP-1 and ICAM-1 as indicators to evaluate the effect of phenolic acids, total glucosinolates components and polysaccharides on protecting HUVECs. The results show that total glucosinolates components and polysaccharides had a good protective effect to HUVECs; the phenolic acids in Paeoniae Rubra Radix and Paeoniae Alba Radix had no significant protective effect, however, the phenolic acids in Moutan Cortex could mitigate vascular endothelial cell injury by anti-oxidative stress.In summary, the association of “varieties-ITS2 sequence-composition” in Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix were studied by the biological molecular techniques, TLC and HPLC. Using a variety of animal models to evaluate the efficacy of the three herbs, to explore the material basis cause efficacy differences of Moutan Cortex, Paeoniae Rubra Radix and Paeoniae Alba Radix combined with “structural components” theory.