| Rubiae Radix et Rhizoma is the dry root and rhizome of Rubia Cordifolia L..Agri culture is in anhui,jiangsu,shandong,henan,shanxi and other places.The Rubiae Radix et Rhizoma is bitter in taste,cold in property,has effects of blood-cooling and blood-s tanching,promoting blood circulation.It can be used in blood-heat bleeding or blood st asis induced bleeding,it is especially applicable for hemorrhagic disease caused by bloo d heat with blood stasis.Rubiae Radix et Rhizoma can dredge meridians,remove stasis,and it is often used to treat amenorrhea,bruises,rheumatoid arthritis and blood stasis.Rubiae Radix et Rhizoma is commonly used in clinic to treat blood heat and hemorrha ge syndrome with Cirsii Herba,Imperatae Rhizoma,Platycladi Cacumen,Gardenia Fruct us,etc.The prospect of its clinical application is broad.After being carbonized,its pro perty of cold is weakened,and it is more astringent,mainly used for hemostasis.It can be useed to treat various haemorrhagic diseases such as hematemesis,hemoptysis,bloo d dysentery,urinating blood,uterine bleeding.At present,Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)is often mixed in clinical applications.The prescri ption of Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)is not cle ar.To standardize their rational applications in clinic,we carry on the research of mater ial basis related to the different clinical uses of processed and unprocessed Rubiae Radi x et Rhizoma,providing a scientific basis for different clinical uses of Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized).This thesis was mainly divided into four main parts.The first part was the literature research,the processing evolution,chemical components,pharmacological activities and clinical application were summarized and analyzed respectively by retrieving literature.The second part studied different chemical compositions of Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized),including using HPLC,UPLC-MS analysis chemistry changes of Rubiae Radix et Rhizoma before and after being carbonized and chemical constituents isolated from Rubiae Radix et Rhizoma.The third part was the research of Rubiae Radix et Rhizoma(carbonized)quality standard,including Rubiae Radix et Rhizoma(carbonized)of processing technology research and establish a method to determinate the quantity of quinones in Rubiae Radix et Rhizoma(carbonized).The fourth part studied the influence of Rubiae Radix et Rhizoma compound or carbonized ones combind Aster tataricus to fevered and bleeding rats.The fiveth studied the influence of Rubiae Radix et Rhizoma compound or carbonized ones combind Aster tataricus to blood stasis rats.The concrete research content is as follows:1 Literature reviewThrough consulting relevant and ancient literature and modern research achievement s of scholars both at home and abroad,We have summarized the processing history,ch emical constituents,pharmacological action and clinical application data of Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)in this paper.This provides a b asis of material basic research for Rubiae Radix et Rhizoma's different use in the comp ound.2 Chemical differences through processingWe compared Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)through HPLC?UPLC-MS,and intended to find different compositions between them.W e found that,after the processing,the anthraquinone changed significantly,in which the anthraquinone glycosides were destroied in large extent,while the relative content of anthraquinone aglycone increased significantly.Besides,the content of mollugin was sig nificantly reduced.These changes leaded to pharmacodynamic differences between Rubia e Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized).By using the chromatog raphy techniques,9 compounds were isolated and elucidated from Rubia Cordifolia as f ollows:1-hydroxy-2-methyl anthraquinone(1),alizarin(2),rubiadin(3),purpurin(4),1,3,6-trihydroxy-2-methylanthraquinone(5),purpuro-xanthin(6),mollugin(7),?-sitosterol(8),1,3,6-trihydroxy-2-methylanthraquinone-3-O-?-rhamnosyl-(1?2)-?-glucoside(9).3 The processing technology and the content determination of quinones of Rubiae Radix et Rhizoma(carbonized)At present for content determination of Rubia Cordifolia has a lot of related research,but the study of Rubiae Radix et Rhizoma(carbonized)content determination is less.This paper founded a quantitative UPLC method for the simultaneous determination of purpuro-xanthin,purpurin,rubimaillin and 1,3,6-trihydroxy-2-methylanthraquinone in Rubiae Radix et Rhizoma(carbonized).This method is more rapid and accurate than HPLC method,to establish the quality standard and provide the basis of the technological standardization of Rubiae Radix et Rhizoma(carbonized).In the "Chinese Pharmacopoeia"(2010 version),processing method of Rubiae Radix et Rhizoma(carbonized)is taking piece or section of Rubia Cordifolia and being stir-fri ed to be black-brown-colored superficially and burnt-brown--colored inside,according to carbonizing process.The product is tiny odor,bitter and astringent in taste.However,the content determination of Rubiae Radix et Rhizoma(carbonized)is not to develop re levant regulations,to control the quality of it.Currently,without a universally accepted processing technology standard for Rubiae Radix et Rhizoma(carbonized),which affects it's curative effect in clinical applications.We found that the concentration of 1,3,6-trih ydroxy-2-methylanthraquinone increased obviously during the process of carbonizing.By further study we found that 1,3,6-trihydroxy-2-methylanthraquinone could inhibit the da mage in human umbilical vein endothelial cells,caused by lipopolysaccharide.It had c ertain protective effect on vascular endothelial cells.So we can see that Rubiae Radix e t Rhizoma(carbonized)has higher content of 1,3,6-trihydroxy-2-methylanthraquinone,and reinforce hemostasia and cruor effects.It can be used as an index component to deter mine the best technique in processing Rubiae Radix et Rhizoma(carbonized).A method was developed for the determination of 1,3,6-trihydroxy-2-methylanthraquinone in Rubiae Radix et Rhizoma(carbonized)by HPLC,to provide evidence for quality evaluation of RubiaeRadix et Rhizoma(carbonized).4 The influence of Rubiae Radix et Rhizoma compound or carbonized ones combind Aster tataricus to fevered and bleeding rats4.1 The influence of Rubiae Radix et Rhizoma compound,carbonized ones compound to fevered and bleeding rat's hemorheology and the action of thrombinThe whole blood viscosity of fevered and bleeding rats had different degree of i ncrease,which affected the normal blood circulation.The middle dose of Rubiae Radix et Rhizoma compound and carbonized ones compound could significantly reduce the wh ole-blood viscosity under the rate of different exchange(P<0.05 or P<0.01),this illustrate d that Rubiae Radix et Rhizoma reflected the efficacy of promoting blood circulation to remove blood stasis,improved the blood circulation of the fevered and bleeding rats.Carbonized ones used in compound improved the abnormal blood circulation in the feve red and bleeding rats slightly weaker than the crude drugs,but showed a significant dif ference.It fully embodies the traditional theory of "carbonizing deposit nature",the carb onized ones not only reinforce hemostasia and cruor effects,still retain some original m edicinal effect.The significant increase of plasma viscosity reflected the increase of blood viscosity.The rise in hematocrit illustrated the rise in blood viscosity.Blood-heat and hemorrhag e can concentrate blood and decrease moisture of plasma.As a result,the content of he moglobin in model rats increased relatively.The middle dose group of Rubiae Radix et Rhizoma could only ameliorate the plasma viscosity of the rats with blood-heat and he morrhage syndrome,but it couldn't ameliorate the abnormal rise in hemoglobin and he matocrit significantly.However,The middle dose group of Rubiae Radix et Rhizoma(carbonized)could not only ameliorate the plasma viscosity of the rats with blood-heat and hemorrhage syndrome,but also ameliorate the abnormal rise in hemoglobin and he matocrit to different degrees.Middle,high dose of Rubiae Radix et Rhizoma compound could significantly short en the thrombin time(TT),however,low,middle,high dose of carbonized ones compou nd could significantly shorten(TT),illustrated that they both could influence the forma tion of thrombin clotting time to achieve hemostasis effect.Middle dose of Rubiae Radi x et Rhizoma compound and middle,high dose of carbonized ones compound could sig nificantly shorten activated partial thromboplastin time(APTT),but the effect of carboniz ed ones compound was more obvious.Low dose of Rubiae Radix et Rhizoma compoun d could markedly prolong the prothrombinase time(PT),whereas middle dose of carboniz ed ones compound could significantly shorten PT.It reflected the different effect used i n Ziwanwan and the processing theory of the crude drugs had the effect of blood coold ing and arresting and promoting blood circulation to remove blood stasis,carbonized on es reinforced hemostasia.Rubiae Radix et Rhizoma compound and carbonized ones com pound both were able to increase the content of FIB of the fevered and bleeding model of rats,promote clotting by the fibrinolytic system,the effect of carbonized ones comp ound were better than Rubiae Radix et Rhizoma compound.4.2 The pathological detection of Fevered and Bleeding rat's stomach and lungRubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)in the com pound can both play an important role in the treatment of lung tissue damage by ameli orating the capillary dilatation and congestion in the alveolar septa,fibroplasia,hemorrha ge in pulmonary interstitial,red blood cells infiltration in alveolar spaces,partial inflam matory cells infiltration,destruction of partial alveolar structure among rats with blood h eat and hemorrhage syndromes.And the effect of Rubiae Radix et Rhizoma(carbonize d)in the compound is better than Rubiae Radix et Rhizoma in the compound.Differe nt dose groups of Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)in the compound could ameliorate different aspects of gastric mucosa tissue damage among rats with blood heat and hemorrhage syndromes.By detecting pathological sectio ns,we could observe the phenomenon of pathological tissue bleeding,in order to furthe r clarify the different hemostatic effects between Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)in Ziwanwan.4.3 The influence of Rubiae Radix et Rhizoma compound,carbonized ones compound t o fevered and bleeding rat's platelet systemThe ADP-induced platelet aggregation(P<0.05 or P<0.01)was enhanced in middle a nd high dose of Rubiae Radix et Rhizoma compound,were the same as low,middle an d high dose of carbonized ones compound,which were better than crude ones.Low and middle of Rubiae Radix et Rhizoma compound achieved hemostasis effe ct by reducing the content of 6-keto-PGF1?.It was no significant interaction of the cont ent of TXB2 and reflected that it played the role of promoting blood circulation.Low,middle and high of carbonized ones compound achieved hemostasis effect by reducing t he content of 6-keto-PGF1? and incresing the content of TXB2,which promoted the vas cular contraction and platelet aggregation of fevered and bleeding rats.Also embodied t he traditional processing theory of Rubiae Radix et Rhizoma(carbonized)that it reinf orced hemostasia.5 The influence of Rubiae Radix et Rhizoma compound or carbonized ones combind Aster tataricus to blood stasis rats5.1 The influence of Rubiae Radix et Rhizoma compound,carbonized ones compound to blood stasis rat's hemorheo logy and the action of thrombinCompared with the blank group,the whole-blood viscosity under the rate of different exchange and plasma viscosity of model group had different degree of increase(P<0.05 or P<0.01),could reflect the red blood cells have a certain degree of aggregation and deformability of change,the blood viscosity increased,which illustrated that blood stasis model was successfully established.The Rubiae Radix et Rhizoma compound could significantly reduce the whole-blood viscosity under the rate of different exchange and plasma viscosity of blood stasis rats,showed the Rubiae Radix et Rhizoma used in compound Ziwanwan played a full role of the promoting blood circulation to remove blood stasis,improved the abnormal blood circulation in the blood stasis rats slightly.The carbonized ones compound could significantly shorten the prothrombinase time(PT),thrombin time(TT),activated partial thromboplastin time(APTT),increase the content of FIB(P<0.05 or P<0.01).The Rubiae Radix et Rhizoma compound could significantly prolong the prothrombinase time(PT),middle dose of Rubiae Radix et Rhizoma compound could significantly shorten the thrombin time(TT),activated partial thromboplastin time(APTT),increase the content of FIB(P<0.05 or P<0.01).The main hemostatic mechanism of carbonized ones compound was to stimulate the intrinsic,extrinsic blood coagulation system and fibrinolysis system.The Rubiae Radix et Rhizoma compound embodied two-way regulating functions on hemostasis.This have the certain relationship with the Rubiae Radix et Rhizoma is used for removing stasis and Rubiae Radix et Rhizoma(carbonized)is used for hemostasis.5.2 The pathological detection of blood stasis rat's stomach and lungRubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)in the compound can both play an important role in the treatment of lung tissue damage by ameliorating the capillary dilatation and congestion in the alveolar septa,fibroplasia,hemorrhage in pulmonary interstitial,red blood cells infiltration in alveolar spaces,partial inflammatory cells infiltration,destruction of partial alveolar structure among rats with blood heat and hemorrhage syndromes.Different dose groups of Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)in the compound could ameliorate different aspects of gastric mucosa tissue damage among rats with blood heat and hemorrhage syndromes,but the differences between each other was not very obvious.5.3 The influence of Rubiae Radix et Rhizoma compound,carbonized ones compound to blood stasis rat's platelet systemThe ADP-induced platelet aggregation(P<0.01)was enhanced in Rubiae Radix et Rhizoma compound,The Rubiae Radix et Rhizoma(carbonized)was through a variety of blood coagulation way to develop its hemostatic effect.The Rubiae Radix et Rhizoma compound showed certain effect of blood stasis rat's ADP-induced platelet aggregation,which were weaker than carbonized ones.This reflected Rubiae Radix et Rhizoma had the effect of removing blood stasis and hemostasis.The content of TXB2 of the model group rised sharply,showed the content of TXA2 increased significantly,which could lead to the formation of blood clots,and the decreasing content of 6-keto-PGF1? was in line with the characteristics of blood stasis model.Low,middle and high of Rubiae Radix et Rhizoma compound achieved removing blood stasis effect by reducing the content of TXB2 and incresing the content of 6-keto-PGF1?(P<0.01).Carbonized ones compound could also reduce the content of TXB2 and increse the content of 6-keto-PGF1?,which was weaker than the Rubiae Radix et Rhizoma compound,stated that Rubiae Radix et Rhizoma(carbonized)mainly played its hemostatic effect.Further demonstrates the different hemostatic effects between Rubiae Radix et Rhizoma and Rubiae Radix et Rhizoma(carbonized)used in Ziwanwan. |
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