| Pollen Typhae is the dry pollen of Typhae angustifolia Linn.,Typha orientalis Presl or the same genus plant.It has effect of hemostasis by resolving stasis.It is mainly used for varieties of bleeding diseases caused by blood stasis as a common medicine in blood circulation.The effect of carbonized Pollen Typhae was enhanced in hemostasis after being carbonized.And carbonized Pollen Typhae has been used for hemostasis in clinical for hundreds of years.In this paper,we studied the material base of Pollen Typhae and Carbonized Pollen Typhae in compound.The paper was divided into four chapters.The first chapter summarized the progression of Pollen Typhae in chemistry,pharmacology,clinical application,and processing history.The second chapter was about on chemical differences through processing.The third chapter studied the hemostatic mechanism in vitro of Pollen Typhae and carbonized Pollen Typhae.The fourth chapter studied the hemostatic mechanism in vivo of Pollen Typhae,carbonized Pollen Typhae and compound.1 Literature reviewWe have reviewed relative ancient and modern documents,collect many research results and discussion,and summarize a review about the historical change of processing mechanism of Pollen Typhae from the ancient documents,the processing and clinical experience,chemical compositions,pharmacological research and clinical applications.2 Chemical differences through processingAs different pollens have similar appearance,it often can not be identified by appearance,and microscopy is useful in identification of Pollen.In the first segment,the identification of Pollen Typhae and carbonized Pollen Typhae was studied.We compared Pollen Typhae and carbonized Pollen Typhae through HPLC、UPLC-MS,and intended to find different compositions between them.We found that,after the processing,the flavonoid changed significantly,in which the flavonoid glycosides were destroied in large extent,while the relative content of flavonoid aglycone increased significantly.Besides,some little molecules compounds with big polarity were produced after carbonized.These changes leaded to pharmacodynamic differences between Pollen Typhae and carbonized ones.By using the chromatography techniques,17 compounds were isolated and elucidated from EtOAc and BuOH extracts as follows:tetracosanoic acid(1),β-sitosterol(2),nonadecanol(3),naringenin(4),vanillic acid(5),nicotinic acid(6),kaempferol(7),isorhamnetin(8),succinic acid(9),quercetin(10),thymine(11),daucosterol(12),uracil(13),typhaneoside(14),stearic acid propanetriol ester(15),isorhamnetin-3-O-neohesperidoside(16),sucrose(17),Compound(1)、Compound(3)、Compound(6)、Compound(11)were obtained from Thpha angustifolia L.for the first time.Reversed high-performance liquid chromatography(RP-HPLC)fingerprint of Pollen Typhae and carbonized Pollen Typhae was eatablished and analyzed,and quantitatively determined typhaneoside,isorhamnetin-3-O-neohesperidoside and isorhamnetin.The method of the HPLC-fingerprint and quantitative analysis is rapid,simple and accurate with a good reproducibility and can be used for the quality control of Pollen Typhae.From before and after processing fingerprint,we found that the content of typhaneoside and isorhamnetin-3-O-neohesperidoside were decreased significantly,while the content of isorhamnetin was increased significantly.Meanwhile,there also have different degree of change in other chromatographic peak in quantity,which leaded to pharmacodynamic differences between Pollen Typhae and carbonized ones.3 Study on anti-oxidative activity and blood coagulationTo study the effects of antioxidation and blood coagulation of Pollen Typhae and carbonized Pollen Typhae,their antioxidant activity was tested by DPPH method and the capability of deoxidating Fe3+.The results show that in the concentration range,both alcohol and water extracts of Pollen Typhae and carbonized Pollen Typhae had significant effects on scavenging DPPH radical and deoxidating Fe3+.The effects of platelet aggregation induced by ADP and thrombin time were investigated.The results indicated that Pollen Typhae and carbonized one showed inhibitory effects on the platelet aggregation and shortened clotting time.In addition,flavonoid compounds also had anti-platelet aggregation and shortened the thrombin time.Pollen Typhae and carbonized ones both showed a certain effect of resolving stasis by inhibiting platelet aggregation significantly,while the potency of raw materials was stronger than the carbonized ones.Pollen Typhae and carbonized ones could shorten TT significantly while the effect of carbonized Pollen Typhae in shortening TT is stronger than the non-carbonized ones.4 The pharmacodynamic differences of Pollen Typhae and carbonized ones in compound4.1 The influences on hemorheology and thrombin activity in different stages of blood coagulationPollen Typhae,carbonized ones and the compound could improve the abnormal blood flow of blood stasis rat by reducing its whole blood viscosity and ESR,increasing deformability of erythrocyte,decreasing the aggregation.Pollen Typhae,carbonized ones and the compound could shorten blood stasis rat’s PT,TT and APTT significantly,in which the procoagulant activity of carbon materials was stronger than the raw materials.Pollen Typhae,carbonized ones and the compound showed the effect of improving blood circulation of blood stasis rat by reducing its FIB,prolonging clotting time in fibrin formation stage significantly,and the potency of crude drug was stronger than the carbonized ones.The content of FIB could impact the whole blood viscosity and plasma viscosity directly.The stasis becomes more intensified with the content of FIB increasing.4.2 The influences on tissue injury of blood stasis rat’s lung and tone characterPollen Typhae,carbonized ones and the compound could ameliorate the lung tissue injury of blood stasis rat,by improving the state of alveolar congestion,thickening,interstitial pulmonary alveolar heavy bleeding,inflammatory cell infiltration,etc.Visible bruising,ecchymosis or petechiae could be seen in stasis patients,according to traditional Chinese medicine theory.The blood stasis rat’s tone became dark and purple,with petechiae appearing on tongue,while there were edema and hemorrhage in tongue lamina propria,all of which were the signs of stasis.The stasis signs of blood stasis rat’s tongue could be improved significantly after administered Pollen Typhae,carbonized ones and the compound.4.3 The influences on SOD,MDA,NO and NOS of blood stasis rat’s lungPollen Typhae,carbonized ones and the compound could raise the activity of SOD,decrease the MDA level and reduce the damage of oxygen free radical matabolism product to adjust the balance of oxidation-antioxidant system.At the same time,the level of NO,T-NOS and cNOS were raised in blood stasis rats and the level of iNOS was reduced.Therefore,Pollen Typhae,carbonized ones and the compound could protect lungs and inhibit the injury of lung. |
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