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ObjectiveTo establish blood stasis models in zebrafish using three inducers and select the optimal model for evaluating the activity of Notoginseng Radix et Rhizoma in promoting blood circulation. MethodArachidonic acid （AA）， ponatinib， and isoprenaline （ISO） were used to induce blood stasis models in zebrafish. A normal group， a model group， a positive drug group， and Notoginseng Radix et Rhizoma water extract freeze-dried powder groups at different concentrations were set up. The staining intensity of cardiac erythrocytes and the fluorescence intensity of cardiac apoptotic cells were calculated， the anti-thrombotic effect and anti-myocardial hypoxia activity of Notoginseng Radix et Rhizoma were evaluated. The activities of water extract and 70% methanol extract of Notoginseng Radix et Rhizoma were compared based on the preferred AA- and ISO-induced blood stasis models in zebrafish and the difference in the chemical composition was analyzed by UHPLC LTQ-Orbitrap MS/MS. ResultAfter induction by AA and ponatinib， the staining intensity of cardiac erythrocytes was reduced （P<0.01）， and the fluorescence intensity of cardiac apoptotic cells increased after the induction by ISO （P<0.01）. The freeze-dried powder of the water extract of Notoginseng Radix et Rhizoma could antagonize the thrombosis in the AA-induced model （P<0.01） and the myocardial apoptosis in the ISO-induced model （P<0.05）， while no significant improvement in the thrombosis was observed in the ponatinib-induced model. The freeze-dried powder of 70% methanol extract of Notoginseng Radix et Rhizoma could inhibit myocardial apoptosis in the ISO-induced blood stasis model （P<0.01）， and the effect was stronger than that of the freeze-dried powder of Notoginseng Radix et Rhizoma water extract. The difference in chemical composition lay in some saponins （such as ginsenoside Re）， amino acids， and acetylenic alcohols. ConclusionAA， ponatinib， and ISO all can serve as inducers for the blood stasis model in zebrafish. AA- and ISO-induced models can be used to evaluate the activity of freeze-dried powder of Notoginseng Radix et Rhizoma water extract in promoting blood circulation. The chemical compositions of the freeze-dried powders of Notoginseng Radix et Rhizoma extracted with water and 70% methanol are quite different. For the ISO-induced blood stasis model， the freeze-dried powder of Notoginseng Radix et Rhizoma extracted with 70% methanol has a stronger ability against myocardial hypoxia. Saponins and acetylenic alcohols may be closely related to the effects of promoting blood circulation and resolving blood stasis.
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Objective: To elucidate the rationality of integrated processing technology of Leonuri Herba based on comparison of chemical constituents and pharmacological effect of Leonuri Herba between traditional and integrated processing technology. Method:The contents of stachydrine hydrochloride,leonurine hydrochloride,rutin,hyperoside and isoquercetin were used as indexes to compare the differences in the contents of chemical constituents between traditional and integration processing technology of Leonuri Herba.Effect of Leonuri Herba with different processing technology on auricular swelling induced by dimethylbenzene in mice were observed to compared the differences of their anti-inflammatory effect.And rat acute blood stasis model was used to compare the differences of Leonuri Herba with different processing technology on hemorheology and blood coagulation indexes. Result:Contents of stachydrine hydrochloride,leonurine hydrochloride,rutin,hyperoside and isoquercetin in products of integrated processing were 1.558%,0.168%,0.137%,0.113% and 0.078%,they were 1.482%,0.134%,0.125%,0.082% and 0.071% in products of traditional processing,respectively.Both of the processing methods could reduce the degree of swelling and the contents of tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-6 in mouse serum.And both of the processing methods could reduce the whole blood viscosity,prolong the thrombin time(TT),prothrombin time(PT),activated partial thromboplastin time(APTT) and reduce the concentration of plasma fibrinogen(FIB) in acute blood stasis model rats. Conclusion:Compared with the traditional processing technology,the integrated processing technology is better in guaranteeing the quality of Leonuri Herba decoction pieces and reducing the production cost,which indicates that the integrated processing of Leonuri Herba is reasonable.
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OBJECTIVE： To study the improvement effects of fibrinolytic enzyme from Sipunculus nudus （SNFE） on hemorheology disorder and vascular endothelium injury in naked acute blood stasis model rats. METHODS： SD rats were randomly divided into control group， model group， aspirin group （100 mg/kg） and SNFE high-dose and low-dose groups （2 500， 5 000 U/kg）， with 10 rats in each group. They were given relevant medicine intragastrically once a day， for consecutive 7 d. One hour after the 6th day of administration， except for control group， other groups were given adrenaline hydrochloride 0.8 mg/kg subcutaneously， and then the acute blood stasis model was induced by ice-water bath. Blood was collected from abdominal aorta 2 h after the next day. Blood rheological parameters such as whole blood viscosity （high， medium and low shear rate）， plasma viscosity， hematocrit， erythrocyte aggregation index and erythrocyte deformability index were measured by automatic rheometer. The contents of NO and ET-1 in plasma and their ratio were determined by ELISA， and the damaged degree of vascular endothelium were observed by HE staining. RESULTS： Compared with control group， whole blood viscosity of high， medium and low-shear rate， plasma viscosity， erythrocyte aggregation index and ET-1 content were increased significantly in model group， while erythrocyte deformability index， NO content and NO/ET-1 ratio were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Compared with model group， whole blood viscosity of high， medium and low-shear rate， plasma viscosity， hematocrit， erythrocyte aggregation index and ET-1 content were decreased significantly in SNFE high-dose groups. Erythrocyte deformability index， NO content and NO/ET-1 ratio were increased significantly， with statistical significance （P＜0.05 or P＜0.01）. In SNFE low-dose group， erythrocyte deformability index and NO/ET-1 ratio were increased significantly， while ET-1 content was decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. Vascular endothelial staining showed that compared with control group， the structure of aorta layers in model group was loose and disordered， the endothelial defect was incomplete， the vacuoles increased， and the endothelial damage was obvious. The endothelium of rats in each administration group was damaged to varying degrees， but the degree of injury was lighter than in model group. CONCLUSIONS： SNFE can improve hemorheological abnormalities and vascular endothelial injury in rats with acute blood stasis.
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OBJECTIVE:To observe the effects of total organic acid of Thladiantha dubia fruit(TOATF)on coagulation time and hemorheology of rats with cold coagulation and blood stasis. METHODS:60 rats were randomly divided in normal control group (water),model control group (water),aspirin group (positive drug,50 mg/kg) and TOATF low-dose,medium-dose and high-dose groups(50,100,200 mg/kg)with 10 rats in each group. Except for normal control group,cold coagulation and blood stasis model was induced by 4 ℃ water bath and subcutaneous injection of adrenaline hydrochloride,and then given correspon-dence medicine intragastrically,once a day,for consecutive 14 d. 24 h after last administration,blood samples were collected from aorta abdominalis. The coagulating time(CT),erythrocrit(HCT),plasma viscosity(PV),prothrombin time(PT),thrombin time (TT),activated partial thromboplastin time (APTT),platelet aggregation rate (PAR) and whole blood low-shear,middle-shear and high-shear viscosity were measured and blood sedimentation equation K value was calculated. RESULTS:Compared with nor-mal control group,CT,PT,TT and APTT of model control group were shortened,and HCT,PV,blood sedimentation equation K value,PAR and whole blood low-shear,middle-shear and high-shear viscosity increased(P<0.05 or P<0.01). Compared with model control group,CT,PT and APTT prolonged in aspirin group and TOATF medium-dose and high-dose groups,and PAR de-creased;PT of treatment groups prolonged,while HCT,PV,blood sedimentation equation K value and whole blood low-shear and middle-shear viscosity decreased(P<0.05 or P<0.01). CONCLUSIONS:TOATF has obvious improvement effects on anticoagula-tion and hemorheology in rats with cold coagulation and blood stasis.
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Objective: To observe the effect of different extracts from carbonized Rubiae Radix et Rhizoma (RRR) on acute blood stasis model of rats, in order to screen the optimal extraction method for its hemostatic effect by removing blood stasis. Methods: The acute blood stasis models of rats were established with sc injection of high-dose adrenaline hydrochloride after continuous administration for 8 d and being socked in ice-water, using Yunnan Baiyao as positive drug. The effect of the alcohol extract, compound (alcohol + aqueous) extract, aqueous extract, and aqueous extract after alcohol extracting on hemorheology, in vitro thrombus, blood plotelets system, and fibrinolytic system in acute blood stasis model of rats were observed. Results: Compared with the control group, the whole blood viscosity in each shear rate (P < 0.01) and the wet and dry weight of in vitro thrombus could be increased (P < 0.05), the coagulation time was prolonged (P < 0.05, 0.01), the FIB content and platelet aggregation rate induced by ADP increased (P < 0.01), the PAI-1 and TXB2 contents increased, and the 6-keto-PGF1α content decreased in the rats of model group (P < 0.05, 0.01). Compared with the model group, both the alcohol extract and compound extract could significantly improve the whole blood viscosity, decrease the wet and dry weight of external thrombus, shorten the coagulation time, increase the contents of PAI-1 and 6-keto-PGF1α, and decrease the TXB2 content (P < 0.05, 0.01). Both of them could make platelet aggregation rate increased, but have no significant influence on the content of FIB. The alcohol extract significantly decreased the content of t-PA (P < 0.05), while the compound extract significantly increased t-PA content (P < 0.01). The compound extract has influenced the hemorheology and the wet and dry weight of in vitro thrombus greater than the alcohol extract. While on the coagulation time, the effect of alcohol extract is greater. Conclusion: The hemostatic effects by removing blood stasis of alcohol extract and compound extract are stronger than those of others, especially the compound extract. In other words, the optimal extraction method of its hemostatic effect by removing blood stasis is the compound extract.
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Objective: To investigate the effects of Moutan Cortex, Paeoniae Rubra Radix, and Paeoniae Alba Radix on the acute blood stasis model rats, and reveal their mechanism and differences in efficacy. Methods: The actue blood stasis model of rats was induced by sc injecting adrenaline hydrochloride and soaking in ice water. Salvia Injection was used as the positive control drug. The changes of viscosity of whole blood and plasma, thrombin activity, blood platelet system, and the contents of TXB2 and 6-keto-PGF1α were detected in the acute blood stasis model rats. Results: Compared with the model group, Moutan Cortex obviously extended PT, and TT (P < 0.05), inhibited platelet adhesion rate (P < 0.01), reduced TXB2 (P < 0.01), increased 6-keto-PGF1α in serum (P < 0.01), and regulated eNOS and p-eNOS expression (P < 0.01, 0.001). Paeoniae Rubra Radix significantly improved whole blood viscosity at different shear rates, extended APTT, TT, and PT (P < 0.05, 0.01), reduced platelet adhesion rate (P < 0.05), decreased TXB2, increased 6-keto-PGF1α in serum (P < 0.01), and increased eNOS and p-eNOS expression (P < 0.05). Paeoniae Alba Radix had no significant effect on hemorheology, but could significantly prolong APTT and TT (P < 0.05), reduce platelet adhesion rate (P < 0.05), increase the content of 6-keto-PGF1α in serum (P < 0.05) and increase the eNOS and p-eNOS expression in platelet. Conclusion: All Moutan Cortex, Paeoniae Rubra Radix, and Paeoniae Alba Radix have varying degrees on hemorheology, coagulation system, platelet system, but Moutan Cortex and Paeoniae Rubra Radix have the better role in promoting blood circulation compared with Paeoniae Alba Radix.
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OBJECTIVE: To observe the effect of Shuxuening injection (traditional Chinese medicines) on the auricle microcirculation in mice. METHODS: The auricle microcirculation was closely observed to examine the effects of Shuxuening injection on the auricle arteriole and venule diameters, number of open capillary cross-network and blood flow in the normal mice and acute blood stasis mice. RESULTS: Shuxuening injection significantly increased the diameters of arterioles and venule in normal and acute blood stasis model mice, increased the number of open capillaries and improved the flow of blood stasis mice. CONCLUSION: Shuxuening injection can significantly improve the auricles microcirculation in mice.
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OBJECTIVE:To observe the influence of total Saponin of Frucius Tribuli on hemorheological and thrombosis in vitro in model rats with blood stasis.METHODS:The model rats were randomly divided into control,model,ligustrazine and total saponin of Frucius Tribuli(high,medium,and low dosage)group,all administered drugs through stomach irriga?tion.Except the control group,blood stasis models were established in the rest five groups,and hemorheological parameters and thrombosis indices were determined in these groups.RESULTS:In the model group,blood viscosity increased;red blood cell electrophoresis period was prolonged;fibrinogen content,thrombus length and weight increased too.However,blood viscosity,red blood eletrophoresis period,fibrinogen content as well as the wet and dry weight and length of thrombus in mice of total saponin group decreased significantly,as compared with the model group(P
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Objective: To observe the main pharmacodynamic effects of Yinaoning (YNN) on acute blood stasis rats and cerebral ischemia rats. Methods: Rats were prevented by the oral administration of YNN (0.4375～ 1.75g/kg once daily for 7 days. The acute blood stasis model and cerebral ischemaed model were used. Results: YNN could decrease the blood viscosity and plasma viscosity, shorten the length of in vitro thrombus, abate the wet and dry weights of thrombus in the acute blood stasis model group. It decreased the brain index significantly in the cerebral ischemia model group. The high dose of YNN could decrease the level of Evans blue obviously and reduce the degeneration of cranial nerve cells. These effects were similar to those of YNN Tablets and were dose dependent. Conclusion: YNN is effective for acute blood stasis rats and cerebral ischemaed rats.
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AIM: To observe the ameliorative effect of Puerarin Injection on blood hyperviscosity in acute blood-stasis model rats. METHODS: The acute blood-stasis model rats were made by injected adrenaline repeatedly and soaked in ice water. The markers of whole blood viscosity and plasm viscosity in the rats were measured with Auto-Viscometer. RESULTS: The whole blood viscosity and plasm viscosity was significantly raised in the acute blood-stasis model rats and Puerarin Injection had ameliorative effects on them. CONCLUSION: Puerarin Injection can ameliorate the blood-high-viscosity in rats of blood-stasis model.