Study On The Establishment Of The Fingerprint Of Embelia Parviflora Wall. Ex A. DC Of Guangxi National Medicine And Its Effect On Promoting Blood Circulation And Blood Circulation

Objective:1.In order to establish the fingerprint of Guangxi ethnic medicine Embelia parviflora Wall.ex A.DC,provide a basis for the quality evaluation of Embelia parviflora Wall.ex A.DC;2.To study the blood-activating effect of Embelia parviflora Wall.ex A.DC,observe the effect of different polar extracts on blood stasis rats by intragastric administration of Embelia parviflora Wall.ex A.DC;3.To study the blood-suppening effect of Embelia parviflora Wall.ex A.DC.Observe the effect of different polar extracts on blood-deficient mice by intragastric administration of Embelia parviflora Wall.ex A.DC.Method:1.Research on fingerprint:Using high performance liquid chromatography with Phenomenex Luna C₁₈（5μm,4.6 mm×250 mm）column;the mobile phase was eluted with acetonitrile-0.1%phosphoric acid gradient at a flow rate of 0.8 m L/min;the detection wavelength at 225nm and column temperature was 30℃,injection volume is 10μL.2.Study on blood-activating effect:SPF grade SD rats were randomly divided into groups according to body weight:normal control group,model control group,positive control group,Embelia parviflora Wall.ex A.DC petroleum ether site,ethyl acetate group,n-butanol group,Water part group.The test drug was administered by intragastric administration on the first day of the experiment.The doses were as follows:positive group（danshen tablets,dose 0.324 g/kg）,petroleum ether group（4.2 g/kg）,ethyl acetate group（10.64g/kg）,n-butanol group（22.078 g/kg）,Water part group（5.0 g/kg）,blank control group and model control rats were given the same amount of distilled water 2 times a day,continuous administration for 7 days.Day 1 of administration,except the normal group,the other groups of rats were placed in special cages,and then placed in a freezer at（-18±2℃）for continuous freezing twice a day for 2 hours for 7 days.Coagulation syndrome rat model.Throughout the experimental procedure,the room temperature was strictly controlled at 20±2℃,and the animals were fed a normal diet.After 12hours of freezing,rats were anesthetized with 10%chloral hydrate（0.40m L/100g）,and 5 m L of blood was taken from the abdominal aorta.After standing at room temperature for 4 hours,centrifuge at 3500-4000 r/min for10 min at 4℃to dispense serum.Store in a refrigerator at 20℃for use and test the corresponding indicators.3.Study on blood-filling effect:Selection SPF-grade KM female mice were randomly grouped according tobody weight:normal control group,model control group,positive control group,Embelia parviflora Wall.ex A.DC petroleum ether site,ethyl acetate group,n-butanol group,Water part group.The test drug was administered by intragastric administration on the first day of the experiment.The doses were as follows:petroleum ether（5.99g/kg）,ethyl acetate（15.21 g/kg）,n-butanol（31.54 g/kg）,water（7.15g/kg）,the positive control group was given Danggui Buxue Oral Liquid（2.6m L/kg）,and the blank control group and the model control group were given equal volume distilled water once a day,continuous administration for 14 days.On the 12th and 13th days of preventive administration,except for the blank group,each mouse were injected intraperitoneally with cyclophosphamide 80mg/kg for 2 days to prepare a model of blood deficiency in mice.The model of blood-deficiency was continued to be administered during the modeling period,and the blank control group was injected subcutaneously with the same volume of normal saline.30 minutes after the last administration,the eyeballs were taken from the eyeballs,and allowed to stand at room temperature for 4 hours.After centrifugation at 3500-4000r/min for 10minutes at 4,the serum was dispensed and stored in a refrigerator at-20℃for detection of the corresponding indicators.Result:1.Established the common pattern of Guangxi national medicine Embelia parviflora Wall.ex A.DC Fingerprint map.And identify the components of two characteristic peaks（catechin and gallic acid）.2.SPSS cluster analysis was carried out to classify Embelia parviflora Wall.ex A.DC from different origins into two categories,three of which were clustered into one category,and the other producing areas were clustered into one category.3.The similarity between HPLC fingerprint and control map of 12batches of Embelia parviflora Wall.ex A.DC was>0.94,indicating that the chemical composition of the medicinal materials of each producing area had good consistency,indicating that there was not much difference between the different origins of Embelia parviflora Wall.ex A.DC,but in terms of content There will be some differences.4.According to the results of principal component analysis,samples of Embelia parviflora Wall.ex A.DC from different producing areas can be divided into two categories:Embelia parviflora Wall.ex A.DC in three producing areas,and other origins in another.5.Compared with the control group,the serum levels of TXB₂,TXB₂/6-keto-PGF_1α,PAI-1 and c AMP were increased（P<0.01）,6-keto-PGF_1αcontent,The t-PA content and t-PA/PAI-1 ratio decreased（P<0.01）.6.Compared with the model control group,the different polar extracts of Embelia parviflora Wall.ex A.DC could reduce the content of TXB₂,PAI-1and c AMP in the serum of rats,and increase the content of 6-keto-PGF_1αand t-PA,especially The effects of the ethyl acetate group and the n-butanol group were significant,and the difference was statistically significant.7.The administration of cyclophosphamide resulted in a corresponding decrease in the number of white blood cells,red blood cell count,hemoglobin content,and platelet count in the peripheral blood of mice,indicating that the blood deficiency model was successfully modeled.8.Compared with the model control mice,the administration group of Embelia parviflora Wall.ex A.DC extract,especially the n-butanol group and the water group,can significantly increase the number of red blood cells,white blood cells,hemoglobin,and red blood cells in the peripheral blood of mice with blood deficiency caused by cyclophosphamide.The specific gravity level and platelet count were statistically significant.9.The administration groups of Embelia parviflora Wall.ex A.DC extract,especially n-butanol group and water part group,can improve the factors related to hematopoietic function such as IL-3,EPO,G-CSF,M-CSF and VCAM-1 in blood deficiency mice.The difference was statistically significant.10.The administration groups of Embelia parviflora Wall.ex A.DC extract,especially the n-butanol group and the water part group,can increase the levels of IL-2,IL-6 and other immune-related factors in blood-deficient mice,and reduce the spleen index of blood-deficiency mice.The thymus index of blood-deficient mice was increased,and the difference was statistically significant.Conclusion:1.The established HPLC fingerprint method is simple in operation,stable and efficient,and has strong specificity,which can provide reference for further research on the quality evaluation of Embelia parviflora Wall.ex A.DC and the basic research of pharmacodynamic substances.2.Different polar extracts of Embelia parviflora Wall.ex A.DC can significantly increase the number of peripheral blood red blood cells and platelets in mice with blood deficiency caused by cyclophosphamide,increase the content of hemoglobin,and increase the hematocrit level.3.Different polar extracts of Embelia parviflora Wall.ex A.DC can increase the expression levels of cytokines related to hematopoietic function such as IL-2,IL-3,IL-6,EPO,G-CSF,M-CSF and VCMA-1,indicating that Angelica sinensis has blood-supplying effect.4.Different polar extracts of Embelia parviflora Wall.ex A.DC can increase the content of 6-keto-PGF_1αin serum,decrease the content of TXB₂and c AMP in serum.And increase the content of t-PA,decrease the content of PAI-1,and maintain the balance of TXB₂/6-keto-PGF_1αin blood.Adjusting the balance of t-PA/PAI-1 and correcting the effect of its imbalance,indicating that Embelia parviflora Wall.ex A.DC has the effect of promoting blood circulation.