Study On The Intervention Mechanism Of Compatibility Of American Ginseng And Salvia Miltiorrhiza On Thrombosis

Objective:To study the mechanism and material basis of the intervention of American ginseng Salvia miltiorrhiza on thrombosis.Methods:1 Network pharmacology researchThrough the network pharmacology method,the analysis files were established by means of TCMSp,TTD,DrugBank and UniProt database,the Cytoscape software package was used for data analysis and network construction,and the relationship between pharmacodynamic material basis and target of American ginseng Salvia miltiorrhiza in preventing thrombotic diseases was demonstrated by advanced bubble chart of Omicshare platform.2 Preparation of active componentsThe three active components of ginsenoside,tanshinone and salvianolic acid were prepared by heating reflux extraction method,macroporous resin chromatography separation and purification method,vacuum drying and freeze drying method.The main constituents in each active component were determined by High Performance Liquid Chromatography.3 Animal experimentSeventy healthy male Sprague-Dawley rats were randomly divided into sham operation group(Sham),model group(Model),positive control group(ASA),American ginseng salvia miltiorrhiza decoction group 1(PSD-1),and American ginseng salvia miltiorrhiza decoction group 2(PSD-2),ginseng salvia miltiorrhiza extractive group 1(PSE-1),and American ginseng salvia miltiorrhiza extractive group 2(PSE-2).The sham operation group and the model group were given 0.5%sodium carboxymethylcellulose solution(CMC),the positive control group was given aspirin,the PSD-1 and PSD-2 groups were given to the ginseng salvia miltiorrhiza decoction,and the PSE-1 and PSE-2 groups were given Ginsenoside,tanshinone,and salvianolic acid extract.All the rats were administered by intragastric administration once a day for 7 days.30%FeCl3 filter paper was applied to the carotid artery of rats to induce thrombosis.The ADP-induced platelet aggregation rate was measured by turbidimetry,and the platelet aggregation inhibition rate was calculated.Plasma TXB2,6-K-PGFlα,GPIIb/IIIa,P-sel,β-TG,TNFa,CRP levels were measured by ELISA,and arterial thrombosis was observed by HE staining.Results:1 Based on the network pharmacology method,the active components ofanti-thrombosis of American ginseng salvia miltiorrhiza and its relatedmechanism of action are as follows:The active constituents of the American ginseng Salvia miltiorrhiza can act on 11 targets related to thrombotic diseases and involve 10 signaling pathways.There are 16 active components that can act on the Platelet activation pathway,which are MAPK14,PRKACA-encoded PKA C-alpha,ITGB3-encoded GPIIla,and tanshinones are the most constituents in 16 active components.Among them,tanshinone IIA can act on both target MAPK14 and PRKACA,and cryptotanshinone can act on MAPK14 and ITGB3,beta-sitosterol can also act on target MAPK14 and PRKACA simultaneously.2 The preparation results of the active components of American ginseng salviamiltiorrhiza(1)Detection of medicinal materials:The contents of ginsenoside Rbl,Rgl and Re in the American ginseng herbs were 0.09%,1.18%and 3.47%,respectively,which were 4.74%totally.The contents of cryptotanshinone,tanshinone I and tanshinone IIA in Radix Salviae Miltiorrhizae were 0.12%,0.13%and 0.10%,respectively,totaling 0.36%,and the content of salvianolic acid B was 3.81%.The above measured contents are in compliance with the pharmacopoeia regulations.(2)Content of main components in the effective part:The contents of ginsenoside Rgl,Re and Rbl,which are the main components of ginsenoside extract,were 3.56%,15.07%and 42.64%,respectively,totaling 61.27%.The contents of main components such as dihydrotanshinone,cryptotanshinone,tanshinone I and tanshinone IIA were 2.50%,12.52%,13.90%and 10.61%,respectively,which were 39.52%.The content of salvianolic acid B in salvianolic acid extract was not accurately determined due to the difference in solubility of the sample solution,but under the experimental conditions,the content was about 15%.3 The experimental results of the intervention of the effective parts of Americanginseng salvia miltiorrhiza on rat carotid artery thrombosis are as follows:(1)Platelet aggregation:Compared with the Sham group,the platelet aggregation rate in the Model group was significantly higher(P<0.05);the ASA group,PSD-1 group and PSE-1 group were lower than the Model group.The difference was significant(P<0,05),platelet aggregation inhibition rate was 36.526%,25.260%,19.179%;PSD-2 group,PSE-2 group also decreased compared with Model group,but no significant difference(P>0.05),platelet aggregation inhibition rate was 5.05%and 16.55%,respectively.There was no significant difference in platelet aggregation rate between PSD-1 and PSE-1,PSD-2 and PSE-2.(2)Plasma TXB2,6-K-PGFla content and TXB2/6-K-PGFla ratio:Compared with Sham group,plasma TXB2 level in Model group was significantly increased,there was a significant difference(P<0.07),and the ratio of the two was increased.High,there was significant difference(P<0.05),plasma 6-K-PGFla level decreased,no significant difference;compared with Model group,PSD-1,PSE-1,PSD-2 group plasma TXB2 level decreased,there was significant Sexual differences(P<0.05),plasma TXB2 levels in ASA and PSE-2 groups,the ratio of the two were significantly lower,there was a significant difference(P<0.01);there was no significant difference in plasma 6-K-PGFla level between the groups.Variety.There were no significant differences in plasma TXB2,6-K-PGFla levels and their ratios between PSD-1 and PSE-1 groups,but plasma 6-K-PGFla levels in PSE-2 group were significantly higher than those in PSD-2 group(P<0.05).(3)Plasma GPIIb/IIIa content:Compared with Sham group,plasma GPIIb/IIIa levels in Model rats were significantly increased,the difference was extremely significant(P<0.01),ASA group,PSD-1 group,PSE-1 group,The PSD-2 group and the PSE-2 group were lower than the Model group,and the difference was significant(P<0.05).There was no significant difference in plasma GPIIb/IIIa between PSD-1 and PSE-1,PSD-2 and PSE-2.(4)Plasma β-TG and P-sel content:Compared with Sham group,plasma β-TG and P-sel levels in Model group were significantly increased(P<0.01);compared with Model group.The levels of plasma β-TG and P-sel in ASA group and PSE-1 group were significantly lower(P<0.01),and the levels of P-sel in PSD-2 group and PSE-2 group were significantly lower(P<0.05).The level of β-TG was decreased,but no significant difference was found(P>0.05).The levels of β-TG and P-sel in plasma were decreased in PSD-1 group,and there was significant difference in β-TG(P<0.05),There was no significant difference in plasma P-sel and beta-TG levels between PSD-1 and PSE-1,PSD-2 and PSE-2.(5)Plasma TNFp and CRP levels:Compared with the Sham group,plasma CRP and TNF-α levels in the Model group were significantly higher(P<0.05);compared with the Model group,ASA group,PSD-1 group,Plasma levels of TNF-α were decreased in PSE-1 group,PSD-2 group and PSE-2 group(P<0.01)(P<0.05).Compared with Model group,PSD-1 group,PSE-1 group and PSD-2 group.Plasma CRP levels were decreased in the group and the PSE-2 group,and the difference was significant(P<0.05).There was no significant difference in plasma TNFα and CRP levels between PSD-1 and PSE-1,PSD-2 and PSE-2.(6)Histomorphological changes:The carotid artery of normal rats showed good elasticity and ruddy color by naked eye observation.The carotid artery of model rats showed increased fragility,dark red color and thrombosis.Vascular thrombosis in the carotid artery model segment of the model rats,thrombosis in the lumen was observed under light microscope,the endothelial cells were disordered,the continuity was destroyed,and there were different degrees of separation and exfoliation.There were different degrees of stiffening and straightening of the membrane.The thrombus of each group was dissolved to some extent,and the stiffness of the elastic membrane was reduced.Conclusion:The compatibility of American ginseng and Salvia miltiorrhiza has antithrombotic effect.The ginsenoside,tanshinone and salvianolic acid sites may be the anti-thrombotic pharmacological substance basis,and can play a role through anti-platelet activation and anti-inflammatory.