Theoretical exploring of potential mechanisms of antithrombotic ingredients in danshen-chishao herb-pair by network pharmacological study, molecular docking and zebrafish models.

BACKGROUND: Salvia miltiorrhiza (Danshen, DS) and Radix Paeoniae Rubra (Chishao, CS) herbal pair (DS-CS) is a famous traditional Chinese combination which has been used as antithrombotic formular for centuries. However, there is still lack of sufficient scientific evidence to illustrate its underlying mechanisms. The purpose of this study is to investigate the antithrombotic effects of DS-CS extract in zebrafish and explore its possible mechanism of action. METHODS: The quality of traditional Chinese medicines DS and CS granules was evaluated using High Performance Liquid Chromatography (HPLC). Subsequently, the therapeutic effect of the DS-CS combination and its components, Salvianolic Acid A (SAA) and Paeoniflorin (PF), in various concentrations on thrombosis was experimentally validated. Moreover, the interaction between DS-CS and the thrombosis disease targets was analyzed through network pharmacology, predicting the potential antithrombotic mechanism of DS-CS. Molecular docking and in vivo zebrafish experiments were conducted to validate the predicted targets, with qRT-PCR utilized for target validation. RESULTS: DS-CS exhibited anti-thrombotic effect in zebrafish with concentrations ranging from 25 to 300 µg/mL. The co-administration of PF and SAA at 25 µg/mL each revealed a synergistic antithrombotic effect exceeding that of individual components when contrasted with PHZ treatment. Protein-protein interaction (PPI) analysis identified key genes, including Albumin (ALB), Proto-oncogene tyro-sine-protein kinase Src (SRC), Matrix metalloproteinase-9 (MMP9), Caspase-3 (CASP3), Epidermal growth factor receptor (EGFR), Fibroblast growth factor 2 (FGF2), Vascular endothelial growth factor receptor 2 (KDR), Matrix metalloprotein-ase-2(MMP2), Thrombin (F2), and Coagulation factor Xa (F10), associated with the antithrombotic action of PF and SAA. Furthermore, KEGG pathway analysis indicated involvement of lipid metabolism and atherosclerosis pathways. Molecular docking revealed strong binding of PF and SAA to pivotal hub genes, such as SRC, EGFR, and F10. The experimental findings demonstrated that DS-CS could upregulate the mRNA expression levels of EGFR while inhibiting F10 and SRC mRNA levels, thereby ameliorating thrombotic conditions. CONCLUSION: This research provided valuable insights into the potential mechanisms underlying the antithrombotic activity of DS-CS. Our findings suggested that PF and SAA could be the key active ingredients responsible for this activity. The antithrombotic effects of DS-CS appeared to be mediated through the regulation of mRNA expression of SRC, EGFR, and F10. These results enhanced our understanding of DS-CS's therapeutic potential and lay the groundwork for future studies to further elucidate its mechanisms of action.

Antiplatelet and Antithrombotic Properties of Compound L-36, a 6H-1,3,4-Thiadiazine Derivative.

Compound L-36, a new derivative of 6H-1,3,4-thiadiazine, was studied in in vitro and in vivo experiments. This compound exhibits high antiplatelet and antithrombogenic activity. In in vitro experiments, compound L-36 by its antiplatelet activity (by IC50) was superior to acetylsalicylic acid by 9.4 times. In in vivo experiments, compound L-36 by its ED50 value was close to the comparison drug. On the model of pulmonary artery thrombosis, compound L-36 ensured better survival of experimental animals than acetylsalicylic acid. Morphological studies showed that compound L-36 effectively attenuated the thrombosis processes in the pulmonary tissue induced by intravenous injection of a thrombogenic mixture (epinephrine and collagen).

Screening and validation of antithrombotic effective components group from Trichosanthes extract / 药学学报

To screen the antithrombotic effective components group of Trichosanthes extract, and to verify its pharmacodynamics and analyze its mechanism, the HPLC fingerprint of Trichosanthes extract (0.09, 0.45, 0.9 g·kg-1) was established, and the pharmacodynamic indexes of antithrombosis in rats with aspirin (0.01 g·kg-1) as positive control group were determined (the animals used in this experiment were approved by the Medical Ethics Committee of Wannan Medical College). The antithrombotic spectrum-activity relationship of Trichosanthes extract was studied and the effective antithrombotic ingredients group was screened by grey relational analysis. The monomer compound mixed solution (0.006, 0.03, 0.06 g·kg-1) was prepared according to the content of each component in the active component group, and the pharmacodynamics and action mechanism were studied to verify the correctness of the spectrum-effect relationship. The correlation between the 22 components of Trichosanthes extract and antithrombotic efficacy was different and showed dose-effect relationship. Cytosine, uracil, guanine, hypoxanthine, xanthine, adenine, guanosine, and adenosine are the main antithrombotic components of Trichosanthes extract. The ratio of cytosine, uracil, guanine, hypoxanthine, xanthine, adenine, guanosine and adenosine was 3∶12∶10∶5∶2∶8∶13∶14. Compared with the model group, the thrombus dry weight of each effective components group could be effectively reduced (P<0.01 or P<0.05), but there was no significant difference between each effective components group and the Trichosanthes extract group. Compared with the model group, the TXB2 content in group (0.06 g·kg-1, 0.03 g·kg-1) could be effectively reduced (P<0.01 or P<0.05), and the content of 6-keto-PGF1α could be increased in each group (P<0.01), and the TXB2/6-keto-PGF1α tended to be normal and showed a dose-effect relationship. The effect was better than that in the Trichosanthes extract group (0.45 g·kg-1) (P<0.01). The effective ingredients group has a good antithrombotic effect, its mechanism is to inhibit platelet aggregation and improve vascular endothelial function.

Influence of ceftriaxone on pharmacokinetics, anticoagulation andantithrombotic effects of phenolic acids from Danhong injection in rats / 药学学报

Danhong injection (DHI) and ceftriaxone sodium were used in combination based on their experimental uses in clinic. This study was designed to investigate the impact of ceftriaxone on pharmacokinetics and pharmacodynamics of the phenolic acids from DHI. After administration of DHI for 7 d, ceftriaxone (CFTX) was combined with DHI for the next 7 d in adult male Sprague-Dawley (SD) rats. All the drugs were administered through caudal vein. UHPLC-TQ-MS was applied in determining the plasma concentration of p-coumaric acid (p-CA), salvianolic acid D (SaD), rosmarinic acid (RA) and salvianolic acid B (SaB). The pharmacokinetic parameters of the combination group or the Danhong injection alone group were calculated by statistical moment method, Cmax and the average of the area under the curve AUC0-t using 90% confidence interval of the bioequivalence and bioavailability degree module in DAS 3.2.8 statistic software. The results showed that Cmax of p-CA, SaD, RA and SaB were unqualified within 90% confidence intervals for bioequivalence statistics. And the results showed that AUC0-t of SaD, RA and SaB within 90% confidence intervals for bioequivalence statistics were unqualified. There were no significant difference in the tmax (P>0.05). The results of anticoagulation in vivo showed that the international normalized ratio (INR), prothrombin time (PT), thrombin time (TT) and activated partial thromboplastin time (APTT) were significantly increased when combined with CFTX (P<0.05 or P<0.01). The results in antithrombotic effects revealed that the thromboxane B2 (TXB2) level in serum was significantly decreased (P<0.01) in the combination group compared with Danhong injection alone. However, there was no significant difference in antiplatelet effects. These results suggest that CFTX may enhance the anticoagulation and antithrombotic effects of DHI through altering pharmacokinetics and pharmacodynamics in SD rats.

Antithrombotic effects of cilostazol combined with clopidogrel and aspirin in senile patients with cerebrovas-cular disease after PCI surgery / 中国基层医药

Objective To analyze the antithrombotic effects of cilostazol combined with aspirin and clopi-dogrel in elderly patients with cerebrovascular disease after PCI .Methods 100 elderly patients with cerebrovascular diseases who treated with coronary artery interventional therapy ( PCI) were randomly divided into the control group and the observation group according to the digital table ,50casess in each group.The two groups were given control of blood pressure ,blood lipids ,blood sugar ,improve circulation and other conventional treatment .The control group was treated with aspirin combined with clopidogrel ,the observation group was treated with cilostazol based on the treatment of control group.Before and after treatment for 1,4 and 8 weeks,the platelet aggregation degree was detected by PL-11 automatic platelet analyzer .During 2 months of follow-up,the degree of platelet aggregation ,the volume of platelets,the efficacy of treatment and the incidence of adverse reactions were compared .Results The platelet aggre-gation rate between the two groups had no statistically significant difference before treatment (t0 =2.782,P>0.05). After treatment,the platelet aggregation rate of the two groups decreased significantly ,but after treatment for 1,4 and 8 weeks,the platelet aggregation rates of the observation group were significantly lower than those of the control group [(51.87 ±9.65)％,(40.85 ±10.24)％,(38.52 ±9.64)％;(69.25 ±8.41)％,(62.43 ±9.22)％,(58.46 ± 10.18)％],the differences were statistically significant (t1 =5.693,t4 =4.846,t8 =6.719,all P<0.05).Before treatment,the mean platelet volume between the two groups had statistically significant difference ( t0 =2.146,P>0.05).After treatment,the platelet volume of the two groups decreased significantly ( t1 =1.656,t4 =1.438,t8 =2.189,all P<0.05).There were no statistically significant differences between the observation group and the control group (t1 =3.716,t4 =1.271,t8 =2.523,all P>0.05).The effective rate of the observation group was 94.00％(47/50),which was significantly higher than that of the control group [82.00％(41/50)],the difference was statis-tically significant (χ2 =4.683,P<0.05).The incidence rates of adverse reactions in the observation group and the control group were 10.00％(5/50) and 8.00％(4/50),respectively,there was no statistically significant difference between the two groups (χ2 =1.947,P=0.136).Conclusion Cilostazol combined with clopidogrel and aspirin in the treatment of elderly patients with cerebrovascular disease after PCI can significantly reduce platelet aggregation rate,improve clinical curative effect ,and has certain clinical value .

Biological effects of fucoidan isolated from Fucus vesiculosus on thrombosis and vascular cells

BACKGROUND: Fucoidan is a highly sulfated glycosaminoglycan, which has a molecular structure similar to that of heparin. The antithrombotic effects of fucoidan in vitro have been widely reported, but its antithrombotic effects in vivo as well as its other biological properties in vitro have not been well investigated. METHODS: This study investigated the effects and mechanism of fucoidan from Fucus vesiculosus on thrombosis both in vitro and in vivo. A ferric chloride-induced mouse carotid artery thrombosis model was used to determine the antithrombotic effects of fucoidan in vivo. Additionally, changes in the levels of proinflammatory cytokines and chemokines were examined in vascular cells treated with fucoidan. RESULTS: In vivo studies employing a ferric chloride-induced mouse carotid artery thrombosis model indicated that fucoidan had a stronger antithrombotic activity than heparin. Further, vascular cells treated with fucoidan demonstrated a decrease in proinflammatory cytokine and chemokine production as well as inhibition of proliferation. CONCLUSION: The major findings of this study showed that fucoidan has a stronger antithrombotic effect than heparin in vivo and that fucoidan has an inhibitory effect on proinflammatory cytokine production and proliferation of vascular cells.

Study on the antithrombotic effect of fibrinolytic proteins from Scolopendra subspinipes mutilans / 中国药理学通报

Aim The aim is to purify fibrinolytic enzyme from Scolopendra subspinipes mutilans L.Koch and to study the thrombolytic and anticoagulant effect.Methods Scolopendra subspinipes mutilans L.Koch fibrinolytic enzyme(SSFE) was purified by ammonium sulphate precipitation,DEAE-cellulose and SephadexG-75 column chromatography from Scolopendra subspinipes mutilans L.Kochby.And fibrinolytic activity was determined by fibrin plate.The anticoagulant effect was measured on mice with haemolytic test and hemorrhagic test.The thrombolytic effect was measured with rats In vitro and in vivo,and the activated partial thromboplastin time(APTT),plasma prothrombin time(PT),thrombin time(TT) were measured.Results SSFE was single component with fibrinolytic activity and without any hemolyzation and hemorrhagic activity.All doses of SSFE(2,5,10 mg?kg-1) could obviously prolong activated partial thromboplastin time(APTT) and thrombin time(TT) ;Middle dose of SSFE(5 mg?kg-1) could prolong plasma prothrombin time(PT) while high dose of SSFE(10 mg?kg-1) didn't prolong obviously.Conclusion SSFE has obvious thrombolytic effect and anticoagulant effect.

Synthesis and Antithrombotic Effect of CI-930 / 第二军医大学学报

A novel cardiotonic CJ-930 was successfully designed and synthesized with a new synthetic route. The method and reactive condition on acylating, hydrolytic and condensating reaction were also improved.In addition to convenience of operation, a new synthetic route showed also the advantage over Sircar's method on the raising of total yield 26% to 36%.This compound significantly inhibited the in vitro aggregation of rabbit platelet induced by arachidonic acid, U46619, collagen, ADP and platelet activating factor(PAF). In rabbit or rat platelet, CI-930 depressed the synthesis of thromboxane A.