To explore the anti-platelet aggregation and anti-thrombotic mechanisms of Trichosanthis Fructus combined with aspirin based on network pharmacology and the validation of arteriovenous by pass model in rats. The databases of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),Drug Repositioning and Adverse Drug Reaction Chemical-Protein Interactome(DRAR-CPI),Universal Protein Resource(Uniprot) and the Database for Annotation,Visualization,and Integrated Discovery(DAVID) were used to predict protein targets and analyze biological pathway and signal pathway in the combination of Trichosanthis Fructus with aspirin. The effects of pretreatment with Trichosanthis Fructus pellets,aspirin pellets and their combination on thromboxane B2(TXB2),6-keto prostaglandin F1α(6-keto-PGF1α) and cyclic adenosine monophosphate(c AMP) in rat thrombotic model were studied. Through the study of network pharmacology,12 components of aspirin and Trichosanthis Fructus,including hydroxygenkwanin,quercetin and adenosine,were found to show the anti-platelet aggregation and anti-thrombosis mechanisms through9 common protein targets,such as SRC,RAC1,MAPK14,MAPK1,AKT1,and 14 common signaling pathways,such as VEGF signaling pathway. After the intervention with Trichosanthis Fructus pellets combined with aspirin pellets,the vascular endothslia growth factor(VEGF) signaling pathway can be activated to inhibit platelet aggregation and improve vascular endothelial function,and show the anti-platelet aggregation and anti-thrombosis mechanisms,which verify the results of the network pharmacology,and explain the anti-platelet aggregation and anti-thrombotic mechanisms of the combination of Trichosanthis Fructus pellets with aspirin pellets.
6-Ketoprostaglandin F1 alpha ; metabolism ; Animals ; Aspirin ; pharmacology ; Cyclic AMP ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Fruit ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rats ; Signal Transduction ; Thrombosis ; drug therapy ; Thromboxane B2 ; metabolism ; Trichosanthes ; chemistry

This study was designed to investigate inhibitory effects and possible mechanisms of snake venom tripeptide (pENW) on platelet adhesion in order to promote the development of a novel anti-platelet therapy. To study the inhibitory effects of pENW on platelet adhesion, washed platelets pre-incubated with pENW (116.5-466.2 μmol x L(-1)) were used to test the ability of platelet adhesion to fibrinogen. Effect of pENW on fibrin clot retraction was also tested. Effect of pENW on platelets viability was tested by MTT assay. Effect of pENW on reactive-oxygen species (ROS) levels of platelet was studied by flow cytometry assay. Calcium mobilization in Fura-2/AM-loaded platelets was monitored with a spectrofluorimeter. Cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP), thromboxane A2 (determined as its metabolite thromboxane B2) were measured using enzyme immunoassay kits. Akt, ERK and p38 phosphorylation were tested by Western blot. The results showed that pENW inhibited platelet adhesion and fibrin clot retraction in a concentration-dependent manner without cytotoxicity. Intracellular cGMP and cAMP in both resting and thrombin-activated platelets were increased by pENW. In addition, pENW attenuated intracellular Ca2+ mobilization and TXA2 production in platelets stimulated by thrombin. As shown by Western blot assay, Akt, ERK and p38 phosphorylation in thrombin-induced platelet were attenuated by pENW. However, inhibitory effects of pENW had nothing to do with ROS. Thus, pENW exhibited a significant inhibition on platelet adhesion to fibrinogen, which means pENW could block the first step of thrombosis as while as retard the more stable clot formation. The mechanisms of pENW on inhibition platelet adhesion might be related to instant regulations, such as protein kinases.
Blood Platelets ; drug effects ; Blotting, Western ; Calcium ; metabolism ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Flow Cytometry ; Phosphorylation ; Platelet Aggregation ; drug effects ; Reactive Oxygen Species ; metabolism ; Snake Venoms ; chemistry ; Thromboxane A2 ; metabolism ; Thromboxane B2 ; metabolism

This study is to investigate the effects of paeoniflorin on cerebral blood flow and the balance of PGI2/TXA2 of rats with focal cerebral ischemia-reperfusion injury. A total of 72 SD rats (3) were randomly divided into 6 groups: sham operation group, cerebral ischemia-reperfusion model group (I/R gourp), low (10 mg.kg-1), middle (20 mg.kg-1) and high (40 mg.kg-1) doses of paeoniflorin groups and nimrnodipine group. Focal cerebral ischemia in rats was made by inserting a monofilament suture into internal carotid artery for 90 min and then reperfused for 24 h. The effects of paeoniflorin on neurological deficit scores and the infarction volume of brain were detected. Relative regional cerebral blood flow (rCBF) was continuously monitored over ischemic hemispheres by laser-Doppler flowmetry (LDF). The expression of COX-2 in hippocampal CAl region was estimated by immunohistochemistry and the contents of prostacyclin I2 (PGI2), thromboxane A2 (TXA2), and ratio of PGIJ2/TXA2 in serum were measured by ELISA kits. Paeoniflorin significantly ameliorated neurological scores, reduced the infarction volume, and increased regional cerebral blood flow relative to the I/R group. In addition, paeoniflorin could inhibit COX-2 expression and the release of TXA2 and prevent the downregulation of PGI2 induced by I/R injury. The neuroprotective effects of paeoniflorin against focal cerebral ischemia-reperfusion rats might be attributed to improve the supply of injured hemisphere blood flow and adjust the balance between PGI2/TXA2.
6-Ketoprostaglandin F1 alpha ; blood ; Animals ; Brain ; blood supply ; CA1 Region, Hippocampal ; metabolism ; Cyclooxygenase 2 ; metabolism ; Glucosides ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; blood ; metabolism ; pathology ; physiopathology ; Male ; Monoterpenes ; isolation & purification ; pharmacology ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Regional Blood Flow ; drug effects ; Reperfusion Injury ; metabolism ; physiopathology ; Thromboxane B2 ; blood

OBJECTIVETo investigate the effect of atorvastatin on platelet aggregation and activation in the acute phase following balloon-induced carotid artery injury in rabbits fed cholesterol-enriched diet.

METHODSThirty rabbits were randomly divided into 5 equal groups, namely control group, high-cholesterol group, model group, low-dose (5 mg/kg daily) atorvastatin group, and high-dose (10 mg/kg daily) atorvastatin group. Platelet aggregation rate was measured in the rabbits by turbidimetric platelet aggregometry, and the changes of serum P-selectin and thromboxane B2 (TXB2) levels were detected with enzyme-linked immunosorbent assay (ELISA).

RESULTSCompared with those in the control group, serum P-selectin level increased significantly (P<0.01) but platelet aggregation rate and TXB2 level exhibited no obvious changes in high-cholesterol group. After carotid artery balloon injury, P-selectin and TXB2 levels and platelet aggregation significantly increased in cholesterol-fed rabbits, reaching the peak level at 24 h after the injury (P<0.01). Compared with the model group, low-dose atorvastatin treatment significantly decreased P-selectin and TXB2 levels and inhibited platelet aggregation in cholesterol-fed rabbits following carotid artery balloon injury (P<0.01), and such effects of atorvastatin were more prominent at a higher daily dose of 10 mg/kg (P<0.05).

CONCLUSIONSCarotid artery balloon injury in rabbits fed cholesterol-enriched diet can induce platelet activation and aggregation, which reaches the peak level at 24 h after balloon injury and can be dose-dependently inhibited by atorvastatin in the acute phase following the injury.

Animals ; Atorvastatin Calcium ; Blood Platelets ; Carotid Artery Injuries ; drug therapy ; Cholesterol ; Enzyme-Linked Immunosorbent Assay ; Heptanoic Acids ; pharmacology ; P-Selectin ; metabolism ; Platelet Activation ; Platelet Aggregation ; Pyrroles ; pharmacology ; Rabbits ; Thromboxane B2 ; metabolism

OBJECTIVETo investigate the effects of Qushuanling Capsule ( QSLC) on thrombus formation and platelet aggregation in rats.

METHODSArteriovenous bypass, venous thrombosis, and middle cerebral artery thrombosis models were used in rats to investigate the anti-thrombotic effects of QSLC, a compound of nine Chinese herbs. The platelet aggregation induced by adenosine diphosphate (ADP), thrombin or arachidonic acid (AA), as well as the contents of thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F1α (6-keto-PGF1α) in rat plasma and aortic walls, were determined to investigate the possible mechanisms of the anti-thrombotic effects of QSLC.

RESULTSAfter oral administration with QSLC for 7 days, arteriovenous bypass thrombosis was obviously suppressed compared with the model group, venous thrombosis was also obviously suppressed, rat behaviors were obviously improved, and brain infarct size as well as water content were also reduced. The platelet aggregation induced by ADP or thrombin was inhibited by QSLC, but the drug had no effect on AA-induced platelet aggregation and content of TXB(2) and 6-keto-PGF1α in plasma and the aortic wall.

CONCLUSIONThese results suggest that QSLC can be used in the prevention and treatment of thrombotic diseases, and that its mechanism of action may be related to inhibition of platelet aggregation.

6-Ketoprostaglandin F1 alpha ; blood ; Adenosine Diphosphate ; pharmacology ; Animals ; Aorta ; drug effects ; metabolism ; pathology ; Cerebral Infarction ; blood ; drug therapy ; pathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Middle Cerebral Artery ; drug effects ; pathology ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Thrombosis ; drug therapy ; pathology ; Thromboxane B2 ; blood ; Venous Thrombosis ; drug therapy ; pathology

OBJECTIVETo evaluate the effect of peroxisome proliferator-activated receptor (PPAR)α agonist fenofibrate on the secretion of endothelium-derived contracting factors in hypertensive rats.

METHODSThe changes of vascular tension in SHR rats after having been incubated with 0.1, 1.0, or 10.0 μmol/L fenofibrate or 10.0 μmol/L fenofibrate and PPARα antagonist MK866 or PPARγ antagonist GW9662 for one hour were observed, and the findings were compared with those in WKY rats (control group). The serum levels of vascular endothelial contraction factor prostacyclin (PGF) 1α, 2α, and thromboxane B2 (TXB2) were determined by enzyme-linked immunosorbent assay (ELISA). The expression of COX-1 protein was determined by Western blot analysis.

RESULTSCompared with the control group, fenofibrate significantly reduced the vasoconstriction ability of the SHR rats(P=0.013). PPARα antagonist MK866 significantly improved the vascular contractility of SHR rats that had been incubated with 10.0 μmol/L fenofibrate (P=0.021). PPARγ antagonist GW9662 had no significant effect on the vascular contractility of SHR rats after having been incubated with 10.0 μmol/L fenofibrate (P=0.071). The serum levels of PGF1α(P=0.014), 2α(P=0.023), and TXB2 (P=0.017) in SHR rats incubated with 10.0 μmol/L fenofibrate were significantly lower than in the control group. With the presence of vascular endothelium, the expression of COX-1 in SHR rats incubated with fenofibrate was significantly lower than that in SHR rats incubated without fenofibrate (P=0.027).

CONCLUSIONFenofibrate reduces the secretion of endothelium-dependent contracting factors in SHR rats through lowering the expression of COX-1.

Animals ; Cyclooxygenase 1 ; metabolism ; Epoprostenol ; metabolism ; Fenofibrate ; pharmacology ; Hypertension ; metabolism ; Male ; Membrane Proteins ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Thromboxane B2 ; metabolism

Effects of the effective components group of Xiaoshuantongluo formula (XECG) on rat acute blood stasis model were studied under the guidance of the concept of effective components group. Rat acute blood stasis model was induced by subcutaneous injection of epinephrine combined with ice water bath. Hemorheology indices such as whole blood viscosity, plasma viscosity, erythrocyte aggregation index and platelet aggregation rate; coagulation parameters including PT, APTT, TT and FIB; 6-keto-PGF1alpha, TXB2 and D-dimer levels were determined to evaluate the effects of XECG. The results showed that XECG significantly reduced ADP-induced platelet aggregation, but showed little influence on the whole blood viscosity, plasma viscosity and erythrocyte aggregation rate. XECG extended PT and TT slightly, but had no effects on APTT and FIB content. D-dimer levels significantly decreased after administration of XECG with a little decrease of TXB2, but the content of 6-keto-PGF1alpha did not change significantly. The results suggest that the role of XECG of anti-aggregation is more prominent.
6-Ketoprostaglandin F1 alpha ; blood ; Animals ; Blood Coagulation ; drug effects ; Blood Coagulation Disorders ; blood ; Blood Viscosity ; drug effects ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocyte Aggregation ; drug effects ; Fibrin Fibrinogen Degradation Products ; metabolism ; Hemorheology ; drug effects ; Male ; Partial Thromboplastin Time ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Prothrombin Time ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thrombin Time ; Thromboxane B2 ; blood

Protective effects of two different extracts of TSCA (total saponins from Cicer arietinum) were studied on kidney of T2DM rats. The diabetic model group was established with high calorie feeding and small dose injection of streptozotocin (STZ, 45 mg x kg(-1)). DM rats were randomly assigned to model group (feed with propylene glycol 1 mL/100 g), TSCA high dose group (300 mg x kg(-1)), TSCA low dose group (100 mg x kg(-1)) and normal control group (feed with propylene glycol 1 mL/100 g). After four weeks treatment with TSCA I and II, the levels of FPG FIns, BUN, Scr, ATII, ET-1, TXB2 and 6-keto-PGF1alpha in blood and the activities of SOD, GSH-PX and MDA in kidney were analyzed by biochemical methods. After four weeks treatment with TSCA II, the levels of FPG FIns, BUN, Scr, ATII and ET-1 were reduced significantly; and the ratios of TXB2 to 6-keto-PGF1alpha and SOD were effectively alleviated in TSCA II group. While there is no significant change on FPG and BUN in comparison to the rats treated with TSCA I, Scr, ATII, ET-I, GSH-PX and SOD were alleviated. The results suggest that TSCA II could be used to reduce FPG and FIns. According to the result of vasoactive substances index, TSCA II is more effective than TSCA I on renal protection of DM rats.
6-Ketoprostaglandin F1 alpha ; blood ; Angiotensin II ; blood ; Animals ; Blood Glucose ; metabolism ; Blood Urea Nitrogen ; Cicer ; chemistry ; Creatinine ; blood ; Diabetes Mellitus, Experimental ; blood ; metabolism ; Diabetes Mellitus, Type 2 ; blood ; metabolism ; Endothelin-1 ; blood ; Glutathione Peroxidase ; metabolism ; Hypoglycemic Agents ; isolation & purification ; pharmacology ; Insulin ; blood ; Kidney ; metabolism ; Male ; Malondialdehyde ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology ; Superoxide Dismutase ; metabolism ; Thromboxane B2 ; blood

OBJECTIVETo investigate the changes in contents of cycloxygenase-2 (COX-2) and its downstream effectors in rat's myocardial ischemia/reperfusion (I/R) model and observe the effects of precondition with GBE50 (Ginkgo biloba extract 50) and Salviae miltiorrhizae (SM) on them.

METHODSRat's I/R model was established by 30-min left anterior descending coronary artery occlusion followed with 60-min reperfusion. Animals were divided into the model control group, the sham-operated group and the tested groups (received 1-week precondition with GBE50 and SM respectively via intragastric infusion before modeling). COX-2 mRNA expression in myocardium was detected by real-time PCR; contents of thromboxane B2 (TXB2) and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) were measured by radioimmunoassay.

RESULTSThe mRNA expression of COX-2 in the model group was obviously higher than that in the sham-operated group (P < 0.001), while that in the tested groups was down-regulated significantly (P < 0.01), and the content of TXB2 as well as the ratio of TXB2/PGF1alpha was reduced significantly (P < 0.05). Besides, SM also showed the up-regulation effect on 6-keto-PGF1alpha content in myocardium (P < 0.05).

CONCLUSIONCOX-2 affects the myocardium through thromboxane A2 and prostacyclin after I/R; both GBE50 and SM can inhibit the production of COX-2, but they may act in different paths.

6-Ketoprostaglandin F1 alpha ; metabolism ; Animals ; Cyclooxygenase 2 ; genetics ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Ginkgo biloba ; chemistry ; Ischemic Preconditioning, Myocardial ; methods ; Male ; Myocardial Ischemia ; physiopathology ; Myocardial Reperfusion Injury ; metabolism ; pathology ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Thromboxane B2 ; metabolism

OBJECTIVETo observe the effect of a integrative medical regimen (IMR), i.e. combined use of Jiangya Capsule (JYC) and Nimodipine (ND), on blood pressure, TCM clinical symptoms, and blood levels of vascular endothelial function and hypersensitive C-reactive protein (hs-CRP) in elderly patients with isolated systolic hypertension (EISH).

METHODSAdopting randomized, double-blinded and controlled principle, a trial was conducted on 135 patients with EISH by randomized them into three groups, they were administered IMR (Group A), JYC plus ND simulator (Group B) and ND plus JYC simulator (Group C) respectively, for 4 weeks. Changes of blood pressure and TCM symptoms, as well as the levels of serum nitric oxide (NO), plasma endothelin-1 (ET-1), 6-keto-prostaglandin 1alpha (6-keto-PGF(1alpha)), thromboxane B2 (TXB2) and hs-CRP were observed before and after treatment.

RESULTSAfter treatment the systolic blood pressure reduced and clinical symptoms improved, with serum NO and 6-keto-PGF(1alpha) lelels elevated, plasma ET-1, TXB2 and serum hs-CRP decreased in all the three groups (P<0.05 or P<0.01). But the inter-group comparisons showed that the effect in Group A was superior to the other two groups in decreasing systolic pressure, and superior to Group C in improving clinical symptoms, elevating serum NO and decreasing plasma TXB2 (P <0.05).

CONCLUSIONThe integrative medical regimen of combined use JYC and ND has markedly effect in lowering blood pressure, it could obviously improve the symptoms and vascular endothelial function, and inhibit the level of inflammatory factor in patients with EISH.

Aged ; C-Reactive Protein ; metabolism ; Double-Blind Method ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Endothelin-1 ; blood ; Female ; Humans ; Hypertension ; drug therapy ; Integrative Medicine ; Male ; Middle Aged ; Nimodipine ; therapeutic use ; Nitric Oxide ; blood ; Phytotherapy ; Thromboxane B2 ; blood