Clopidogrel-herb Interactions: A Pharmacokinetic and Pharmacodynamic Assessment in a Rat Model.

BACKGROUND: Herbs usually contain a mixture of biologically active constituents, which can interact with numerous prescribed drugs and alter their safety profiles. OBJECTIVES: The current investigation was aimed to evaluate the effect of commonly used herbal products including black seed (Nigella sativa), garden cress (Lepidium sativum), and fenugreek (Trigonella foenum-graecum) on the pharmacokinetics and pharmacodynamics of clopidogrel using a Wistar rat model. METHODS: A GC-MS analysis revealed the presence of several phytoconstitutents (polyphenols) in the extracts of black seed, garden cress, and fenugreek. These polyphenols have the potential to interfere with clopidogrel effect. Plasma concentrations of clopidogrel were measured at different time points in the absence and presence of the concurrent use of tested herbal products and the pharmacokinetic parameters were calculated. Bleeding time was measured in various groups as a measure of the antiplatelet effect of clopidogrel. RESULTS: Area under the plasma concentration-time curves (AUC0-∞) of clopidogrel were 35.53 ±0.89 µg/ml*h (p<0.05), 26.01 ±0.90 µg/ml*h (p>0.05) and 32.80 ±2.51 µg/ml*h (p<0.05) in the black seed, garden cress and fenugreek group, respectively, compared with that of the control group (27.02 ±0.42 µg/ml*h). Treatment with black seed also caused an increase in clopidogrel Cmax by 31.52% (p<0.05) and with fenugreek by 21.42% (p<0.05); Cmax, did not changed with garden cress treatment (6.48 ±0.15 µg/ml versus 6.12 ±0.21 µg/ml, p>0.05). The pharmacodynamic evaluation of the antiplatelet effect of clopidogrel in the presence of herbal products treatment showed a significant prolongation in the bleeding time from a control baseline by ~22-26%, and by added ~8-12% in reference to clopidogrel therapeutic effect (p<0.05). CONCLUSION: The concurrent use of black seed, fenugreek, or garden cress can alter the pharmacokinetics and pharmacodynamics of clopidogrel to varying degrees due to the presence of various bioactive polyphenols. This is probably due to changes in drug disposition and its antiplatelet action. Further confirmation can determine the clinical relevance of these observations and identify the exact constituents responsible for such activities.

Gene therapy for hemophilia B using CB 2679d-GT: a novel factor IX variant with higher potency than factor IX Padua.

Sustained expression of therapeutic factor IX (FIX) levels has been achieved after adeno-associated viral (AAV) vector-based gene therapy in patients with hemophilia B. Nevertheless, patients are still at risk of vector dose-limiting toxicity, particularly liver inflammation, justifying the need for more efficient vectors and a lower dosing regimen. A novel increased potency FIX (designated as CB 2679d-GT), containing 3 amino acid substitutions (R318Y, R338E, T343R), significantly outperformed the R338L-Padua variant after gene therapy. CB 2679d-GT demonstrated a statistically significant approximately threefold improvement in clotting activity when compared with R338L-Padua after AAV-based gene therapy in hemophilic mice. Moreover, CB 2679d-GT gene therapy showed significantly reduced bleeding time (approximately fivefold to eightfold) and total blood loss volume (approximately fourfold) compared with mice treated with the R338L-Padua, thus achieving more rapid and robust hemostatic correction. FIX expression was sustained for at least 20 weeks with both CB 2679d-GT and R338L-Padua whereas immunogenicity was not significantly increased. This is a novel gene therapy study demonstrating the superiority of CB 2679d-GT, highlighting its potential to obtain higher FIX activity levels and superior hemostatic efficacy following AAV-directed gene therapy in hemophilia B patients than what is currently achievable with the R338L-Padua variant.

Effects of Algan Hemostatic Agent on bleeding time in a rat tail hemorrhage model.

BACKGROUND: Algan Hemostatic Agent (AHA) is a multi-herbal extract containing a standardized amount of Achillea millefolium, Juglans regia, Lycopodium clavatum, Rubus caesius or Rubis fruciosus, Viscum album, and Vitis vinifera, each of which is effective in hemostasis. In this study, we aimed to investigate the effects of AHA on bleeding time in a rat tail hemorrhage model. METHODS: Forty-eight Sprague Dawley rats (5-7 weeks old, 180-210 g) were randomly and equally allocated to six groups as follows: heparin plus saline (heparinized control), heparin plus AHA-soaked sponge, heparin plus liquid form of AHA, saline (non-heparinized control), AHA-soaked sponge and liquid form of AHA. Heparin (640 IU/kg) was administered intraperitoneally three times a day for three days in heparinized groups. For the bleeding model, the tail of rats was transected. According to the study group, either saline- or AHA-soaked sponge or liquid form of AHA was applied over the hemorrhage area. In AHA- or saline-soaked sponge groups, once the bleeding time had started, it was checked every 10 seconds. If the bleeding did not stop after 40 seconds, it was accepted as a failure. In liquid AHA group, the duration of bleeding was measured using a chronometer and defined as the time (seconds) from wounding until the bleeding stopped. RESULTS: Bleeding time in the heparinized and non-heparinized control groups was over 40 seconds. After applying the sponge form of AHA on the wound area, bleeding time was significantly shortened to less than 20 seconds in both heparinized and non-heparinized rats (p<0.001 for both). The liquid form of AHA stopped bleeding in 5.0±1.2 seconds and 8.0±1.3 seconds in heparinized and non-heparinized groups, respectively. CONCLUSION: AHA is a highly effective topical hemostatic agent in a rat tail hemorrhage model, thus may provide for a unique clinically effective option for control of bleeding during surgical operations or other emergencies.

Hemostatic activity of aqueous extract of Myrtus communis L. leaf in topical formulation: In vivo and in vitro evaluations.

ETHNOPHARMACOLOGICAL RELEVANCE: Myrtus communis L. (MC) is a well-known medicinal plant in traditional Persian medicine, which contains a large amount of phenolic compounds (mainly hydrolyzable tannins). As mentioned in ancient literature, MC was widely used to control bleeding in every part of the body. Nevertheless, there is no pharmacological study on the anti-hemorrhagic activity of this plant till now. AIM OF THE STUDY: The current in vivo and in vitro study aimed at evaluating the hemostatic activity of M. communis aqueous leaf extract (MCE) in topical formulation. MATERIALS AND METHODS: Two parameters of bleeding time and amount in tail bleeding model were measured in vivo in rats treated with MCE (1%, 2.5%, 5%, 7.5%, 10%, 15%, and 20% w/v), 5% M. communis aqueous leaf extract gel (G), tannic acid (TA) (1%, 2.5%, 5%, 7.5%, and 10%), normal saline (NS), and the Monsel's solution (MS), a commercial hemostatic agent. Also, the effect of 5% MCE and 5% TA on PT (prothrombin time) and aPTT (activated partial thromboplastin time) as well as protein precipitation and platelet aggregation were assessed in vitro. RESULTS: In the rat-tail bleeding model, bleeding time and amount significantly (P < 0.001) reduced by the application of 5% MCE solution on the cut tail compared with the NS group. The bleeding time and amount in the MS group were not significantly different from those of the 5% MCE group. Platelet microaggregates were detected by fluorescent microscope. PT and aPTT values increased >120 s and >180 s by 5% MCE, respectively. Also, protein precipitation and significant reduction in serum proteins were observed in the 5% MCE group. CONCLUSION: The current study provided new insights into the hemostatic effect of MCE, which may be partially mediated by platelet aggregation activity. Hence, it could be evaluated as the resource of new plant origin hemostatic agent.

Interactions between clopidogrel and traditional Chinese medicine.

The use of traditional Chinese medicine (TCM) has obtained more and more acceptance all over the world due to its multi-target and multi-level function characteristics. Clopidogrel is a major therapeutic option to reduce atherothrombotic events in patients with acute coronary syndrome, recent myocardial infarction, recent stroke or established peripheral arterial disease. These patients probably take TCM. Are there any interactions between clopidogrel and TCM? Whether TCM will affect the efficacy of clopidogrel or increase the adverse reactions of bleeding? Clarifying this information will help physicians make better use of TCM. A literature search was carried out using Web of Science, PubMed and the Cochrane Library to analyze the pharmacokinetic or pharmacodynamic interactions of clopidogrel and TCM. Some herbs can increase the AUC or Cmax of clopidogrel, such as Scutellarin, Danggui, Gegen, Sauchinone and Dengzhan Shengmai capsules. Whereas others can decrease clopidogrel, for example, Ginkgo and Danshen. Furthermore, some herbs can increase the AUC or Cmax of clopidogrel active metabolite, including Ginkgo and Xuesaitong tablet. And others can decrease the clopidogrel active metabolite, such as Scutellarin, Danshen, Fufang Danshen Dripping Pill and Dengzhan Shengmai capsules. Additionally, Schisandra chinensis, Danggui, Gegen and Fufang Danshen Dripping Pill can decrease the AUC or Cmax of the clopidogrel inactive metabolite, while Curcumin on the contrary. The pharmacodynamics of Panax notoginseng, Notoginsenoside Ft1, Hypericum perforatum, Shexiang baoxin pills, Naoxintong capsule increased the antiplatelet activity compared with clopidogrel alone, while Danshen decreased the platelet inhibition. In adverse reactions, Danggui can enhance the adverse effects of clopidogrel on the bleeding time. With more awareness and understanding on potential drug-herb interactions of clopidogrel and TCM, it may be possible to combine clopidogrel with TCM herbs to yield a better therapeutic outcome.

In Vitro Effects on Thrombin of Paris Saponins and In Vivo Hemostatic Activity Evaluation of Paris fargesii var. brevipetala.

The resource shortage of Rhizoma Paridis has never been effectively addressed, and the industry continues to search for alternative resources. The in vitro effects on thrombin of Paris saponins and in vivo hemostatic activity of Paris fargesii var. brevipetala (PF) were evaluated in this study. PF is considered to be an alternative source of Rhizoma Paridis (RP). The in vitro incubation experiment was designed to investigate the effects on thrombin activity of Paris saponin H (PS H) and saponin extract in PF. The bleeding time of mouse tail snipping was used to evaluate the in vivo hemostatic effects of Paris saponins. Also, in vivo changes in four blood coagulation parameters in rats after oral administration of different groups of Paris saponins were compared. The effects of Paris saponins on liver function and blood lipid parameters were examined in order to avoid drug-induced liver injury. Activity studies of thrombin after ultra-filtration centrifugation showed that Paris saponins were able to enhance thrombin activity. Ultra performance liquid chromatography mass spectrometry (UPLC-MS) analysis results of the substrates led us to speculate that there is a specific binding between Paris saponins and thrombin. PS H and Paris saponins in PF significantly shortened the bleeding time in mice. One pathway by which Paris saponins enhance in vivo blood coagulation is by increasing fibrinogen (FIB), among the four blood coagulation parameters in rats. At the same time, the effects on liver and blood lipid parameters were insignificant. P. fargesii var. brevipetala can be developed as an alternative medicinal source of Rhizoma Paridis.

Comparing coagulation activity of Selaginella tamariscina before and after stir-frying process and determining the possible active constituents based on compositional variation.

CONTEXT: Selaginella tamariscina (P. Beauv.) Spring (Selaginellaceae) (ST) has been widely used in China as a medicine for improving blood circulation. However, its processed product, S. tamariscina carbonisatus (STC), possesses opposite haemostatic activity. OBJECTIVE: To comprehensively evaluate the activity of ST and STC on physiological coagulation system of rats, and seek potential active substances accounting for the activity transformation of ST during processing. MATERIALS AND METHODS: The 75% methanol extracts of the whole grass (fine powder) of ST and STC were prepared, respectively. Male Sprague-Dawley rats were randomly divided into five groups: control group, model group, model + ST group, model + STC group and positive control group (model + Yunnanbaiyao). The duration of intragastric administration was 72 h at 12 h intervals. Haemorheology parameters were measured using an LB-2 A cone-plate viscometer and the existed classic methods, respectively. SC40 semi-automatic coagulation analyzer was employed to determine coagulation indices. Meanwhile, HPLC and LC-MS were applied for chemical analyses of ST and STC extracts. RESULTS: STC shortened tail-bleeding time, increased whole blood viscosity (WBV) and plasma viscosity (PV), decreased erythrocyte sedimentation rate blood (ESR), reduced activated partial thromboplastin time (APTT) and increased the fibrinogen (FIB) content in the plasma of bleeding model rats. Although ST could shorten APTT and TT, the FIB content was significantly decreased by ST. Dihydrocaffeic acid with increased content in STC vs. ST showed haemostatic activity for promoting the platelet aggregation induced by collagen and trap-6, and reducing APTT and PT significantly with a concentration of 171.7 µM in vitro. Amentoflavone with reduced content in STC vs. ST inhibited ADP and AA-induced platelet aggregation significantly with a concentration of 40.7 µM. DISCUSSION AND CONCLUSIONS: As the processed product of ST, STC showed strong haemostatic activity on bleeding rat through regulating the parameters involved in haemorheology and plasma coagulation system. Two active compounds, dihydrocaffeic acid and amentoflavone, might be partially responsible for the haemostatic and anticoagulant activity of STC and ST, respectively.

Novel carbon dots derived from Schizonepetae Herba Carbonisata and investigation of their haemostatic efficacy.

Schizonepetae Herba Carbonisata (SHC) has been used in traditional Chinese medicine (TCM) to treat haemorrhagic diseases for more than 1000 years. However, little information is available on its haemostatic components and mechanism. In this study, we developed novel water-soluble carbon dots (CDs) in aqueous extracts of SHC for the first time and a modified pyrolysis method was used to prepare the SHC using Schizonepetae Herba (SH) as the sole precursor. The SHC-CDs were characterized using transmission electron microscopy (TEM), high-resolution TEM (HRTEM), Fourier transform infrared (FT-IR), ultraviolet-visible (UV-Vis) and fluorescence spectroscopy, X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and high-performance liquid chromatography (HPLC). Furthermore, the CDs with a quantum yield (QY) around 2.26% exhibited no toxicity within approximately 0.84 mg/mL in the CCK-8 assay. More interestingly, tail haemorrhaging and liver haemorrhaging experiments showed that CDs-treated mice had significantly shorter bleeding time than did normal saline (NS)-treated control group. Coagulation assays suggested that SHC-CDs could stimulate the extrinsic blood coagulation system and activate the fibrinogen system. These results suggested that SHC-CDs possess a remarkable haemostatic property, which provides evidence to support the further investigation of the considerable potential and effective material basis of TCM.

Antiplatelets and profibrinolytic activity of Citrullus colocynthis in control and high-fat diet-induced obese rats: mechanisms of action.

The current study aimed to investigate the effect of Citrullus colocynthis (C. colocynthis) hydro-alcoholic extract on blood haemostasis in control and high-fat diet (HFD) induced obese rats. In control rats, the extract significantly enhanced bleeding time and plasma levels of tPA and significantly decreased plasma levels PAI-1 and serum levels of thromboxane B2 leading to inhibition of platelets aggregation. In HFD induced obese rats, similar effects were seen and the extract was also able to reverse HFD induced increases in fibrinogen and VWF. Searching for the mechanism, C. colocynthis acts by (1) inhibiting of food intake, (2) inhibiting the activity of pancreatic lipase, (3) decreasing levels of TNF-α and IL-6 and (4) decreasing circulatory levels of the prothrombotic adipokine, leptin and enhanced circulatory levels of the antithrombic adipokines and adiopnectin. In conclusion, C. colocynthis has antiplatelets and profibrinolytic activity in both control and HFD induced obese rats.

Antithrombotic Effect of Fermented Ophiopogon japonicus in Thrombosis-Induced Rat Models.

In this study, the antithrombotic and thrombolytic ability of second fermented extract of Ophiopogon japonicus (FEOJ) was verified in thrombosis-induced rats. Thrombosis was induced by oral administration of 2% carrageenan for 4 weeks. Five experimental groups (n = 9/group) involved in the study were control group, thrombosis group, low-dose FEOJ group (2 mL/kg, low-dose Ophiopogon japonicus [LOJ]), middle-dose FEOJ group (6 mL/kg, medium-dose Ophiopogon japonicus [MOJ]), and high-dose FEOJ group (12 mL/kg, high-dose Ophiopogon japonicus [HOJ]). The clotting time (CT), bleeding time (BT), prothrombin time (PT), activated partial thromboplastin time (APTT), and fibrinogen (FBG) were assessed in blood samples, and histological studies were performed on liver and lung tissues. The results demonstrated delayed CT only in MOJ and HOJ groups and delayed BT in all FEOJ groups compared with those in thrombosis and control groups (P < .05). Similarly, APTT was significantly delayed only in MOJ and HOJ groups, and PT was significantly delayed in all FEOJ groups, compared with those in control and thrombosis groups (P < .05). Although concentrations of FBG were similar in control, thrombosis, and LOJ groups, the tendency for decreased concentration of FBG (statistically nonsignificant) in MOJ and HOJ groups has been observed. Histological examination of livers and lungs revealed that thrombosis was partially improved in FEOJ group compared with the thrombosis group. In conclusion, CT, BT, PT, and APTT were prolonged in FEOJ group more than in control and thrombosis groups, thereby, depicting antithrombotic and thrombolytic effects. However, concentration-dependent effects of FEOJ were more prominent in MOJ and HOJ groups than in the LOJ group.

The effects of oral administration of Yunnan Baiyao on blood coagulation in beagle dogs as measured by kaolin-activated thromboelastography and buccal mucosal bleeding times.

We examined the effects of oral administration of Yunnan Baiyao (YB) on hemostasis by measuring buccal mucosal bleeding times (BMBTs) and doing citrated kaolin-activated whole-blood thromboelastography (TEG). In a randomized controlled crossover trial 8 beagle dogs were given either placebo or 1000 mg of YB orally every 12 h for 5 consecutive treatments. Blood was drawn 24 h before treatment and 2 and 24 h after the last treatment, and the BMBT was measured in each sample in duplicate. The TEG analysis was done in duplicate 60 ± 5 min after sample collection. There were no adverse effects of treatment and no significant differences between the control and treatment BMBTs or TEG parameters at any time point. Significant differences were found between baseline and 24 h after the last treatment within the treatment group for the TEG parameters LY30 and LY60 and within the control group for the TEG parameters MA, G, LY30, and LY60. Thus, at the dose and frequency of administration in this study YB did not appear to have any clinically significant effects on the measured coagulation parameters. The differences within the treatment group were likely due to analytic error since similar differences were seen in the control group. Further studies with a larger sample, as well as more direct measures of platelet function, are needed.

Nous avons examiné les effets de l'administration orale de Yunnan Baiyao (YB) sur l'hémostase en mesurant le temps de saignement de la muqueuse buccale (TSMB) et en faisant une thromboélastographie (TEG) de sang entier après activation par de la kaoline citratée. Lors d'un essai en croisé randomisé et contrôlé, huit chiens beagle ont reçu soit un placebo ou 1000 mg de YB par voie orale à chaque 12 h pour cinq traitements consécutifs. Du sang a été prélevé 24 h avant le traitement et 2 et 24 h après le dernier traitement, et le TSMB mesuré dans chaque échantillon en duplicata. L'analyse TEG a été faite en duplicata 60 ± 5 min après le prélèvement de l'échantillon. Il n'y eut aucun effet néfaste du traitement et aucune différence significative entre le groupe témoin et le groupe traité pour ce qui est des TSMBs ou des paramètres de la TEG à tous les points d'échantillonnage. Des différences significatives ont été trouvées entre les valeurs de base et 24 h après le dernier traitement à l'intérieur du groupe traité pour les paramètres LY30 et LY60 de la TEG et à l'intérieur du groupe témoin pour les paramètres MA, G, LY30 et LY60 de la TEG. Ainsi, à la dose et à la fréquence d'administration utilisées dans la présente étude, YB ne semble pas avoir d'effet clinique significatif sur les paramètres de coagulation mesurés. Les différences dans le groupe traité sont fort probablement dues à une erreur analytique car des différences similaires ont été notées dans le groupe témoin. Des études supplémentaires avec un échantillonnage plus grand, ainsi que des mesures plus directes de la fonction des plaquettes sont requises.(Traduit par Docteur Serge Messier).

Atractylodes lactone compounds inhibit platelet activation.

Platelets play a crucial role in the development and progression of atherosclerosis-thrombosis and, therefore, antiplatelet drugs are widely used in the treatment of coronary artery disease. Moreover, advances in understanding the biological functions of natural plant products can provide new pharmacological strategies aimed at promoting cardiovascular health. Atractylenolide I (ATL-1), ATL-2, and ATL-3 are the major bioactive components of a Qi tonifying medicinal herb Rhizoma Atractylodis Macrocephalae (Atractylodes macrocephala), which is commonly used in traditional Chinese medicine (TCM). These components possess well-documented anti-inflammatory and anticancer activities, but their effects on platelet activation are still unknown. In this study, the effects of ATL on platelet function in vitro and in vivo were investigated, and the underlying mechanism was explored. We found that ATL-2 and ATL-3 but not ATL-1 diminished agonist-induced platelet aggregation and diminished adenosine triphosphate (ATP) release from dense granules. The levels of phospho-Akt (Ser473) and phospho-p38 MAPK were downregulated in the presence of ATL-2 and ATL-3. We also found that ATL-2 and ATL-3 have a similar inhibitory effect on platelet activation as acetylsalicylic acid in response to agonists. Furthermore, ATL-2 and ATL-3 diminished the spreading of human platelets on immobilized fibrinogen (Fg), delayed clot retraction in platelet-depleted plasma containing human platelets, extended first occlusion time in a mouse model of ferric chloride (FeCl3)-induced carotid arterial thrombosis, and prolonged the bleeding time. These observations suggest that ATL-2 and ATL-3 are potential candidate therapeutic drugs for the prevention of thrombosis.

Anti-platelet and anti-thrombotic effect of a traditional herbal medicine Kyung-Ok-Ko.

ETHNOPHARMACOLOGICAL RELEVANCE: Kyung-Ok-Ko (KOK), a traditional herbal prescription, contains six main ingredients; Rehmannia glutinosa var. purpurae, Lycium chinense, Aquillaria agallocha, Poria cocos, Panax ginseng, and honey. KOK has been widely taken as a traditional oriental medicine for improving blood circulation or age-related symptoms, such as dementia and stroke. However, the effect of KOK on platelet activity has not been clarified. MATERIALS AND METHODS: To evaluate the effect of KOK on platelet function, we evaluated its effect on functional markers of platelet activation such as aggregation and shape change. As a mechanism study for the effect of KOK, we examined its effect on granule secretion, intracellular Ca(2+) increase, and PLCγ and Akt activation. To investigate the effect of orally administered KOK (0.5, 1, 2 g/kg), we examined its ex vivo effect on platelet aggregation in rat, and its in vivo anti-thrombotic effect in mice thromboembolism model. Furthermore, the effect of KOK on bleeding time was examined to estimate its potential side effect. RESULTS: KOK (0.3, 1, 3, 10 mg/ml) inhibited collagen-induced platelet aggregation and shape change in rat platelets in a concentration-dependent manner. The mechanism for the anti-platelet effect of KOK seems to involve the inhibition of ATP release, intracellular Ca(2+) elevation, and the phosphorylation of PLCγ and Akt. In rat ex vivo study, KOK (2 g/kg, p.o. for 1 day, and 0.5, 1, 2 g/kg, p.o. for 7 days) also had significant inhibitory effects on collagen-induced platelet aggregation. In addition, KOK showed a significant protective effect against thrombosis attack in mice. The prolongation of bleeding time by KOK was much less than that by ASA, suggesting a beneficial potential of KOK than ASA in view of side effect. CONCLUSIONS: These findings suggest that KOK elicits remarkable anti-platelet and anti-thrombotic effects with less side effect of bleeding, and therefore, it may have a therapeutic potential for the prevention of platelet-associated cardiovascular diseases.

Comparison of microporous polysaccharide hemospheres and Ankaferd Blood Stopper in a rabbit epistaxis model.

The aim of this study was to evaluate the histopathological impact, effectiveness, and safety of two hemostatic agents, Ankaferd Blood Stopper (ABS) and microporous polysaccharide hemospheres (MPH), in an experimental rabbit epistaxis model. Rabbits were randomly assigned, using a computerized random number generator, to the following three groups of six animals: group 1 (control, irrigated with saline); group 2 (ABS-treated); and group 3 (MPH-treated). In all groups, a standardized rabbit epistaxis model was used. Hemostasis time and extent of nasal bleeding were measured to compare the hemostatic effect of ABS and MPH among groups. Septums were removed for histopathological analysis, 7 days after the procedure. ABS reduced hemostasis time to 104.2 s and amount of bleeding to 20.5 mg. MPH reduced hemostasis time to 71.7 s and amount of bleeding to 11.5 mg. Mean bleeding time in wounds administered ABS and MPH was significantly shorter compared with wounds administered isotonic saline solution (p = 0.004). ABS and MPH application decreased bleeding significantly compared with the control group (p = 0.004). Bleeding time and amount in the MPH group was significantly reduced compared with the ABS group (p = 0.013 and p = 0.004, respectively). There was no significant difference in the histopathological evaluation results between the ABS, MPH, and control groups. Our data indicate that both ABS and MPH represent safe, effective, and fast-acting hemostatic agents in the management of epistaxis. MPH was more effective than ABS in terms of hemostasis time and amount of bleeding.

Effects of Ankaferd Blood Stopper and calcium alginate in experimental model of hepatic parenchymal bleeding.

BACKGROUND: This experimental study compared the hemostatic effects of calcium alginate and Anka-ferd Blood Stopper in hepatic parenchymal bleedings. MATERIAL AND METHOD: The study comprised 39 male Wistar albino rats (weight 230±30 g). Laceration model was created in the left lateral lobe of the liver. Standard cotton gauze that was impregnated 0.9% NaCl solution and Calcium alginate cover was compared to ABS tampon. The amount of preoperative bleeding, preoperative and postoperative Day 1 hematocrit levels, and the difference between them were assessed and statistically analyzed. RESULTS: Comparing the hematocrit levels between the groups, we found that the amount of bleeding was significantly higher in the control group versus the study groups (p<0.001). Histopathological examination revealed the portal area enlargement and biliary canaliculi proliferation. In the Ca2+ Alginate group, it was observed that the fibres were still present in the incision line with massive fibrotic area around. In the Ankaferd group, examination of the preparations revealed patchy focal necrosis areas but no fibrotic area. CONCLUSION: With this study, we demonstrated that both calcium alginate and Ankaferd have hemostatic effect in preventing hepatic parenchymal bleeding and that calcium alginate causes fibrosis in the liver, where ABS causes focal necrosis areas(Tab. 2, Fig. 4, Ref. 19).

Chloroform fraction of Euphorbia maculata has antiplatelet activity via suppressing thromboxane B2 formation.

Euphorbia maculata (EM) is a traditionally used antidiarrheal, antibacterial, antifungal and antioxidant agent. However, the effects of EM on platelet activity remain to be elucidated. Therefore, the present study investigated the antiplatelet effect of various EM extract fractions on platelet aggregation in rats. The antiplatelet activity of the EM fractions on collagen or adenosine diphosphate (ADP)­induced platelet aggregation was evaluated in vitro and ex vivo. Thromboxane B2 (TXB2) formation, rat­tail bleeding time and coagulation time were also measured. Among the fractions, the chloroform fraction of EM (CFEM) significantly inhibited ADP­induced platelet aggregation in vitro. Furthermore, oral administration of 50 mg/kg CFEM to rats significantly reduced ADP­induced platelet aggregation without increasing the tail bleeding time or coagulation time. In addition, EM significantly inhibited the level of TXB2 formation in a dose­dependent manner. These results suggest that CFEM exhibits antiplatelet activity, without causing bleeding, via the suppression of TXB2 formation. CFEM may be a type of food which has the potential for preventing cardiovascular disease.

Cardiovascular effects of Phaleria macrocarpa extracts combined with mainstay FAC regimen for breast cancer.

DLBS1425 is a bioactive compound extracted from Phaleria macrocarpa, with anti-proliferative, anti-inflammatory and anti-angiogenic properties against cancer cells. The present study was aimed to assess cardiotoxicity of DLBS1425, compared to the mainstay regimen for breast cancer, 5-fluorouracil:doxorubicin:cyclophosphamide (FAC, given at 500/50/500 mg/m(2)). Treatment with FAC regimen at standard dose resulted in very severe toxicity, so mice had no chance to survive for more than 7 days following initial drug treatment. Furthermore, histological examination on the heart revealed severe muscular damage when mice were given the FAC regimen alone (severe toxicity). FAC as chemotherapeutic regimen exerted high toxicity profile to the cardiovascular cells in this experiment. Meanwhile, treatment with DLBS1425 alone up to a dose equivalent to as high as 300 mg three times daily in human had no hazardous consequences on the heart, hematological feature, as well as general safety. In the cardiovascular cells, DLBS1425 in the presence of FAC regimen (one-eight of the initial dose) gave protection to the cardiac muscle cells as well as other hematological features. Taken together, results of the present study suggest that DLBS1425 is safe when used as adjuvant therapy for breast cancer and may be even protective against cardiac cellular damage produced by chemotherapeutic regimen.

Tanshinone IIA, a major component of Salvia milthorriza Bunge, inhibits platelet activation via Erk-2 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: The roots of Salvia milthorriza Bunge (Lamiaceae) known as "Danshen", are used in Traditional Chinese Medicine as a remedy for activating blood and eliminating stasis. TIIA, a diterpenoid of Salvia milthorriza, is one of active components in Danshen that exhibits a significant improvement of the blood flow in the coronary circulatory system and a reduction of myocardial infarction. However, its effect on platelet and underlying mechanism remains largely unknown. On this basis, this compound could be a promising agent to improve blood viscosity and microcirculation and to prevent CVD. MATERIALS AND METHODS: In order to investigate the effects of TIIA on platelet functionality and its interaction with various platelet activation pathways, rat PRP were incubated with TIIA for 1 min at 37°C prior the addition of the stimuli (ADP or collagen). Aggregation was monitored in a light transmission aggregometer measuring changes in turbidity with continuous observation up to 10 min after the addition of the stimuli. MAPK signaling pathway and tubulin acetylation were analyzed by a Western blot technique. The effect of the TIIA was also studied in vivo on bleeding time in mice. RESULTS: TIIA selectively inhibited rat platelet aggregation induced by reversible ADP stimuli (3 µM) in a concentration-dependent manner (0.5-50 µM). Nevertheless, TIIA was less active against the irreversible stimuli induced by ADP (10 µM) and collagen (10 µg/mL). Moreover, experiments performed on platelet lysates collected at different time-point after the addition of the stimuli shown that TIIA modulated tubulin acetylation and inhibited Erk-2 phosphorylation. Concomitantly, TIIA administrated i.p. at 10 mg/kg significantly amplified the mice bleeding time with an increase of 58% compared to its control (2.06±0.29 min vs 1.30±0.07). ASA was used as reference drug for in vitro and in vivo experiments. CONCLUSIONS: This study clarifies the intracellular signaling pathway involved in antiplatelet action of TIIA and also gives preliminary evidences for its anticoagulant activity. On this basis, this compound could be a promising agent to improve blood viscosity and microcirculation and to prevent CVD.

Use of Aspirin in normalization of recombinant human erythropoietin-mediated hyper-reactivity of platelets in rats.

OBJECTIVES: The cytokine erythropoietin is the primary stimulator of erythropoiesis and recombinant human erythropoietin (rHuEPO), which is widely used in the treatment of anemia associated with advanced chronic kidney disease (CKD). Adverse cardiovascular outcomes have been observed during clinical trials of anemia correction with rHuEPO in CKD patients. We investigated the effects of short-term, high-dose treatment with rHuEPO on platelet reactivity and effects of aspirin on platelet reactivity in healthy rats. MATERIALS AND METHODS: Animals received three daily dose of rHuEPO (25 µg/kg s.c.). Platelets were isolated after 48 h of last dose of rHuEPO to study the arachidonic acid-induced platelet aggregation. Aspirin (75 mg/kg p.o.) was given to animals just before 1 h of isolation of platelets. RESULTS: In rats, treatment with rHuEPO increased platelet reactivity and platelet count. The increased platelet reactivity was paralleled by decreased time-to-occlusion (TTO) in arterial thrombosis model, and decreased bleeding time after tail transection in rats. Treatment with rHuEPO followed by single dose of aspirin showed significant reduction in TTO and bleeding time as compared with aspirin-treated group. CONCLUSIONS: These findings suggest that rHuEPO increases platelet reactivity and aspirin normalizes the hyper-reactive platelet and may reduce the cardiovascular events associated with rHuEPO in CKD patients.