Anti-hemostatic, antithrombotic, and chemical profiles of a curly-leaf variety of Petroselinum crispum (Apiaceae), a food and medicinal aromatic herb.

Thrombosis is currently among the major causes of morbidity and mortality in the World. New prevention and therapy alternatives have been increasingly sought in medicinal plants. In this context, we have been investigating parsley, Petroselinum crispum (Mill.) Nym, an aromatic herb with two leaf varieties. We report here the in vitro, in vivo, and ex vivo anti-hemostatic and antithrombotic activities of a parsley curly-leaf variety. Aqueous extracts of aerial parts (PCC-AP), stems (PCC-S), and leaves (PCC-L) showed significant in vitro antiplatelet activity. PCC-AP extract exhibited the highest activity (IC50 2.92 mg/mL) when using ADP and collagen as agonists. All extracts also presented in vitro anticoagulant activity (APTT and PT) and anti-thrombogenic activity. PCC-S was the most active, with more significant interference in the factors of the intrinsic coagulation pathway. The oral administration of PCC-AP extract in rats caused a greater inhibitory activity in the deep vein thrombi (50%; 65 mg/kg) than in arterial thrombi formation (50%; 200 mg/kg), without cumulative effect after consecutive five-day administration. PCC-AP extract was safe in the induced bleeding time test. Its anti-aggregating profile was similar in ex vivo and in vitro conditions but was more effective in the extrinsic pathway when compared to in vitro results. Apiin and coumaric acid derivatives are the main compounds in PCC-AP according to the HPLC-DAD-ESI-MS/MS profile. We demonstrated for the first time that extracts from different parts of curly parsley have significant antiplatelet, anticoagulant, and antithrombotic activity without inducing hemorrhage, proving its potential as a source of antithrombotic compounds.

Antioxidant and hemostatic properties of preparations from Asteraceae family and their chemical composition - Comparative studies.

Asteraceae, known as sunflower family, is one of the largest flowering plants family around the world. Sunflower family contains numerous phytochemical compounds. The aim of this study was to describe phytochemical characteristics and investigate the effect of four sunflower vegetable preparations (extracts): chicory leaves (Cichorium intybus), green lettuce leaves (Lactuca sativa), red lettuce leaves (Lactuca sativa var. crispa) and sunchoke roots (Helianthus tuberosus) on different biomarkers of oxidative stress in human plasma in in vitro model. The antioxidant potential was also tested using the DPPH method. The phytochemical composition of the tested preparations was determined by UPLC-ESI-QTOF-MS. All the tested extracts demonstrated antioxidant activity in human plasma. We have observed chicory's and sunchoke's extracts had strongest antioxidant properties in the used models with human plasma. None of the tested vegetables changed ORAC and TAC in vitro. The obtained results suggest that sunflower vegetables might help to prevent oxidative stress related with cardiovascular diseases.

Comparisons of the anti-inflammatory, antiviral, and hemostatic activities and chemical profiles of raw and charred Schizonepetae Spica.

ETHNOPHARMACOLOGICAL RELEVANCE: A common view in traditional Chinese medicine (TCM) theory is that "processing can alter the efficacy of crude drugs". The clinical usage of some processed products may have already changed greatly over time during the development of modern scientific analysis. Therefore, the view of "processing can alter the efficacy of crude drugs" should be confirmed by comparative studies. Schizonepetae Spica (SS), a Chinese medicinal herb, is the dried spike of Schizonepeta tenuifolia Briq. It is available in two forms: raw products and charred products (Schizonepetae Spica Carbonisata, SSC; raw SS processed by stir-frying until carbonization). Raw SS is commonly used to treat TCM symptoms that resemble common cold, fever, respiratory tract infection and allergic dermatitis, while SSC has long been used as a remedy for TCM symptoms that resemble bloody stool and metrorrhagia. AIM OF THE STUDY: We aimed to examine whether stir-fry processing alters the anti-inflammatory, antiviral and hemostatic activities of SS and explore the chemical profile behind the potential changes in medicinal properties caused by stir-fry processing. MATERIALS AND METHODS: We used cell models to examine the anti-inflammatory and antiviral effects of raw SS and SSC. The bleeding time of the tail bleeding model and clotting time of the capillary method in mice were used to compare the hemostasis properties of raw SS and SSC. The chemical profiles of SS and SSC were compared using a method combining gas chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) analysis. RESULTS: The anti-inflammatory effects of SSC were less potent than those of raw SS. Both raw SS and SSC effectively inhibited viral infection in a dose-dependent manner, with IC50 values of 96.30 and 9.73 µg/mL and selectivity index (SI) values of >1.56 and 7.78, respectively. Interestingly, SSC showed more potent antiviral activities than raw SS. Intragastric administration of raw SS and SSC to mice demonstrated that the hemostatic effects of SSC were more potent than those of raw SS. By comparing the volatile chemical profiles of SSC, we found that twenty-nine constituents disappeared and that fifty-four new constituents were formed while the relative contents of five other components decreased and three other components increased. Additionally, the nonvolatile chemical profiles of raw SS and SSC differed, with thirty-two lower peaks and seven higher peaks in SSC than in SS. CONCLUSION: Our study showed that raw SS and SSC support traditional practice for the clinical applications of these two products except for raw SS used for the treatment of viral infection. It is a fascinating challenge to form SSCs with both traditional hemostatic activities and antiviral properties after stir-fry processing. In addition, the volatile and nonvolatile chemical constituents of raw SS changed dramatically during processing. Further studies are warranted to explore whether the change in chemical constituents is in accordance with the purpose of processing.

Anti-inflammatory and hemostatic effects of Linaria reflexa Desf.

The work presented here was aimed to investigate the in vivo anti-inflammatory and in vitro hemostatic activities of Linaria reflexa extract and to establish the relationship between its bioactivity and chemical composition. Twenty-three secondary metabolites were identified, most of them are good anti-inflammatory agents, in line with data by carrageenin-induced rat paw edema assays of the n-butanol extract showing high anti-inflammatory inhibition (63.90%) of edema swelling in the rat paw at the dose 200 mg/kg after 4 h. Furthermore, both extent of inflammatory response and tissue injury were prevented keeping the levels of rate myeloperoxidase (60.16%) and of malondialdehyde, which is the final product of lipid peroxidation generated by free radicals (58.58%). The same extract showed also a remarkable hemostatic effect established by measuring the coagulation time of decalcified plasma (45 s), related to its flavonoid glycosides content.

Evaluation of VEGF mediated pro-angiogenic and hemostatic effects and chemical marker investigation for Typhae Pollen and its processed product.

ETHNOPHARMACOLOGICAL RELEVANCE: Typhae Pollen (TP) is a well-known Traditional Chinese Medicine (TCM) to remove blood stasis. Carbonized Typhae Pollen (CTP), a processed product of TP after being stir-fried, has been widely applied to clinical practice with its capability of hemostasis. However, the underlying mechanism of TP and CTP are still not fully elucidated and discrimination against TP and CTP remains a challenge. AIM OF STUDY: The aim of this study is to investigate whether TP could remove blood stasis by promoting angiogenesis and the process of carbonizing it could enhance hemostatic effect. Meanwhile, some chemical markers for quality control of CTP had better to be found. MATERIAL AND METHODS: The changes of constituents between TP and CTP were analyzed by UPLC-QTOF-MS/MS. We investigated pro-angiogenic and hemostatic effects of TP and CTP in two zebrafish models: VRI-induced ISV insufficiency model and Ator-induced cerebral hemorrhage model. Subsequently, quantitative real-time PCR (qRT-PCR) was applied to investigate the mechanism of pharmacological effects. Finally, chemometric method was applied to find chemical markers. RESULTS: A total of 19 compounds were identified in qualitative analysis. The loss rate of each compound was calculated and compared. Two compounds (huaicarbon A/B) could only be detected in CTP and the content of flavonoid glycosides in CTP was significantly decreased compared with TP. The average content of the three identified flavonoid aglycones (quercetin, isorhamnetin and kaempferol) was increased about 30 percent in CTP. TP promoted pro-angiogenesis by up-regulating the expression of VEGFA, flt1 and kdr. After heating process, the pro-angiogenic activity was reduced and hemostatic activity was enhanced in CTP. Then qRT-PCR analysis found that CTP could significantly up-regulate the expression of VEGFA and vWF. In the discovery of markers, 6 chemical markers for discrimination of TP and CTP were obtained by chemometric method. CONCLUSION: Our research indicated that the pro-angiogenic activity of TP was involved in VEGF signaling pathway. After processing, hemostatic activity of CTP has been enhanced by up-regulating the expression of VEGFA and vWF. A chemical marker database was established to provide a scientific evidence for quality control, mechanism and the clinical application of TP and CTP.

Elucidation of the active ingredients of Lamiophlomis herba against hemorrhage based on network pharmacology and tail snipping model in mice.

This study aimed to elucidate the active ingredients of Lamiophlomis herba (LH), the overground part of Lamiophlomis rotata (Benth.) Kudo, against hemorrhage based on network pharmacology and tail snipping model in mice. A total of 118 hemorrhage-related target genes were identified by retrieving public databases, and 39 genes were identified as the hub genes of hemorrhage based on protein-protein interaction and module analyses. The interactions between 67 potentially active ingredients in LH and 7 genes in the 39 hub genes were established and analyzed through molecular docking and Cytoscape. A total of 21 ingredients were involved in the interactions, and were divided into three categories: iridoid (15 ingredients), flavonoid (2 ingredients) and other category (4 ingredients). Based on the "multi-ingredient, multi-target" characteristic of traditional Chinese medicines (TCMs), the results of network pharmacology indicated that iridoid might be the key active ingredient group of LH against hemorrhage. The contribution of iridoid to the hemostatic effect of LH was investigated by the tail snipping model in mice. The results showed that iridoid was the key active ingredient group of LH against hemorrhage, which confirmed the prediction in network pharmacology. Additionally, the previous reports also supported this prediction. In conclusion, the finding of the present study indicates that iridoid is the key hemostatic ingredient group of LH. This work provides valuable references for investigation of the hemostatic ingredients of LH based on the holistic theory of TCMs. Meanwhile, this work also provides further insight into the development of hemostatic drugs based on LH.

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.

Identification of certain Panax species to be potential substitutes for Panax notoginseng in hemostatic treatments.

Panax notoginseng (Burkill) F. H. Chen ex C. H. Chow (P. notoginseng) is a highly valued Chinese materia medica having a hemostatic effect and mainly used for the treatment of trauma and ischemic cardiovascular diseases. Stringent growth requirements, weak resistance to insect pests and plant diseases, arsenic contamination and continuous cropping constitute hurdles to further increases in the agricultural production of P. notoginseng. This review focuses on the traditional uses (based on traditional Chinese medicine theory), major chemical components, biological activities, pharmacological properties, geographical distributions and historical development of taxonomy of P. notoginseng and its related species in Panax genus, including Panax japonicus C. A. Meyer (P. japonicus), Panax japonicus C. A. Meyer var. major (Burkill) C. Y. Wu et K. M. Feng (P. japonicus var. major) and Panax japonicus C. A. Meyer var. bipinnatifidus (Seem.) C. Y. Wu et K. M. Feng (P. japonicus var. bipinnatifidus) are reviewed. This review sheds light on the origin herbs of Zhujieshen (ZJS) and Zhuzishen (ZZS), e.g., P. japonicas var japonicas, P. japonicus var. major and P. japonicus var. bipinnatifidus could be used as a substitute for P. notoginseng as hemostatic herbs.

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.

Evaluation of Artemisia scoparia for hemostasis promotion activity.

Excessive hemorrhage through any reasons is a life threatening process. Artemisia scoparia of family Asteraceae has been used in local system of medicine to stop bleeding from wounds and in injuries, antiseptic, in healing urticarial and for removal of worms from the body. Aerial parts of A. scoparia was extracted with 95% methanol (ASM) and fractionated through liquid-liquid partition in ascending order of n-hexane (ASH), chloroform (ASC), ethyl acetate (ASE), and the remaining as the aqueous fraction (ASA). Phytochemical classes of the extract/fractions were determined by qualitative assays. Prothrombin time (PT) was estimated on the plasma of human blood by Owren method. Capillary tube method was applied to determine the hemostasis activity in Sprague-Dawley rat. Tannins, saponins, terpenoids, quinones, betacyanins and flavonoids were present whereas phlobatannins, anthraquinones and alkaloids were established absent in ASM, ASC, ASE and ASA. Prothrombin time was significantly decreased by mixing (10 µg) of ASM (16.67±1.15 sec), ASH (12.33±0.57 sec), ASC (15.33±0.57 sec) and ASA (9.0±1.0 sec) to that of vehicle (20.0±1.0 sec). Administration (200 mg/kg) of all the extract/fractions showed significantly less (26.00±11.79 sec - 41.00±7.21 sec) hemostasis time as compared to the (242.67±39.67 sec) control rats. The results suggested the therapeutics importance of A. scoparia use in bleeding pathologies.

Isolation, Characterization and Evaluation of Collagen from Jellyfish Rhopilema esculentum Kishinouye for Use in Hemostatic Applications.

Hemostat has been a crucial focus since human body is unable to control massive blood loss, and collagen proves to be an effective hemostat in previous studies. In this study, collagen was isolated from the mesoglea of jellyfish Rhopilema esculentum Kishinouye and its hemostatic property was studied. The yields of acid-soluble collagen (ASC) and pepsin-soluble (PSC) were 0.12% and 0.28% respectively. The SDS-PAGE patterns indicated that the collagen extracted from jellyfish mesoglea was type I collagen. The lyophilized jellyfish collagen sponges were cross-linked with EDC and interconnected networks in the sponges were revealed by scanning electron microscope (SEM). Collagen sponges exhibited higher water absorption rates than medical gauze and EDC/NHS cross-linking method could improve the stability of the collagen sponges. Compared with medical gauze groups, the blood clotting indexes (BCIs) of collagen sponges were significantly decreased (P < 0.05) and the concentration of collagen also had an influence on the hemostatic property (P < 0.05). Collagen sponges had an improved hemostatic ability compared to the gauze control in tail amputation rat models. Hemostatic mechanism studies showed that hemocytes and platelets could adhere and aggregate on the surface of collagen sponge. All properties make jellyfish collagen sponge to be a suitable candidate used as hemostatic material and for wound healing applications.

Exploring hemostatic and thrombolytic potential of heynein - A cysteine protease from Ervatamia heyneana latex.

ETHNOPHARMACOLOGICAL RELEVANCE: The latex of Ervatamia heyneana (Wall.) T. Cooke plant has been used for wound healing and various skin diseases by Indian tribes and folklore. AIM OF THE STUDY: To validate the scientific basis of heynein - a key protease of Ervatamia heyneana, in hemostasis and wound healing process. MATERIALS AND METHODS: The latex from E. heyneana was processed and subjected to two step purification. The purified heynein was assayed for proteolytic activity using casein as substrate and also attested by zymography. The inhibition studies confirmed the nature of heynein. Pure fibrinogen was used for fibrinogenolytic activity and citrated plasma was used for coagulant and fibrinolytic activities. The edema inducing action and hemorrhagic activity of heynein were assessed on mice model. RESULTS: The purified heynein exhibited proteolytic activity, which was confirmed by caseinolytic assay and zymography. The inhibition studies confirmed heynein to be a cysteine protease. Heynein showed complete hydrolysis of all the three subunits of human fibrinogen (Aα, Bß, γ). It exhibited strong pro-coagulant activity by reducing plasma clotting time from 248 to 39s at 40µg concentration. Heynein cleaved α polymer subunit in fibrin clot and did not induce edema and hemorrhage in mice models. The non-hemorrhagic nature was supported with histopathological studies of skin samples. CONCLUSION: Heynein displays strong pro-coagulant action associated with fibrin(ogen)olytic activity. This provides basis for the observed pharmacological action of Ervatamia heyneana and thereby justifies its use in folk medicine.

Bletilla striata: Medicinal uses, phytochemistry and pharmacological activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Bletilla striata (Thunb.) Reichb. f. (Orchidaceae), also known as Hyacinth Orchid and Baiji (Simplified Chinese: ), not only has been widely used for the treatment of hematemesis, hemoptysis, and traumatic bleeding due to the efficacy of arresting bleeding with astringent action, but also has been applied topically to overcome ulcers, sores, swellings, and chapped skin due to the efficacy of dispersing swelling and promoting tissue regeneration. Additional medical applications include the treatment of tuberculosis, malignant ulcers, hemorrhoids, anthrax, eye diseases, and silicosis. AIM OF THIS REVIEW: This review aims to provide up-to-date information on the botanical characterization, medicinal uses, chemical constituents, pharmacological activities, and toxicity of B. striata. In addition, this paper also focuses on the possible exploitation of this plant for the treatment of different diseases, and uncovers opportunities for future research. MATERIALS AND METHODS: The relevant information on B. striata was gathered from worldwide accepted scientific databases via an electronic search (Google Scholar, Web of Science, ScienceDirect, ACS Publications, PubMed, Wiley Online Library, SciFinder, CNKI). Information was also obtained from The Plant List, Chinese pharmacopoeia, Chinese herbal classics books, PhD and MSc dissertations, etc. RESULTS: A comprehensive analysis of the literature obtained through the above-mentioned sources confirmed that the ethnomedical usages of B. striata have been recorded in Mongolia, Korea, Japan, and China. Phytochemical investigations revealed that the major chemical constituents of B. striata are polysaccharides, bibenzyls, phenanthrenes, triterpenoids and its saponins, steroids and its saponins, which also have been proven to be the main bioactive substances capable of exhibiting numerous pharmacological activities including wound healing, antiulcer, hemostasis, cytotoxicity, antimicrobial, anti-inflammation, anti-oxidation, immunomodulation, anti-fibrosis, antiaging, anti-allergy, and anti-itch. CONCLUSIONS: Preliminary investigations on pharmacological properties of B. striata have shown that B. striata is an outstanding astringent hemostatic medicinal, B. striata polysaccharides (BSP) as the major bioactive components not only capable of promoting wound healing, but also show good performance as a kind of promising natural biomaterial. More importantly, BSP are also reported to be excellent embolic material. However, further investigations need to be carried out to fully clarify its efficacy of dispersing swelling and promoting tissue regeneration. Moreover, this plant also needs a lot more investigations to clarify the pathways of absorption, distribution, metabolism and excretion, and to evaluate its long-term in vivo chronic toxicity before proceeding to the development of pharmaceutical formulation.

Cytotoxic, anti-inflammatory and hemostatic spirostane-steroidal saponins from the ethanol extract of the roots of Bletilla striata.

Four new spirostane steroidal saponins, (1α,3α)-1-O-[(ß-d-xylopyranosyl-(1→2)-α-l-rhamnopyranosyl)]-3-O-d-glucopyranosyl-5α-spirostan (1), (1α,3α)-1-O-[(ß-d-xylopyranosyl-(1→2)-α-l-rhamnopyranosyl)oxy]-3-O-d-glucopyranosyl-25(27)-ene-5α-spirostan (2), (1α,3α)-1-O-[(ß-d-xylopyranosyl-(1→2)-α-l-rhamnopyranosyl)oxy]-epiruscogenin (3), and (1α,3α)-1-O-[(ß-d-xylopyranosyl-(1→2)-α-l-rhamnopyranosyl)oxy]-epineoruscogenin (4) together with two known compounds, bletilnoside A (5) and 3-O-ß-d-glucopyranosyl-3-epi-neoruscogenin (6), were isolated from the ethanol extract of the roots of Bletilla striata (Thunb.) Reichb. f. The structures of the isolated compounds were established based on 1D and 2D ((1)H-(1)H COSY, HMQC, and HMBC) NMR spectroscopy, in addition to high resolution mass spectrometry. The isolated compounds were tested in vitro for cytotoxicities against seven tumor cell lines, anti-inflammatory activities against Cox-1 and Cox-2, and hemostatic activities. As a result, compounds 1-4 and 6 exhibited significant cytotoxicities against all the tested tumor cell lines with IC50 value less than 30µM and selective inhibition of Cox-2 comparable with the standard drug NS-398 (>90%). Additionally, compounds 1-6 showed potent hemostatic activities.

[Filtration of active fractions with hemostasis effect from platycladi cacumen carbonisatum].

OBJECTIVE: To discuss the hemostasis of the different polarities of Platycladi Cacumen Carbonisatum (PCC) on the blood heat and hemorrhage syndrome rat model induced by dry yeast. METHOD: The SD rats were divided into seven groups. Yunnan Baiyao was taken as the positive control drug. The rats in the control group and model group were fed with CMC-Na for 7 days, and the rats in other groups were fed with corresponding drugs simultaneously. On day 7, the blood heat and hemorrhage syndrome rat model was established. Indexes including the whole blood viscosity, plasma viscosity, thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen content (FIB), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), blood platelet count (PLT), thrombocytocrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW) and the rate of platelet aggregation induced by ADP were detected. Additionally, the pathological examinations of lungs among each group were compared. RESULT: Compared with the control group, the RBC, HGB and HCT of rats in the model group increased significantly, with distinct increase in high, middle and low whole blood viscosity and plasma viscosity of rats in the model group; TT and APTT were notably prolonged, while PT was notably shortened, with significant increase in FIB content; PLT, PCT, MPV and PDW remarkably increased; Additionally, the rate of platelet aggregation induced by ADP significantly decreased. After ig administration of the ethyl acetate extract of PCC, the low whole blood viscosity and plasma viscosity remarkably decreased; TT and APTT were significantly shortened, with notable reduction in PDW and in FIB content Additionally, the rate of platelet aggregation induced by ADP significantly increased. The injury of lungs was also improved in ethyl acetate extract group. The rate of platelet aggregation induced by ADP of n-butanol extract group notablly increased. Plasma viscosity of water extract group remarkably decreased, with TT being significantly shortened. But the effects of n-butanol extract or water extract were weaker than that of ethyl acetate extract. And the effect of petroleum ether extract was the weakest. CONCLUSION: Ethyl acetate extract is the active part of PCC, showing the effect of hemostasis by reducing the low whole blood and plasma viscosity, improving coagulation function mainly by acting on the endogenous coagulation, and ameliorating the function of platelet aggregation.

Chemical constituents from Sedum aizoon and their hemostatic activity.

CONTEXT: Sedum aizoon L. (Crassulaceae) (SA) is widely used to treat various hemorrhages in folk medicine. However, its hemostatic constituents are not yet clear. OBJECTIVE: The chemical constituents of EtOAc fraction from SA and their hemostatic activity were investigated to provide a basis for the application in folk use. MATERIALS AND METHODS: The chemical constituents were isolated from the aerial parts of SA by column chromatography and identified by IR, MS, and NMR, then tested for hemostatic activity using the capillary method and coagulation assays including blood clotting time in vivo, and prothrombin time (PT), activated partial thromboplastin time (APTT), and thrombin time (TT) in vitro at concentrations of 300.0, 100.0, and 30.0 µg/mL. RESULTS: Eleven compounds were identified as p-hydroxybenzoic acid (1), gallic acid (2), protocatechuic acid (3), vallinic acid (4), thymine (5), caffeic acid (6), 5,7-dihydroxy chromone (7), pyrogallol (8), quercetin (9), kaempferol (10), and luteolin (11). This is the first report of compounds 3-8 being isolated from this plant. Compounds 2 (300.0 and 100.0 µg/mL), 4 (100.0 µg/mL), and 11 (100.0 and 30.0 µg/mL) significantly reduced the clotting time (p < 0.01) with inhibition rates of 34.7, 24.5, 30.3, 25.9, and 36.6%, respectively. For further mechanism study, they also reduced PT (3.5, 2.5, 3.5, 3.5, and 3.8%, respectively), APTT (4.5, 3.3, 11.4, 8.5, and 11.1%, respectively), and TT (20.3, 3.8, 7.6, 6.1, and 10.3%, respectively). DISCUSSION AND CONCLUSION: SA produced hemostatic activity possibly related to the presence of gallic acid, vallinic acid, and luteolin, which may be potent candidates of hemostatic drug.

A comparison of the ability of Bellucia dichotoma Cogn. (Melastomataceae) extract to inhibit the local effects of Bothrops atrox venom when pre-incubated and when used according to traditional methods.

Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A2 activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A2 activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd was administered in combination with BA. The SDS-PAGE profiles showed that several of the BaV protein (SDS-PAGE) and enzyme (zymography) bands were not detected when the venom was pre-incubated, and Western blot revealed that this was not caused by the AEBd enzymes observed in the zymogram. The "pseudo inhibition" observed after pre-incubation in this study may be due to the presence of tannins in the extract, which could act as chelating agents, removing metalloproteins and Ca²âº ions and thus inhibiting hemorrhagin and PLA2 activity. However, when administered according to traditional methods, B. dichotoma extract was effective in blocking BaV-induced edematogenic activity and had an additional effect on inhibition of this activity by BA.

Study on hemostatic activities of the rhizome of Paris bashanensis.

UNLABELLED: CONTEXT. Paris bashanensis Wang et Tang (Liliaceae) is widely used in traditional Chinese medicine for the treatment of injuries, fractures and hemorrhage in Hubei and Sichuan Province. OBJECTIVE: The n-BuOH extract of Paris bashanensis was investigated for hemostatic activity and chemical constituents in order to provide a basis for the application in folk use. MATERIALS AND METHODS: The n-BuOH extract of P. bashanensis was divided into three eluents (30, 50 and 70% EtOH) by macroporous adsorptive resin D101. The bleeding time of breaking tail hemostasis and clotting time of capillary and slide method in mice were used extensively to screen the hemostasis properties after repetitive administration of these three fractions (100 mg/kg, 50 mg/kg) for 5 days (total of 5 times, once per day). The chemical compounds were analyzed by HPLC-UV. RESULTS: The inhibition rates in the bleeding time of 70, 50 and 30% n-BuOH ext. were 45, 32 and 21%, respectively. Using the slide method the decreasing rate of the clotting time of 70, 50 and 30% n-BuOH ext. were 71, 65 and 32% and in the experiment of capillary method, the inhibition rates were 43, 31 and 24%, respectively. A total of 70% n-BuOH ext. showed a high content of the pennogenin-type saponins by HPLC-UV. DISCUSSION AND CONCLUSIONS: The 70% n-BuOH ext. of P. bashanensis was found to contain high levels of pennogenin saponins, which may lead to a higher hemostatic activity. Combined with the hemostatic test, P. bashanensis could be used as a resource of hemostatic drug.

[Pharmacological study on hemostasis, analgesic and anti inflammation effects of the alcohol extract of Hibiscus tiliaceus].

OBJECTIVE: To study the effects of hemostasis, analgesic and anti inflammation of the alcohol extract of Hibiscus tiliaceus and offer pharmacological and experimental basis for its safe and effective use in clinic. METHODS: The effects of hemostasist were observed with tail breaking method, capillary tube method and slide method; Hot board and body distortion induced by acetic acid methods were applied in mice analgesia experiment, the mice model of acute auricle swelling induced by dmi ethylbenzene and capillary permeability induced by acetic acid were applied to observe the anti inflammatory effects. RESULTS: The alcohol extract of Hibiscus tiliaceus could significantly reduce the bleeding time and the clotting time, delay the plant reaction time and reduce the writhing times of the mice, and it also had effect on inhibiting swelling of mice ear and the permeability of the capillary. CONCLUSION: These results suggest that the alcohol extract of Hibiscus tiliaceus has the effects of hemostasis, analgesic and anti inflammation.

In vivo and in vitro hemostatic activity of Chromolaena odorata leaf extract.

CONTEXT: Chromolaena odorata (L.) R.M.King & H.Rob. (Asteraceae) or Siam weed has long been used to stop bleeding in Thailand and many countries. Only the aqueous leaf extract was investigated in in vivo and there have been conflicting results of in vitro hemostatic mechanisms of this plant. OBJECTIVE: The most appropriate C. odorata leaf extract that promoted the highest hemostatic activity and the hemostatic mechanisms of these plant extracts will be investigated. MATERIALS AND METHODS: The lyophilized aqueous leaf extract and alcoholic (50, 70, and 95% ethanol) extracts from the fresh and dried leaves were investigated both in vivo and in vitro. The bleeding time in male Wistar rats was measured to investigate the hemostatic effect. The hemostatic mechanisms were tested using in vitro platelet aggregation and blood coagulation tests in sheep plasma. RESULTS: All extracts displayed significantly reducing bleeding time (<2.5 min) in rats but did not induce platelet aggregation or blood clotting in the in vitro study. The in vitro blood clotting times of all extracts were >0.6 min. Ethanol extract (70%) from the dried leaves proved to be the extract producing the highest hemostatic activity in vivo with the bleeding time of 1.85 min. DISCUSSION AND CONCLUSION: The in vivo study with rats confirmed the significant ability of this plant extract to stop bleeding. However, the sufficient amount of calcium and active compounds which are aggregating and clotting agents to enhance blood coagulation and platelet aggregation in in vitro tests should be further studied.