Thrombolytic Activity Research Articles

Multifaceted Chemico-Pharmacological Insights into Cynometra ramiflora L.: Unveiling its GC-MS, Cytotoxic, Thrombolytic, Anti-Inflammatory, Antioxidant, Anti-Diarrheal, Hypoglycemic, and Analgesic Potentials

Objective: This study aimed to comprehensively evaluate the chemico-pharmacological properties of methanolic extractives derived from Cyanometra ramiflora leaves, encompassing a spectrum of activities including cytotoxic, thrombolytic, membrane-stabilizing, antioxidant, antidiarrheal, hypoglycemic, and analgesic effects. Methods: Phytomolecules were identified using Gas Chromatography-Mass Spectrometry (GC-MS), while antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total phenolic content tests. Cytotoxicity was determined through the brine shrimp bioassay, while thrombolytic activity was evaluated via clot lysis assays, and membrane-stabilizing effects were investigated for potential anti-inflammatory properties. In-vivo experiments using mice were conducted solely with the methanolic extract to assess central and peripheral analgesic, hypoglycemic, and antidiarrheal properties. Results: The presence of 47 compounds in the methanolic extract, with tetratriacontyl pentafluoropropionate (9.25%), pentatriacontane (8.75%), and 2-methylhexacosane (5.9%) being predominant. The dichloromethane-soluble fraction exhibited the highest cytotoxicity, with a 50% lethal concentration (LC50) of 11.05 µg/mL. Both petroleum-ether and methanol-soluble fractions demonstrated more than 50% lysis in thrombolytic activity assays. Under hypotonic and heat-induced conditions, the ethyl acetate and dichloromethane-soluble fractions of C. ramiflora significantly reduced erythrocyte hemolysis. Notably, the methanol-soluble fraction exhibited significant antioxidant potential with 50% inhibitory concentration (IC50) of 17.71 μg/mL compared to the standard EDTA (IC50 = 75.94 μg/mL). The hypoglycemic investigation revealed a 63.73% reduction at 90 min with a 600 mg/kg dose, while mild antidiarrheal activity was observed with a 37.5% reduction in diarrhea at 4 h intervals. Significant analgesic effects were demonstrated in both central and peripheral tests, with a 68.53% reduction in writhing during the peripheral test and an average tail immersion time of 5.38 s at 90 min. Conclusion: C. ramiflora leaves show promise as natural sources for various pharmacological activities, including cytotoxicity, thrombosis, inflammation, antioxidant, hypoglycemic, and analgesic effects. However, further comprehensive research is recommended to elucidate the precise mechanisms of action and analyze the phytochemical composition of C. ramiflora plants.

Abstract 148: Loss Of Tlr4 Signaling Inhibits Thrombus Resolution In A Non-monocyte Dependent Manner

Background: Loss of TLR4 signaling reduces Venous Thrombosis (VT) resolution. Leukocyte expression of TLR4 is elevated in humans undergoing acute to chronic DVT transformation. We hypothesized that TLR4 signaling in monocytes directs thrombus clearance. Methods: Wild type, Tlr4 -/- , LysM cre+/- Tlr4 fl/fl mice underwent VT via IVC stasis at 4/7/14d timepoints. Thrombi subtypes were identified by flow cytometry. Thrombus size was assessed, and fibrinolytic mediators were assayed in splenic/bone marrow Mφs. Whole blood from mice was stimulated with TLR2/4 and assayed for thrombolytic potential via HALO assay. Immunohistochemistry was performed for cellular infiltrate. Results: We observed elevated TLR4 cell surface expression on CD11b+/Ly6C- Mo/Mφ from wild type thrombi. Thrombus weight in Tlr4 -/ mice was elevated at 4d stasis, confirming impaired thrombus clearance. In basal conditions, circulating Mo/Mφ saw decreased Urokinase & MMP9 mRNA levels in LysM cre+/- mice. In post thrombotic conditions, we saw decreased protein levels of Urokinase and MMP9 in Cre+ bone marrow-derived Mφs. Whole blood thrombolysis efficiency increased with TLR4 agonism in Cre- mice compared to Cre+. Efficiency was restored with LPS-mediated dual TLR2/4 agonism. Total thrombus weight was unchanged. No difference was identified in monocyte infiltrate into the thrombus (Fig. 1) Conclusions: Genetic deficiency of Mo/Mφ TLR4 signaling alters induction of fibrinolytic/matrix remodeling gene expression. Ex vivo TLR4 agonism increased whole blood thrombolysis in Cre- but not Cre+ mice. Additional TLR2 agonism of Cre+ mice restored thrombolytic activity, suggesting TLR2 as a compensatory pathway that may account for the unchanged in vivo phenotype in LysMcreTlr4fl/fl mice. Further clarifying these cellular pathways may provide an alternate pathway to target VT resolution.

Evaluation of In Vitro Antimicrobial, Cytotoxic, Thrombolytic, and Antiarthritic Property of Different Parts of Bari Orchid.

Many different herbal extracts have historically been utilized to treat microbe-induced infections, injuries, cancer, thrombosis, and arthritis. The purpose of this study was to determine the antibacterial, cytotoxic, in vitro thrombolytic, and in vitro antiarthritic properties of ethanolic extracts of stem and seed of Bari orchid 1 (BO) plant. This orchid plant was developed by the Bangladesh Agriculture Research Institute (BARI) in Gazipur. Fourteen microbes were employed in the antimicrobial investigation, and samples of orchids were compared to ciprofloxacin as a reference. The BO/seed extract was found to possess more antibacterial activity. The lethality test of brine shrimps was used to assess the LC50 values. The BO/stem extract exhibited a higher cytotoxicity potential, in comparison to the BO/seed extract. Two concentrations (1000 and 100 ppm) and two incubation times (24 hours and 1.5 hours) were used to assess the thrombolytic activity of the extracts. Regarding the thrombolytic effect, the BO/stem extract has demonstrated greater promise. Furthermore, the herbal extract's antiarthritic activity was investigated at four different concentrations, and the results were evaluated in comparison with those of diclofenac sodium. When comparing BO/stem extract to other extracts, the greatest values for protein denaturation were obtained.

Targeted Thrombolysis with Magnetic Nanotherapeutics: A Translational Assessment.

Plasminogen activators, such as recombinant tissue-type plasminogen activators (rtPAs), while effective in treating thromboembolic diseases, often induce hemorrhagic complications due to non-specific enzyme activities in the systemic circulation. This study evaluated the targeting efficiency, efficacy, biodistribution, and potential toxicity of a rtPA covalently attached to chitosan-coated magnetic nanoparticles (chitosan-MNP-rtPA). The thrombolytic activity of a chitosan-MNP-rtPA was preserved by protection from an endogenous plasminogen activator inhibitor-1 (PAI-1) in whole blood and after circulation in vivo, as examined by thromboelastometry. Single-photon emission computed tomography (SPECT) demonstrated real-time retention of a 99mTc-MNP-rtPA induced by magnet application in a rat embolic model; an 80% reduction in rtPA dosage for a chitosan-MNP-rtPA with magnetic guidance was shown to restore blood flow. After treatment, iron deposition was observed in the reticuloendothelial systems, with portal edema and neutrophil infiltration in the liver at a ten-fold higher dose but not the regular dose. Nevertheless, no liver or renal toxicity was observed at this higher dose. In conclusion, the liver may still be the major deposit site of rtPA nanocomposites after targeted delivery; chitosan-coated MNPs are potentially amenable to target therapeutics with parenteral administration.

In vivo Anti-Inflammatory and Antinociceptive Activity Evaluation of Brassica Rapa Ssp. Chinensis Ethanolic Extract with In Vitro Thrombolytic and Anthelmintic Activity Test

Objective: Brassica rapa ssp. chinensis is a well-known vegetable plant (Family: Brassicaceae) often grown as a type of Chinese cabbage. The present study was designed to explore the thrombolytic, anthelmintic, anti-inflammatory, and antinociceptive activities of ethanolic extract of the Brassica rapa ssp. chinensis leaves. Methods: The in vitro thrombolytic activity of the leaf extract was evaluated by clot analysis using Streptokinase as a standard drug. The in vitro anthelmintic activity was assessed by applying four different concentrations of the plant extract (10 mg/ml, 20 mg/ml, 40 mg/ml, and 60mg/ml) and recording the time of paralysis and death of worms. Two different doses (100mg/kg and 250mg/kg b.w.) were administered to evaluate the anti-inflammatory activity of the carrageenan-induced paw edema model. The antinociceptive activity was evaluated by hot plate test in mice by two different doses (200mg/kg and 400 mg/kg b.w.). Results: The ethanolic leave extract had thrombolytic action, which broke up blood clots that had already formed. The ethanolic leaf extract showed both paralysis and faster worm death at those concentrations in the anthelmintic test. The extract showed significant anti-inflammatory action in pedal edema compared to the control group. The results show that the highest level of anti-inflammatory action against carrageenan-induced hind paw edema may be achieved with an ethanolic extract of Brassica rapa ssp. chinensis (250 mg/kg, p.o.). The proportion of maximum possible effect in the hot plate test was 82.49 percent in the antinociceptive evaluation. Conclusion: The pharmacological test of leaf extract endorsed the clinical uses of Brassica rapa ssp. chinensis in treating thrombosis, intestinal worms, and inflammation. Thus, this plant could be subjected to extensive bioactive compounds to discover novel therapeutic agents. Keywords: Brassica rapa ssp. chinensis, ethanolic extract, thrombolytic activity, anthelmintic activity, anti-inflammatory activity, antinociceptive activity

A comparative study on biosynthesized silver nanoparticles from H. undatus fruit peel and their therapeutic applications

The green synthesis of nanoparticles (NPs) gained significant impacts in various fields due to the use of eco-friendly approaches. In this study, silver nanoparticles (AgNPs) were synthesized from the aqueous extract of Hylocereus undatus fruit peel. The presence of AgNPs was analysed using characterization methods such as UV‒Vis, FTIR, GCMS, XRD, EDAX, and FESEM. The synthesized AgNPs showed greater antibacterial activity against Escherichia coli than against Streptococcus pneumoniae. The antifungal activity against Candida albicans was greater than that against Candida tropicalis. The IC50 value for the antibiofilm activity of the AgNPs was 2.81 µg/mL, whereas that of the H. undatus peel extract was 1.34 µg/mL. The invitro antioxidant activity of the AgNPs was evaluated using two different methods. The DPPH radical scavenging activity of the AgNPs and fruit peel extract was observed with IC50 values of 3.8 and 2.03 µg/mL respectively. On the other hand, nitric oxide radical scavenging activities were recorded and the IC50 values were calculated to be 2.8 and 2.3 µg/mL. The AgNPs demonstrated thrombolytic activity in human blood with 10, 32.36, and 56.25% lysis. The cytotoxicity of the AgNPs was minimal, with an IC50 of 0.2 µg/mL and the peel extract had the greatest cytotoxicity with an IC50 of 0.3 µg/mL. The findings of this study demonstrated that the synthesized AgNPs from H. undatus peel extract could be potential candidates for treating prostate cancer.Graphical abstract

Ascorbic acid-mediated selenium nanoparticles as potential antihyperuricemic, antioxidant, anticoagulant, and thrombolytic agents

Abstract Selenium (Se) is an important trace element that is involved in controlling oxidative stress and inflammatory disorders. Gouty arthritis is the inflammation and pain within the joints and tissues caused due to the accumulation of monosodium urate (MSU) crystals. This study aimed to investigate the antigout, antioxidant, anticoagulant, and thrombolytic potential of ascorbic acid-mediated Se nanoparticles (A-SeNPs). Different analytical techniques were used to investigate the formation of A-SeNPs. The antigout potential of the nanoparticles was carried out using MSU crystal dissolution, uric acid (UA) degradation assay, and xanthine oxidase inhibition (XOI). A-SeNPs exhibited excellent antihyperurecemic activity in a concentration-dependent manner. It was observed that at the tested concentration of 20 mg·mL−1, the A-SeNPs demonstrated significant breakage and dissolution of MSU crystals and resulted in UA degradation of 67.76%. Similarly, A-SeNPs resulted in 76% XOI with an excellent IC50 of 140 µg·mL−1. Furthermore, considerable antioxidant activity was noted for the A-SeNPs as evaluated with multiple antioxidant assays. Finally, the NPs were found to have significant anticoagulant and thrombolytic potential. Thus, it was concluded that A-SeNPs have potent antihyperuricemic, antioxidant, anticoagulant, and thrombolytic activities, making them an ideal choice for future biomedical applications.

Chemico-pharmacological evaluation of the methanolic leaf extract of Catharanthus ovalis: GC-MS/MS, in vivo, in vitro, and in silico approaches.

Introduction: Natural plant-based medicines have gained popularity recently as a major source of inventive, risk-free, and more potent secondary bioactive compounds with medicinal potential. Catharanthus ovalis is a perennial shrub containing various indole alkaloids cultivated extensively for local medical purposes. Methods: This research is conducted to identify the phytocompounds present in the leaves of C. ovalis and its central and peripheral analgesic, thrombolytic, and membrane-stabilizing activities through tail immersion, acetic acid-induced writhing, human blood clot lysis, and erythrocyte lysis by heat and hypotonic solution methods, respectively. Results and discussion: A total of 39 compounds were identified using GC-MS/MS techniques, including hexadecanoic acid, methyl ester (56.749%), methyl stearate (29.782%), carvacrol and its TBDMS derivative (12.586%), and 9-octadecenoic acid, methyl ester, (E)-] (9.297%) presented in high quantity. The highest tail immersion latency was observed for the 600mg/kg extract of C. ovalis crude extract. Both 400 and 600mg/kg doses of C. ovalis crude extract exhibited prominent peripheral analgesic activity. The maximum thrombolytic effect was observed by DCM soluble fraction extract by inhibiting 54.87% of the clot. However, the aqueous-soluble fraction of this extract manifested an excellent membrane-stabilizing effect by showing 73.98% and 87.51% hemolysis against heat- and hypotonic-induced hemolysis, respectively. Some of the compounds were identified as active agents against different receptors related to these diseases, which supported the findings of in vitro and in vivo tests. Conclusion: Further investigation needs to be conducted to specify and identify the exact mechanism of action of these compounds.

Modulation of Abundance and Location of High-Mobility Group Box 1 in Human Microglia and Macrophages under Oxygen-Glucose Deprivation.

While stroke represents one of the main causes of death worldwide, available effective drug treatment options remain limited to classic thrombolysis with recombinant tissue plasminogen activator (rtPA) for arterial-clot occlusion. Following stroke, multiple pathways become engaged in producing a vicious proinflammatory cycle through the release of damage-associated molecular patterns (DAMPs) such as high-mobility group box 1 (HMGB1) and heat shock protein 70 kDa (HSP72). HMGB1, in particular, can activate proinflammatory cytokine production when acetylated (AcHMGB1), a form that prefers cytosolic localization and extracellular release. This study aimed at determining how HMGB1 and HSP72 are modulated and affected following treatment with the anti-inflammatory compound resveratrol and novel platelet membrane-derived nanocarriers loaded with rtPA (CSM@rtPA) recently developed by our group for ischemic artery recanalization. Under ischemic conditions of oxygen-glucose deprivation (OGD), nuclear abundance of HMGB1 and AcHMGB1 in microglia and macrophages decreased, whereas treatment with CSM@rtPA did not alter nuclear or cytosolic abundance. Resveratrol treatment markedly increased the cytosolic abundance of HSP72 in microglia. Using proximity ligation assays, we determined that HSP72 interacted with HMGB1 and with acetylated HMGB1. The interaction was differentially affected under the OGD conditions. Resveratrol treatment under the OGD further decreased HSP72-HMGB1 interactions, whereas, in contrast, treatment increased HSP72-AcHMGB1 interactions in microglia. This study points out a salient molecular interaction suited for a two-pronged nanotherapeutic intervention in stroke: enhancement of rtPA's thrombolytic activity and modulation of cytosolic interactions between HMGB1 and HSP72 by resveratrol.

Evaluation of antimicrobial, thrombolytic and cytotoxic activities of ethanol flower extract of Bauhinia acuminata

Background: To develop the herbal drug with the least side effects, there are superior opportunities to discover the medicinal and other biological properties. Natural products serve as sources of beneficial chemical molecules. For this study, Bauhinia acuminate (Family: Fabaceae) having some pharmacological activities was chosen. The present studies was designed for the antimicrobial, thrombolytic and cytotoxic activity of ethanolic flowers extract of Bauhinia acuminata. Methods: The antimicrobial effect of the extracts was evaluated by Disc-Diffusion Assay Method and the cytotoxic activity was determined by brine shrimp lethality test. The thrombolytic activities of the plant extracts were evaluated by a method using streptokinase as a reference standard. Results: The extract has mild antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, enteropathegenic Escherichia coli, Salmonella typhi, Vibrio mimicus. The zone of inhibition of the extract was measured comparing with standard Tetracycline. Thrombolytic activity of flowers of Bauhinia acuminata showed mild (14.57%) clot lysis while the standard streptokinase showed (90%). Having moderate cytotoxic activities, the mortality rate of brine shrimp was observed in increasing order with the increasing of concentration of extract. In the present study of the ethanol extract LC50=3.251 µg/ml and LC90=286.57 µg/ml. However, Standard control LC50=0.0036 µg/ml and LC90=0.75 µg/ml were found respectively. Conclusions: Through the studies, it can be concluded that the ethanolic extract of Bauhinia acuminata could be used in drug formulation or pharmaceutical application. Hence, the plant may further be explored for its various pharmacological activities.

Toxic or medicinal? New approach to the pharmacological and toxicological properties of Echinodorus macrophyllus leaves

Echinodorus macrophyllus contains some phytochemicals that can induce toxicity but are also promising as ingredients in products to promote and maintain health, improve the immune system, and prevent diseases. In this study, the aqueous and ethanolic extracts of leaves of this species were characterized by high-performance liquid chromatography, investigated in vitro regarding their modulating action on snake venom enzymes, and evaluated in digestive enzyme inhibition assays. The extracts inhibited the phospholipase activity induced by Bothrops jararacussu and B. moojeni venoms. The hemolytic activity induced by Crotalus durissus terrificus venom was potentiated by 30-73% by the ethanolic extract at the ratios analyzed. The thrombolytic activity induced by B. moojeni venom was inhibited by 51% and 34% by the aqueous and ethanolic extracts at a ratio of 1:5 (w:w), respectively. The clotting time induced by B. moojeni and Lachesis muta muta venoms was prolonged after incubation with either extract. The extracts also exerted antigenotoxic action on DNA fragmentation induced by L. muta muta venom. Both extracts lowered the enzymatic activity of α-glucosidases, and the ethanolic extract inhibited α-amylases. The inhibition of digestive enzymes suggests the extracts could decrease calorie availability from carbohydrate intake. Results confirm the potential therapeutic use of E. macrophyllus in the prevention and treatment of hemostatic changes, since the proteins of venoms inhibited by it exhibit high homology with human proteins involved in various pathophysiological processes, such as inflammatory responses and blood coagulation.

In vitro Assessment of Antioxidant, Membrane Stabilization and Thrombolytic Activities of Ethanolic Extract of Lagenaria siceraria Fruit

Lagenaria siceraria is a well-known vegetable fruit in Bangladesh. This study aimed to investigate the antioxidant, membrane stabilizing and thrombolytic activities of the ethanolic extract of L. siceraria fruits (EELSF). Total phenolic content and DPPH free radical scavenging activity tests were done to determine antioxidant properties. The anti-inflammatory effects were evaluated using heat- and hypotonic solution induced hemolysis. Human RBC from healthy volunteers were used to assess the thrombolytic activity of the extract. Ethanolic extract of L. siceraria fruits displayed noticeable antioxidant, membrane stabilizing and thrombolytic activities when compared to the standard drugs. Further studies are necessary to evaluate the pharmacological activities of different solvent fractions and plant parts of L. siceraria. Bangladesh Pharmaceutical Journal 27(1): 1-8, 2024 (January)

Antidepressant, Antidiarrheal, Thrombolytic and Phytochemical Profiling of Ethanol Extract of Amomum aromaticum Leaves: In vivo and In vitro Approaches

The study evaluated several pharmacological effects including antidepressant, antidiarrheal and thrombolytic activities of ethanol extract of Amomum aromaticum leaves (EEAA). Upon the assessment for qualitative phytochemical groups in the extract, several doses of EEAA (1000 - 4000 mg/kg) were studied to find acute oral toxicity in mouse for safe dose selection. Then EEAA was tested whether it demonstrates antidepressant activity in tail suspension test (TST) and forced swim test (FST). Antidiarrheal and clot lysis activities of EEAA were evaluated in castor oil-induced diarrhoea in mouse and In vitro clot lysis method, respectively. Oral administration of EEAA (1000 - 4000 mg/kg) showed no morality after 10 days, and no sign of acute toxicity observed within 24 hrs post-treatment. The qualitative phytochemical screening showed the presence of carbohydrate, alkaloid, flavonoid, tannin, saponin, and polyphenol groups in EEAA. The TST and FST resulted with significant improvement in mobility in mice treated with EEAA (400 mg/kg), where fluoxetine (20 mg/kg) was used as standard in both tests. EEAA treatment also showed a moderate dose-dependent anti-diarrheal effect. The 400 mg/kg oral dosing for 14 days decreased the rate of defecation by 52.8% compared to the control group. This study also demonstrated that EEAA possesses clot lysis activity. Hence, further intense investigations are suggested to identify specific potential active phytochemicals. Bangladesh Pharmaceutical Journal 27(1): 51-58, 2024 (January)

Anti-Arthritic, Anti-Inflammatory, Thrombolytic, Membrane Stabilizing, Antifungal and Cytotoxic Activity of Polyscias scutellaria Leaf Extract: An In-vitro Analysis

The objective of this investigations was to evaluate in Vitro anti-arthritic, anti-inflammatory, thrombolytic, membrane stabilizing, antifungal and cytotoxic activities from the methanolic leaf extract of Polyscias scutellaria. Primary evaluation of methanolic extract of Polyscias scutellaria leaf (MEPSL) was performed via phytochemical screening. Presence of Alkaloids, flavonoids, saponins, glycosides, carbohydrates and reducing sugars were observed by phytochemical screening, among other secondary compounds. Using in vitro methods of protein denaturation, the anti-arthritic and anti-inflammatory properties of MEPSL was investigated. The results showed that the extracts significantly slowed down arthritis and inflammation with the percent inhibition of 94.59% and 86.33% at the conc. of 1000 µg/mL compared to the standard diclofenac sodium (98.19%) and acetyle salicylic acid (98.56%) at same concentration respectively. The thrombolytic activity of the extracts was additionally examined by clotlysis method, and the results showed that the ability to break up blood clots increased with the amount of extract used with the value of 97.32% which is very significant compared to the standard streptokinase which showed clotlysis of 91.304%. MEPSL was also showed action that stabilized membranes by using heat induced hemolysis method, which could be helpful in treating conditions like bleeding and swelling with the percent of protection value of 58.87% when compared to standard diclofenac sodium with the percent of protection value 73.63%. Antifungal action was also seen, which shows that it could be used to treat diseases caused by fungi with the zone of inhibition 7-36 mm varying on the type of fungi. Lastly, in vitro method was used to investigate the extracts' damaging effect on Artemia salina by using shrimp lethality assay. The results showed significant cytotoxicity with the LC50 value of 1.057 µg/mL compared to the standard vincristine sulphate (LC50 value of 0.608 µg/mL). To sum up, it is clear that the phytochemical found in this plant can be used for wide range of drug discovery field due to its potent pharmacological actions.

Antidiabetic and Thrombolytic Activities of Some Selected Medicinal Plants in High Fat Diet and Dexamethasone-Induced Type 2 Diabetic Rats

Background: Type-2 diabetes (T2D) is a hypercoagulable and hypofibrinolytic condition that predisposes to cardiovascular and thrombotic complications. We screened three medicinal plants (Albizzia chevalieri, Newbouldia laevis and Leptadenia hastate) for their antidiabetic and thrombolytic activities.
 Methods: T2D was induced with high fat diet and dexamethasone. Following induction, rats were grouped into 6 (n=8 rats); control, untreated, treated (500mg/kg body weight (BW) metformin only) or treated (300mg/kg BW leaf extracts). The rats were treated for two weeks and euthanized. About 2ml of the collected blood was used for thrombolytic activity assay while the rest was processed, and the recovered serum utilised for biochemical and hormonal assays.
 Results: Rats treated with extracts had significantly (p<0.05) lowered concentrations of serum glucose, TG, LDL-cholesterol, VLDL- cholesterol and increased HDL-cholesterol compared to untreated rats. Extracts also lowered (p<0.05) the serum insulin concentration on day 14 compared to untreated rats. Albizzia chevalieri, Newbouldia laevis and Leptadenia hastata showed 48.90%, 39.20% and 37.69% clot lysis activities respectively which was significantly higher (p<0.001) than distilled water, while streptokinase produced a substantial clot lysis of 93.70%.
 Conclusions: The leaf extracts of Albizzia chevalieri, Newbouldia laevis and Leptadenia hastata have antidiabetic and thrombolytic activities in type-2 diabetic rats and thus, can be potentially used as combined antidiabetic and thrombolytic agents with in vivo effects in type-2 diabetic patients.

Integrating Phenotypic and Chemoproteomic Approaches to Identify Covalent Targets of Dietary Electrophiles in Platelets.

A large variety of dietary phytochemicals has been shown to improve thrombosis and stroke outcomes in preclinical studies. Many of these compounds feature electrophilic functionalities that potentially undergo covalent addition to the sulfhydryl side chain of cysteine residues within proteins. However, the impact of such covalent modifications on the platelet activity and function remains unclear. This study explores the irreversible engagement of 23 electrophilic phytochemicals with platelets, unveiling the unique antiplatelet selectivity of sulforaphane (SFN). SFN impairs platelet responses to adenosine diphosphate (ADP) and a thromboxane A2 receptor agonist while not affecting thrombin and collagen-related peptide activation. It also substantially reduces platelet thrombus formation under arterial flow conditions. Using an alkyne-integrated probe, protein disulfide isomerase A6 (PDIA6) was identified as a rapid kinetic responder to SFN. Mechanistic profiling studies revealed SFN's nuanced modulation of PDIA6 activity and substrate specificity. In an electrolytic injury model of thrombosis, SFN enhanced the thrombolytic activity of recombinant tissue plasminogen activator (rtPA) without increasing blood loss. Our results serve as a catalyst for further investigations into the preventive and therapeutic mechanisms of dietary antiplatelets, aiming to enhance the clot-busting power of rtPA, currently the only approved therapeutic for stroke recanalization that has significant limitations.

Evaluation of Phytochemical Screening, Antioxidant, and Thrombolytic Activity of Methanolic Extract of Phlogacanthus thyrsiflorus

Objective: The main objectives of the study are preliminary phytochemical screening, antioxidant activity and in vitro thrombolytic activity evaluation of extract Phlogacanthus thyrsiflorus. Methods: Preliminary phytochemical analysis was done by using standard methods. Human blood clot lysis was used to evaluate the thrombolytic activities. The antioxidant activity is evaluated by free radical scavenging activity of DPPH (1,1-diphenyl-2-picrylhydrazyl) method and total phenolic content. Results: Preliminary phytochemical screening revealed the presence of various bioactive components like Tannin, Saponins, Phenol and Flavonoids. In thrombolytic assay, a significant clot lysis was observed plant extract compared to the negative control. The plant could be beneficial in synthetic drug formulation by virtue of the presence of antioxidant activity. The crude extract of plant were found to have Antioxidant activity with the concentration of 20 µg/ml, 40 µg/ml, 60 µg/ml, 80 µg/ml, 100 µg/ml respectively compared to control and standard. In DPPH scavenging assay, IC50 values of extract and ascorbic acid were found to be 31.87 µg/ml and 52.04 µg/ml respectively. Total phenol content was 10.5 mg. Conclusion: In conclusion, with this study it could be concluded that the phytochemical, antioxidant assay and thrombolytic activity of methanol extract Phlogacanthus thyrsiflorus would give valuable information for further characterization and exploitation of this important medicinal plant.

Nanomicrosphere sustained-release urokinase systems with antioxidant properties for deep vein thrombosis therapy.

Deep vein thrombosis (DVT) is a venous return disorder caused by abnormal clotting of blood in deep veins. After thrombosis, most of the thrombus will spread to the deep vein trunk throughout the limb. If DVT is not treated in time, most of them will develop into thrombosis sequelae and even threaten life. Intravenous thrombolytic drugs are the most promising strategy for treating DVT, but current drugs used for thrombolysis suffer from short half-lives and narrow therapeutic indexes. To effectively manage DVT, it is necessary to develop a novel multifunctional drug-loading system to effectively prolong the treatment time and improve the therapeutic efficacy. In this study, a urokinase-loaded protocatechuic aldehyde-modified chitosan microsphere drug-loading platform was constructed for the treatment of DVT. This microsphere adsorbed urokinase well through electrostatic interaction, and the introduction of bovine serum albumin conferred stability to the microspheres. Therefore, the microsphere drug delivery system could achieve slow drug release to effectively dissolve blood fibrin. In addition, chitosan grafted with protocatechuic aldehyde imparted excellent antioxidant activity to the system to reduce free radicals in the blood vessels. Effective management of oxidative stress could avoid abnormal platelet activation and new thrombus formation. The experimental results showed that this microsphere had good biocompatibility, anti-inflammatory properties, and considerable thrombolytic activity. In conclusion, this study provided a new direction and developed a novel multi-functional nano microsphere drug delivery platform for the treatment of DVT.

Deciphering In Vitro and In Vivo Pharmacological Properties of Seed and Fruit Extracts of Flacourtia jangomas (Lour.) Raeusch.

The objective of the study was to evaluate the pharmacological properties of the methanolic extract of Flacourtia jangomas (Lour.) Raeusch fruits (PFJM) and seeds (SFJM), along with their soluble fractions in ethyl acetate (fruit: PFJE; seed: SFJE) and chloroform (fruit: PFJC; seed: SFJC). Our phytochemical analysis of the examined extracts confirmed the presence of various therapeutically active phytoconstituents, including flavonoids, tannins, glycosides, and alkaloids. Employing the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical quenching method, SFJC exhibited the highest antioxidative potential, with an IC50 of 48.84, compared to ascorbic acid (IC50 21.77). The thrombolytic activity was assessed through rapid clot analysis of human blood samples, revealing that SFJC demonstrated the highest thrombolytic activity (60.99 ± 2.28%) compared to streptokinase (72.89 ± 2.19%). In the protein denaturation antiarthritic test, the PFJE and SFJC extracts exhibited significant potency, achieving results of 74.28 ± 1.16% and 79.25 ± 0.83%, respectively, at a dose of 500 μg/mL. All samples displayed notable anthelmintic activity by reducing Pheretima posthuma paralysis and death time in a dose-dependent manner compared to albendazole. In both in vivo analgesic tests, SFJC demonstrated substantial (p < 0.01) pain inhibition percentages (tail immersion: 49.46%; acetic acid writhing: 66.43%) at a dose of 600 mg/kg. During neuropharmacological screening, all extracts significantly (p < 0.01; p < 0.05) and dose-dependently decreased the mice's locomotion activity and motor balance. In the thiopental-induced sedation assay, SFJC significantly decreased the sleep latency time (4.18 ± 0.24 min) and increased the duration of sleep time (85.20 ± 2.39 min) at a higher dose. All samples notably reduced blood glucose levels in the oral glucose tolerance test in a dose-responsive manner, and SFJC exhibited a considerable hypoglycemic impact (7.38 ± 0.44 mmoles/L at 600 mg/kg). The frequency of diarrheal episodes in mice during the antidiarrhea assessment was significantly decreased by the tested plant samples. These findings can serve as a reference for future endeavors to isolate pure bioactive compounds from this plant for the development of novel phytomedicines.

Investigation of the pharmacological characteristics of Sphaeranthus indicus flowers using phytochemical analysis

Although the conventional usage of Sphaeranthus indicus flowers in medicine exists, there is a lack of research on the precise pharmacological characteristics and comprehensive phytochemical composition of its flowers, resulting in a knowledge gap on their whole therapeutic capabilities. The aim of this work is to identify and describe the phytochemicals found in the flowers of Sphaeranthus indicus and assess specific pharmacological effects, such as antioxidant, antibacterial, anti-inflammatory, thrombolytic, antidepressant, and antidiarrheal characteristics. The investigation revealed that the MESIF (methanol extract of S. indicus flowers) showed substantial antioxidant performance (IC50 = 26.33) accompanied by a moderate phenolic content (93.33 ± 1.76). At 250 and 500 µg/ml, the extract exhibited modest to moderate antibacterial sensitivity against many tested pathogens and significantly reduced inflammation by protein denaturation (41.63 ± 0.11 and 70.52 ± 0.20). Furthermore, MESIF exhibited remarkable thrombolytic activity with a statistically significant (***P&lt;0.001) lysis of clots. The extract showed substantial reduction in immobility in both the Forced Swim Test (FST) and Tail Suspension Test (TST) at doses of 200 mg/kg and 400 mg/kg (***p&lt;0.001). It also a showed significant potent antidiarrheal activity (**p&lt;0.01 at 200 mg/kg and ***p&lt;0.001 at 400 mg/kg), effectively reducing diarrheal episodes in the test models. These findings suggest that the MESIF holds potential for developing treatments with antioxidant, antibacterial, anti-inflammatory, thrombolytic, antidepressant, and antidiarrheal effects. Further research is recommended to isolate specific bioactive compounds and elucidate the mechanisms underlying these pharmacological actions.