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1.
J Agric Food Chem ; 68(10): 3132-3139, 2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32064873

RESUMO

Thrombin is currently one of the important targets for the treatment and prevention of thrombosis. At present, there are few reports on the application of lactoferrin peptides in anticoagulation. In this study, a peptide with the amino acid sequence of LRPVAAEIY (LF-LR) derived from lactoferrin was shown to possess antithrombotic activity. LF-LR (5 mM) significantly prolonged activated partial thromboplastin time, prothrombin time, and thrombin time for 13.4, 1.7, and 5.1 s, respectively. It prolonged the coagulation time of fibrinogen from 15.3 ± 0.4 to 20.2 ± 0.5 s by affecting the conformation of thrombin. Using circular dichroism analysis, LF-LR can increase the α-helix content of thrombin from 25.6 to 56.7% and made the ß-sheet disappear. In addition, LF-LR also quenched fluorescence of thrombin at about 346 nm (λEx = 280 nm). By means of molecular docking, it was found that LF-LR could bind to both the active site and the exosite-I of thrombin, and the combined LYS60F, TRP60D, ASP189, LYS36, and ARG77A are typical amino acids in the two domains, respectively.


Assuntos
Anticoagulantes/química , Lactoferrina/química , Peptídeos/química , Trombina/química , Sequência de Aminoácidos , Animais , Sítios de Ligação , Domínio Catalítico , Bovinos , Fibrinogênio/química , Humanos , Cinética , Simulação de Acoplamento Molecular , Ligação Proteica , Tempo de Trombina
2.
Carbohydr Polym ; 227: 115312, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31590876

RESUMO

Low molecular weight heparin (LMWH) possesses a dual function of anticoagulation and anti-inflammation. While the structures and mechanisms on its anticoagulation have been widely studied, the structural features responsible for the anti-inflammatory activity of LMWH remain to be explored. In the present study, guided by an anti-inflammation assay, a non-anticoagulant species was generated from partial desulfation of LMWH to fully retain the anti-inflammatory activity, from which five fractions were further separated and three of them were characterized by enzymatic degradation, hydrophobic labeling, C18-based HPLC and LC-MS/MS analyses. The structure-activity relationship revealed that the sulfate groups in LMWH are critical to distinguish and separate the activities of anticoagulation and anti-inflammation, leading to the identification of a synthetic heparosan-type heptasaccharide as a potent anti-inflammatory agent. The present strategy enables the simplification of complex polysaccharides to bioactive synthetic oligosaccharides for therapeutic utility.


Assuntos
Anti-Inflamatórios/farmacologia , Anticoagulantes/química , Dissacarídeos/farmacologia , Heparina de Baixo Peso Molecular/química , Sulfatos/química , Animais , Anti-Inflamatórios/química , Dissacarídeos/química , Fator Xa/química , Heparina Liase/química , Lipopolissacarídeos/farmacologia , Camundongos , Óxido Nítrico/metabolismo , Protrombina/química , Células RAW 264.7
3.
J Agric Food Chem ; 68(1): 176-184, 2020 Jan 08.
Artigo em Inglês | MEDLINE | ID: mdl-31850760

RESUMO

Thrombin can be used as a target for its inhibitors to prevent blood coagulation. A novel peptide (TKLTEEEKNR, PfCN) identified from αS2-casein (fragments 211-220) with high anticoagulant activity was screened and prepared. The activated partial thromboplastin time, prothrombin time, and thrombin time, at the concentration of 4 mM, prolonged about 19, 2.5 and 5.5 s, respectively. At the same concentration, the fibrinogen clotting time prolonged from 25.5 ± 0.7 to 38.3 ± 1.3 s. The thrombin inhibitory efficiency in vitro (IC50 value of 29.27 mM) and antithrombosis effect in vivo were determined. The secondary structure of thrombin, which was influenced by PfCN, indicates that PfCN can bind to thrombin. Isothermal titration calorimetry and the chromogenic substrate test showed that PfCN belongs to the bivalent thrombin inhibitor like bivalirudin. Although the effect was not as good as bivalirudin, in the animal experiment, bleeding occurred in the bivalirudin group but not in the PfCN group. Moreover, molecular docking illustrates the mechanism for the antithrombin activity of PfCN. These results indicated that PfCN could be used as an effective thrombin inhibitor with broad potential for the prevention of thrombotic acute pulmonary embolism and other thrombotic events.


Assuntos
Anticoagulantes/química , Peptídeos/química , Trombina/química , Animais , Anticoagulantes/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Domínio Catalítico , Bovinos , Humanos , Cinética , Simulação de Acoplamento Molecular , Tempo de Tromboplastina Parcial , Peptídeos/farmacologia , Tempo de Protrombina
4.
Biochem Med (Zagreb) ; 30(1): 010702, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31839722

RESUMO

Introduction: Clinical application of rivaroxaban and apixaban does not require therapeutic monitoring. Commercial anti-activated factor X (anti-FXa) inhibition methods for all anti-FXa drugs are based on the same principle, so there are attempts to evaluate potential clinical application of heparin-calibrated anti-FXa assay as an alternative method for direct FXa inhibitors. We aimed to evaluate relationship between anti-FXa methods calibrated with low molecular weight heparin (LMWH) and with drug specific calibrators, and to determine whether commercial LMWH anti-FXa assay can be used to exclude the presence of clinically relevant concentrations of rivaroxaban and apixaban. Materials and methods: Low molecular weight heparin calibrated reagent (Siemens Healthineers, Marburg, Germany) was used for anti-FXa activity measurement. Innovance heparin (Siemens Healthineers, Marburg, Germany) calibrated with rivaroxaban and apixaban calibrators (Hyphen BioMed, Neuville-sur-Oise, France) was used for quantitative determination of FXa inhibitors. Results: Analysis showed good agreement between LMWH calibrated and rivaroxaban calibrated activity (κ = 0.76) and very good agreement with apixaban calibrated anti-Xa activity (κ = 0.82), respectively. Low molecular weight heparin anti-FXa activity cut-off values of 0.05 IU/mL and 0.1 IU/mL are suitable for excluding the presence of clinically relevant concentrations (< 30 ng/mL) of rivaroxaban and apixaban, respectively. Concentrations above 300 ng/mL exceeded upper measurement range for LMWH anti-FXa assay and cannot be determined by this method. Conclusion: Low molecular weight heparin anti-FXa assay can be used in emergency clinical conditions for ruling out the presence of clinically relevant concentrations of rivaroxaban and apixaban. However, use of LMWH anti-FXa assay is not appropriate for their quantitative determination as an interchangeable method.


Assuntos
Anticoagulantes/química , Testes de Coagulação Sanguínea/métodos , Heparina de Baixo Peso Molecular/química , Pirazóis/química , Piridonas/química , Rivaroxabana/química , Anticoagulantes/metabolismo , Área Sob a Curva , Testes de Coagulação Sanguínea/normas , Calibragem , Compostos Cromogênicos/química , Fator Xa/química , Fator Xa/metabolismo , Inibidores do Fator Xa/química , Inibidores do Fator Xa/metabolismo , Heparina de Baixo Peso Molecular/metabolismo , Humanos , Pirazóis/metabolismo , Piridonas/metabolismo , Curva ROC
5.
Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi ; 37(11): 852-854, 2019 Nov 20.
Artigo em Chinês | MEDLINE | ID: mdl-31826554

RESUMO

Early hemoperfusion in poisoned patients can remove poisons rapidly and effectively, which plays an important role in improving the prognosis of patients. The key of hemoperfusion therapy is the safe and effective anticoagulation. The local citrate anticoagulation effect acid is good, it also has little effect on the systemic coagulation mechanism and internal environment of patients, so it is worthy of promotion. We retrospectively analyzed the clinical data and treatment of 273 patients who were poisoned by citrate anticoagulant in the emergency intensive care unit of the Second Affiliated Hospital of Shandong First Medical University, aiming at perfusion of citrate anticoagulant in patients with poisoning. Provide a certain clinical reference.


Assuntos
Anticoagulantes , Ácido Cítrico , Hemoperfusão , Envenenamento , Anticoagulantes/administração & dosagem , Anticoagulantes/química , Ácido Cítrico/administração & dosagem , Ácido Cítrico/química , Hemoperfusão/normas , Humanos , Envenenamento/terapia , Venenos/química , Estudos Retrospectivos
6.
Int J Mol Sci ; 20(19)2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31574907

RESUMO

Snake venom enzymes of the L-amino acid oxidase (LAAO) class are responsible for tissue hemorrhage, edema, and derangement of platelet function. However, what role, if any, these flavoenzymes play in altering plasmatic coagulation have not been well defined. Using coagulation kinetomic analyses (thrombelastograph-based), it was determined that the LAAO derived from Crotalus adamanteus venom displayed a procoagulant activity associated with weak clot strength (no factor XIII activation) similar to thrombin-like enzymes. The procoagulant activity was not modified in the presence of reduced glutathione, demonstrating that the procoagulant activity was likely due to deamination, and not hydrogen peroxide generation by the LAAO. Further, unlike the raw venom of the same species, the purified LAAO was not inhibited by carbon monoxide releasing molecule-2 (CORM-2). Lastly, exposure of the enzyme to phenylmethylsulfonyl fluoride (PMSF) resulted in the LAAO expressing anticoagulant activity, preventing contact activation generated thrombin from forming a clot. In sum, this investigation for the first time characterized the LAAO of a snake venom as both a fibrinogen polymerizing and an anticoagulant enzyme acting via oxidative deamination and not proteolysis as is the case with thrombin-like enzymes (e.g., serine proteases). Using this thrombelastographic approach, future investigation of purified enzymes can define their biochemical nature.


Assuntos
Crotalus , L-Aminoácido Oxidase/metabolismo , L-Aminoácido Oxidase/farmacologia , Venenos de Serpentes/enzimologia , Animais , Anticoagulantes/química , Anticoagulantes/metabolismo , Anticoagulantes/farmacologia , Coagulação Sanguínea/efeitos dos fármacos , Cálcio/metabolismo , Cálcio/farmacologia , Coagulantes/química , Coagulantes/metabolismo , Coagulantes/farmacologia , Ácido Edético/farmacologia , Glutationa/metabolismo , Glutationa/farmacologia , Heparina/farmacologia , Humanos , Cinética , L-Aminoácido Oxidase/química , Compostos Organometálicos/farmacologia , Tromboelastografia
7.
Pak J Pharm Sci ; 32(4): 1581-1588, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31608877

RESUMO

Warfarin is administered as a racemic preparation of R- and S-enantiomers. S-warfarin is more potent than R-warfarin, so changes in blood levels of S-warfarin affect the anticoagulant response. This study was carried out to determine the effect of CYP2C9*2 and CYP2C9*3 polymorphisms on S/R warfarin ratio. A single blood sample was collected 12-16 hours after drug administration from 170 stable patients fulfilling the criteria. Genotyping of the CYP2C9 polymorphisms was done by polymerase chain reaction-restriction fragment length polymorphism assay. S- and R-warfarin enantiomers extraction from plasma was accomplished by a validated HPLC method. The concentration of S-warfarin was significantly different among CYP2C9 genotypes (p =0.018) whereas there was no effect on R-warfarin (p =0.134). There was statistically significant effect of different CYP2C9 genotypes on S/R warfarin ratio (p=0.000). It is concluded that CYP2C9 polymorphisms influence CYP2C9 enzymatic activity in turn affecting S-warfari levels but not R-warfarin, thus leading to different S/R warfarin enantiomers ratio among different CYP2C9 genotypes.


Assuntos
Citocromo P-450 CYP2C9/genética , Varfarina/química , Varfarina/farmacocinética , Adolescente , Adulto , Idoso , Anticoagulantes/administração & dosagem , Anticoagulantes/sangue , Anticoagulantes/química , Anticoagulantes/farmacocinética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Paquistão , Polimorfismo Genético , Polimorfismo de Fragmento de Restrição , Estereoisomerismo , Varfarina/administração & dosagem , Varfarina/sangue , Adulto Jovem
8.
J Oleo Sci ; 68(10): 959-965, 2019 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-31511465

RESUMO

Protein hydrolysis on the freeze-thaw stability of emulsions prepared with soy protein - dextran conjugates were investigated. Soy protein isolate-dextran (SPI-D) and soy protein hydrolysates-dextran (SPH-D) conjugates with different degree of hydrolysis (DH) were formed by Maillard reaction. The formation of protein-polysaccharide conjugates between SPI/SPH and dextran molecules was confirmed by SDS-PAGE; this finding was consistent with the degree of glycation and the browning index. The freeze-thaw emulsion stability was investigated. The results confirmed that the SPH3-D (DH at 3%) emulsion with 3% DH of SPI exhibited the lowest creaming index after experiencing 1, 2, and 3 freeze-thaw cycles , with results of 7.69%, 20.74% and 31.30%, respectively. The SPH3-D emulsion had a significantly lower average particle size, which was reduced by 48.28% compared to the SPI-D emulsion. Meanwhile, the SPH3-D solution had low interfacial tension. The confocal laser scanning microscopy analysis indicated that the SPH3-D emulsions were strongly stable against the freeze-thaw treatment and could be used as effective emulsifiers in frozen foods.


Assuntos
Anticoagulantes/química , Dextranos/química , Congelamento , Proteínas de Soja/química , Emulsões , Hidrólise , Estabilidade Proteica , Proteínas de Soja/isolamento & purificação
9.
Eur J Pharm Sci ; 139: 105066, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31513922

RESUMO

Thrombomodulin (TM) is an endothelial cell membrane protein that plays essential roles in controlling vascular haemostatic balance. The 4, 5, 6 EGF-like domain of TM (TM456) has cofactor activity for thrombin binding and subsequently protein C activation. Therefore, recombinant TM456 is a promising anticoagulant candidate but has a very short half-life. Ligation of poly (ethylene glycol) to a bioactive protein (PEGylation) is a practical choice to improve stability, extend circulating life, and reduce immunogenicity of the protein. Site-specific PEGylation is preferred as it could avoid the loss of protein activity resulting from nonspecific modification. We report herein two site-specific PEGylation strategies, enzymatic ligation and copper-free click chemistry (CFCC), for rTM456 modification. Recombinant TM456 with a C-terminal LPETG tag (rTM456-LPETG) was expressed in Escherichia coli for its end-point modification with NH2-diglycine-PEG5000-OMe via Sortase A-mediated ligation (SML). Similarly, an azide functionality was easily introduced at the C-terminus of rTM456-LPETG via SML with NH2-diglycine-PEG3-azide, which facilitates a site-specific PEGylation of rTM456via CFCC. Both PEGylated rTM456 conjugates retained protein C activation activity as that of rTM456. Also, they were more stable than rTM456 in Trypsin digestion assay. Further, both PEGylated rTM456 conjugates showed a concentration-dependent prolongation of thrombin clotting time (TCT) compared to non-modified protein, which confirms the effectiveness of these two site-specific PEGylation schemes.


Assuntos
Anticoagulantes/administração & dosagem , Anticoagulantes/química , Trombomodulina/administração & dosagem , Trombomodulina/química , Azidas/administração & dosagem , Azidas/química , Coagulação Sanguínea/efeitos dos fármacos , Química Click , Estabilidade de Medicamentos , Humanos , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/química , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/química , Trombina/metabolismo , Trombomodulina/genética
10.
BMC Complement Altern Med ; 19(1): 236, 2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31481052

RESUMO

BACKGROUND: Syzygium cumini (L.) Skeels. is one of the very popular traditionally used medicinal plants with numerous pharmacological activities including antioxidant, hypoglycemic and anti-inflammatory. However, actions of S. cumini on blood coagulation and other parameters of blood were poorly pharmacologically studied. Therefore, aim of this present investigation is to examine the effects of methanolic extract of S. cumini on blood coagulation and anticoagulation factors in healthy white albino rabbits at different doses. METHODS: Blood samples were drawn twice during this study and biochemical assays were performed to determine the effect on different parameters such as coagulation, anticoagulation, hematological, Protein C (PC) and thrombin antithrombin (TAT) complex and platelet aggregation. RESULTS: The results showed significant increase in RBCs, hemoglobin, hematocrit and platelets counts up to 1.4 × 103/cm, 2.2 g/dl, 6%, 248.2 × 103/cm respectively. While, thrombin and bleeding time were also prolonged in dose dependent manner which is highly significant (p ≤ 0.005) as compared to control. Similarly, highly significantly increased (p ≤ 0.005) in levels of protein C, thrombin antithrombin complex at dose of 500 mg/kg were observed. Whereas, levels of platelets aggregation and fibrinogen were decreased at high doses. CONCLUSION: The obtained findings of hematological and coagulation tests concludes possibly S. cumini possess anticoagulant and antiplatelet effects.


Assuntos
Anticoagulantes/farmacologia , Extratos Vegetais/farmacologia , Inibidores da Agregação de Plaquetas/farmacologia , Syzygium/química , Animais , Anticoagulantes/química , Coagulação Sanguínea/efeitos dos fármacos , Masculino , Extratos Vegetais/química , Folhas de Planta/química , Agregação Plaquetária/efeitos dos fármacos , Inibidores da Agregação de Plaquetas/química , Coelhos
11.
Arch Pharm Res ; 42(10): 862-878, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31493264

RESUMO

Ginseng is the most frequently used herbal medicine for immune system stimulation and as an adjuvant with prescribed drugs owing to its numerous pharmacologic activities. It is important to investigate the beneficial effects and interaction of ginseng with therapeutic drugs. This review comprehensively discusses drug metabolizing enzyme- and transporter-mediated ginseng-drug interaction by analyzing in vitro and clinical results with a focus on ginsenoside, a pharmacologically active marker of ginseng. Impact of ginseng therapy or ginseng combination therapy on diabetic patients and of ginseng interaction with antiplatelets and anticoagulants were evaluated based on ginseng origin and ginsenoside content. Daily administration of Korean red ginseng (0.5-3 g extract; dried ginseng > 60%) did not cause significant herb-drug interaction with drug metabolizing enzymes and transporters. Among various therapeutic drugs administered in combination with ginseng, adjuvant chemotherapy, comprising ginseng (1-3 g extract) and anticancer drugs, was effective for reducing cancer-related fatigue and improving the quality of life and emotional scores. Limited information regarding ginsenoside content in each ginseng product and plasma ginsenoside concentration among patients necessitates standardization of ginseng product and establishment of pharmacokinetic-pharmacodynamic correlation to further understand beneficial effects of ginseng-therapeutic drug interactions in future clinical studies.


Assuntos
Anticoagulantes/farmacologia , Antineoplásicos Fitogênicos/farmacologia , Ginsenosídeos/farmacologia , Hipoglicemiantes/farmacologia , Neoplasias/tratamento farmacológico , Panax/química , Inibidores da Agregação de Plaquetas/farmacologia , Anticoagulantes/química , Antineoplásicos Fitogênicos/química , Diabetes Mellitus/tratamento farmacológico , Ginsenosídeos/química , Interações Ervas-Drogas , Humanos , Hipoglicemiantes/química , Inibidores da Agregação de Plaquetas/química
12.
Carbohydr Polym ; 224: 115146, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31472868

RESUMO

Fucosylated glycosaminoglycan (FG), a glycosaminoglycan derivative containing distinct sulfated fucose (FucS) branches, displays potent anticoagulant activity by inhibiting the intrinsic tenase complex (iXase). Herein, AmFG, SvFG and HaFG from three species of sea cucumbers were isolated and depolymerized by ß-eliminative cleavage. Three series of fragments, A1-A4, S1-S4 and H1-H4, were purified from the depolymerized FGs. Based on structural analysis of these fragments, three FGs were deduced as -{→4)-[L-FucS-α(1→3)]-D-GlcA-ß(1→3)-D-GalNAc4S6S-ß(1}n-. The structures differed in sulfation types of FucS, namely, most of FucS in AmFG was Fuc3S4S, but the FucS in SvFG was Fuc2S4S, while the FucS in HaFG was Fuc3S4S, Fuc2S4S and Fuc4S. However, all FucS branches attached to C-3 of GlcA as monosaccharides. Anticoagulant and anti-iXase assays showed the octasaccharide is the minimum fragment for potent anticoagulant activity via anti-iXase irrespective of FucS types. Among FG fragments with same degree of polymerization, oligosaccharides containing Fuc2S4S had more potent anti-iXase activity.


Assuntos
Inibidores de Cisteína Proteinase/química , Inibidores de Cisteína Proteinase/farmacologia , Fucose/química , Glicosaminoglicanos/química , Glicosaminoglicanos/farmacologia , Proteínas de Neoplasias/antagonistas & inibidores , Anticoagulantes/química , Anticoagulantes/farmacologia , Sequência de Carboidratos , Cisteína Endopeptidases
13.
Ann Biol Clin (Paris) ; 77(4): 391-396, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31418700

RESUMO

The SFBC working group aimed to deal with biological tests outside the French nomenclature that may be useful for the follow-up of dialysis patients. Our discussion was divided into 3 parts: 1) evaluation of peritoneal membrane characteristics; 2) monitoring of renal replacement therapy using regional citrate anticoagulation; 3) estimation of residual renal function (RRF). International recommendations underline the importance of assessing peritoneal membrane characteristics for peritoneal dialysis prescription. This peritoneal equilibrium test requires the measurement in dialysate of the following parameters: glucose, urea, creatinine and sodium. As part of the monitoring of continuous renal replacement therapy using regional citrate anticoagulation, the determination of ionized calcium assay is essential according to national and international guidelines to ensure a balance between effective anticoagulation and appropriate calcium levels. Finally, the RRF plays a key role in the dialysis adequacy and patient survival. European and international recommendations highlight the potential interest of RRF in peritoneal dialysis and hemodialysis. The RRF corresponds to the mean of urinary urea and creatinine clearance, assessed from a urine collection with measurement of urinary urea.


Assuntos
Testes de Função Renal/métodos , Rim/fisiopatologia , Monitorização Fisiológica/métodos , Peritônio/fisiologia , Diálise Renal/métodos , Anticoagulantes/química , Anticoagulantes/metabolismo , Cálcio/metabolismo , Ácido Cítrico/química , Ácido Cítrico/metabolismo , Humanos , Rim/metabolismo , Falência Renal Crônica/diagnóstico , Falência Renal Crônica/fisiopatologia , Diálise Peritoneal/métodos
14.
Molecules ; 24(16)2019 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-31426507

RESUMO

Nowadays, pharmaceutical heparin is purified from porcine and bovine intestinal mucosa. In the past decade there has been an ongoing concern about the safety of heparin, since in 2008, adverse effects associated with the presence of an oversulfated chondroitin sulfate (OSCS) were observed in preparations of pharmaceutical porcine heparin, which led to the death of patients, causing a global public health crisis. However, it has not been clarified whether OSCS has been added to the purified heparin preparation, or whether it has already been introduced during the production of the raw heparin. Using a combination of different analytical methods, we investigate both crude and final heparin products and we are able to demonstrate that the sulfated contaminants are intentionally introduced in the initial steps of heparin preparation. Furthermore, the results show that the oversulfated compounds are not structurally homogeneous. In addition, we show that these contaminants are able to bind to cells in using well known heparin binding sites. Together, the data highlights the importance of heparin quality control even at the initial stages of its production.


Assuntos
Anticoagulantes/isolamento & purificação , Sulfatos de Condroitina/isolamento & purificação , Contaminação de Medicamentos , Heparina/isolamento & purificação , Animais , Anticoagulantes/química , Bovinos , Sulfatos de Condroitina/química , Heparina/química , Heparina Liase/química , Humanos , Hidrólise , Mucosa Intestinal/química , Espectroscopia de Ressonância Magnética , Polissacarídeo-Liase/química , Controle de Qualidade , Suínos
15.
Int J Mol Sci ; 20(17)2019 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-31450643

RESUMO

The interplay between oxidative stress, inflammation, and tissue fibrosis leads to the progression of chronic kidney disease (CKD). Edoxaban, an activated blood coagulation factor Xa (FXa) inhibitor, ameliorates kidney disease by suppressing inflammation and tissue fibrosis in animal models. Interestingly, rivaroxaban, another FXa inhibitor, suppresses oxidative stress induced by FXa. Thus, FXa inhibitors could be multitargeted drugs for the three aforementioned risk factors for the progression of CKD. However, the exact mechanism responsible for eliciting the antioxidant effect of FXa inhibitors remains unclear. In this study, the antioxidant effect of edoxaban was evaluated. First, the intracellular antioxidant properties of edoxaban were evaluated using human proximal tubular cells (HK-2 cells). Next, direct radical scavenging activity was measured using the electron spin resonance and fluorescence analysis methods. Results show that edoxaban exhibited antioxidant effects on oxidative stress induced by FXa, indoxyl sulfate, and angiotensin II in HK-2 cells, as well as the FXa inhibitory activity, was involved in part of the antioxidant mechanism. Moreover, edoxaban exerted its antioxidative effect through its structure-specific direct radical scavenging activity. Edoxaban exerts antioxidant effects by inhibiting FXa and through direct radical-scavenging activity, and thus, may serve as multitargeted drugs for the three primary risk factors associated with progression of CKD.


Assuntos
Inibidores do Fator Xa/farmacologia , Depuradores de Radicais Livres/farmacologia , Piridinas/farmacologia , Tiazóis/farmacologia , Anticoagulantes/química , Anticoagulantes/farmacologia , Linhagem Celular , Espectroscopia de Ressonância de Spin Eletrônica , Inibidores do Fator Xa/química , Depuradores de Radicais Livres/química , Humanos , Radical Hidroxila/antagonistas & inibidores , Estresse Oxidativo/efeitos dos fármacos , Piridinas/química , Espécies Reativas de Oxigênio/metabolismo , Tiazóis/química
16.
Colloids Surf B Biointerfaces ; 181: 963-972, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31382347

RESUMO

There is an urgent clinical demand to develop a functional small diameter vascular graft with long-term patency, which mainly relies on the anticoagulation and endothelialization of the vascular graft. In addition, improved degradation of vascular graft provides more space for cell infiltration and facilitates remodeling of blood vessel. In this study, an elastic and biomimetic nanofibrous vascular scaffold with improved degradability was prepared by adding poly(lactic-co-glycolic acid) (PLGA) into the poly(L-lactic acid) (PLLA)/poly(L-lactide-co-ε-caprolactone) (PLCL) blend using thermally induced phase separation (TIPS) technique. The incorporation of PLGA also improved the hydrophilicity of the composite scaffold. Then the vascular scaffold was surface modified by combination of heparin and stromal cell-derived factor-1 alpha (SDF-1α). The results of whole blood clotting kinetics and plasma recalcification profiles indicated that heparinized modification significantly enhanced the anticoagulation of vascular scaffold. In vitro cell culture assays demonstrated the immobilized SDF-1α facilitated recruitment of endothelial progenitor cells (EPCs), migration and proliferation of mature endothelial cells, human umbilical vein endothelial cells (HUVECs), and expression of VE-cadherin/CD144 and endothelial nitric oxide synthase (eNOS) genes in HUVECs, thereby accelerating the endothelialization of the modified vascular scaffold. Besides, the nanofibrous vascular scaffold modified with heparin and SDF-1α inhibited the proliferation of human vascular smooth muscle cells (HVSMCs). Therefore, the phase separated nanofibrous vascular scaffold modified with heparin and SDF-1α shows the promising in vitro performance as a functional small diameter vascular graft.


Assuntos
Anticoagulantes/farmacologia , Quimiocina CXCL12/farmacologia , Heparina/farmacologia , Nanofibras/química , Polímeros/farmacologia , Anticoagulantes/química , Coagulação Sanguínea/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Quimiocina CXCL12/química , Células Endoteliais/efeitos dos fármacos , Heparina/química , Humanos , Tamanho da Partícula , Polímeros/química , Propriedades de Superfície , Engenharia Tecidual
17.
Carbohydr Polym ; 222: 115010, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31320102

RESUMO

Two sulfated polysaccharides (SPs), F2 and F3, isolated from Codium isthmocladum were found to contain galactose, sulfate, and pyruvate. The apparent molecular weights of F2 and F3 were determined to be 62 and 61 kDa, respectively. NMR spectroscopy combined with chemical analysis showed that F2 and F3 have the same structural features. However, F3 showed higher sulfate/sugar ratio (1/2.6) than F2 (1/4). F2 and F3 are essentially (1 → 3)-ß-D-galactans with some branching at C6. Pyruvylation occurs at O3 and O4, forming 3,4-O-(1-carboxyethylidene)-ß-D-Galp residues; some of these pyruvylated residues contain sulfate groups at C6. Some non-branching residues contain sulfate at C4. None of the SPs exhibited antioxidant activity. MTT results indicated that 1 mg/mL of both SPs about 40% of PANC-1 cell viability. At 10 µg/mL, F2 and F3 had 1.7-fold longer clotting times compared to that of Clexane® at the same concentration. The higher sulfate content of F3 is not a determining factor for pharmacological activities of galactans, considering that both F2 and F3 exerted the effects.


Assuntos
Anticoagulantes/farmacologia , Antioxidantes/farmacologia , Clorófitas/química , Galactanos/farmacologia , Alga Marinha/química , Anticoagulantes/química , Anticoagulantes/isolamento & purificação , Antioxidantes/química , Antioxidantes/isolamento & purificação , Sequência de Carboidratos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Galactanos/química , Galactanos/isolamento & purificação , Humanos , Piruvatos/química , Piruvatos/isolamento & purificação , Piruvatos/farmacologia , Ésteres do Ácido Sulfúrico/química , Ésteres do Ácido Sulfúrico/isolamento & purificação , Ésteres do Ácido Sulfúrico/farmacologia
18.
J Ethnopharmacol ; 242: 112046, 2019 Oct 05.
Artigo em Inglês | MEDLINE | ID: mdl-31279070

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: In Colombia, the only authorized treatment to cure snakebite envenomation is with the use of antivenom. The antivenom neutralizes the systemic effects properly, but is not very effective at neutralizing local effects, thus several cases have lead to complications. On the other hand, rural communities turn to the use of plants that are easily accessible and available for basic health care. One of these plants is named Piper auritum (PA), which is traditionally highlighted in some indigenous communities of Antioquia and Chocó. AIM OF THE STUDY: The main objective of this work was to characterize the venom's toxicity by determining the Minimum Edema Dose (MED), the Minimum Coagulant Dose-Plasma (MCD-P), the Minimum Hemorrhagic Dose (MHD) and to determine the neutralizing power of the Total Ethanolic Extract (TEE) from leaves of PA on the localized and systemic effects caused by the Bothrops rhombeatus venom. MATERIALS AND METHODS: To begin, the minimum dose that causes edema-forming, coagulant and hemorrhagic activities was determined. The protocols investigated include coagulant and edematic activities caused by the venom of Bothrops rhombeatus which were neutralized by the TEE of PA. RESULTS: The MCD-P was found to be 0.206 ±â€¯0.026 µg, the MED is the same at 0.768 ±â€¯0.065 µg, and the MHD is 3.553 ±â€¯0.292 µg, which are different from the reports for Bothrops asper and Bothrops ayerbei. Next, a phytochemical screening was done to the TEE where mainly triterpenes, steroids, coumarins, saponins, and lignans were identified. Also present were 43,733 ±â€¯2106 mg AG/g ES of phenols, which are secondary metabolites that are probably responsible for the neutralization of coagulant and edematic activities at rates of 2363.870 µL and 1787.708 µL of extract/mg of venom, respectively. As a comparative parameter, the National Institute Health's (NHI) effective dose of the antivenom was used as a comparative parameter. In addition, we determined the toxicity of the TEE of PA on to Artemia salina, being moderately toxic at 6 and 24 h, while the essential oil of PA at the same observation hours is in the extremely toxic range. CONCLUSIONS: The results reflect that the extract of P. auritum has an anti-inflammatory effect similar to that of the NIH serum. It could be used as a complement of NIH antivenom, using them together so it contributes to effectively reduce inflammation and the socio-economic impact generated by the permanence of a patient victim of snakebite in health centers. CLASSIFICATIONS: Immunological products and vaccines.


Assuntos
Anti-Inflamatórios/uso terapêutico , Anticoagulantes/uso terapêutico , Antivenenos/uso terapêutico , Venenos de Crotalídeos/toxicidade , Piper , Extratos Vegetais/uso terapêutico , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/farmacologia , Anticoagulantes/química , Anticoagulantes/farmacologia , Antivenenos/química , Antivenenos/farmacologia , Artemia/efeitos dos fármacos , Coagulação Sanguínea/efeitos dos fármacos , Bothrops , Edema/tratamento farmacológico , Etanol/química , Hemorragia/tratamento farmacológico , Camundongos Endogâmicos ICR , Compostos Fitoquímicos/análise , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/uso terapêutico , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Mordeduras de Serpentes/tratamento farmacológico , Solventes/química
19.
J Pharm Biomed Anal ; 174: 639-643, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31279893

RESUMO

Heparin is a carbohydrate polymer, which is clinically used as an anticoagulant for the treatment of thrombotic disorders. The anticoagulant process is mainly mediated by the interaction of heparin with antithrombin followed by inhibition of clotting factors IIa (FIIa) and Xa (FXa). The influence of polymer disaccharide structure, average molecular weight and impurity profiling (e.g., chloride and water content) was investigated by NMR spectrometry and principal component analysis (PCA) for a representative dataset of porcine heparin samples (n = 509). A significant linear dependence was found between anticoagulant activity and scores on the third principal component (PC3) based on the non-targeted analysis of 1H NMR fingerprints. The correlation between average molecular values and anticoagulant activity for the 24 porcine heparin samples from two manufacturers was linear (R = 0.85) over typical values for porcine heparin preparations. Chloride and water contents were identified as negatively influencing factors for the actual activity values as their presence decrease the "pharmaceutically active" organic part of heparin preparations. Some suggestions regarding manufacturing process are made according to the results.


Assuntos
Anticoagulantes/análise , Heparina/análise , Espectroscopia de Ressonância Magnética , Animais , Anticoagulantes/química , Cloretos/química , Dissacarídeos/química , Fator Xa/química , Inibidores do Fator Xa/análise , Inibidores do Fator Xa/química , Heparina/química , Heparina de Baixo Peso Molecular/análise , Modelos Lineares , Peso Molecular , Polímeros/química , Análise de Componente Principal , Protrombina/química , Suínos , Água/química
20.
Mater Sci Eng C Mater Biol Appl ; 103: 109769, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349444

RESUMO

Non-covalent electrostatic interaction between amide nitrogen and carbonyl carbon of shorter chain length of polyvinylpyrrolidone (PVP-k25) was developed with in-house carboxylic oxidized multiwall carbon nanotubes (O-MWCNT) and then blended with Polyethersulfone (PES) polymer. FTIR analysis was utilized to confirm bonding nature of nano-composites (NCs) of O-MWCNT/PVP-k25 and casting membranes. Non-solvent induces phase separation process developed regular finger-like channels in composite membranes whereas pristine PES exhibited spongy entities as studied by cross sectional analysis report of FESEM. Further, FESEM instrument was also utilized to observe the dispersion of O-MWCNT/PVP based nanocomposite (NCs) with PES and membranes leaching phenomena analysis. Contact angle experiments described 24% improvement of hydrophilic behaviour, leaching ratio of additives was reduced to 1.89%, whereas water flux enhanced up to 6 times. Bovine serum albumin (BSA) and lysozyme based antifouling analysis shown up to 25% improvement, whereas 84% of water flux was regained after protein fouling than pristine PES. Anticoagulant activity was reported by estimating prothrombin, thrombin, plasma re-calcification times and production of fibrinogen cluster with platelets-adhesions photographs and hemolysis experiments. Composite membranes exhibited 3.4 and 3 times better dialysis clearance ratios of urea and creatinine solutes as compared to the raw PES membrane.


Assuntos
Anticoagulantes/química , Membranas Artificiais , Nanocompostos/química , Nanotubos/química , Polímeros/química , Diálise Renal/instrumentação , Sulfonas/química , Humanos
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