The recombinant plant Bauhinia bauhinioides elastase inhibitor reduces rat thrombus without alterations in hemostatic parameters

Cleide Oliveira,Misako Uemura Sampaio,Maria Luiza Vilela Oliva,Francisco Humberto De Abreu Maffei,Tatiana Fontes Ottaiano,Antonio Miranda,Mayara Vioto Valois,Daiane Hansen

doi:10.1038/s41598-021-92745-4

Cleide Oliveira, Misako Uemura Sampaio + Show 6 more

Open Access

https://doi.org/10.1038/s41598-021-92745-4

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Abstract

The anti-inflammatory effects of the plant protease inhibitor BbCI (Bauhinia bauhinioides cruzipain inhibitor), which blocks elastase, cathepsin G, and L, and proteinase 3 has been demonstrated. Here, we investigated the recombinant rBbCI-His(6) (containing a histidine tail) in an experimental venous thrombosis model of vena cava (VC) ligature in rats, comparing to heparin. We evaluate the effects of the inhibitors (native or recombinant) or heparin on the activated partial thromboplastin time (aPTT) and prothrombin time (PT) in human and rat plasmas. The rats undergoing treatment received a saline solution or increasing concentrations of rBbCI-His(6), heparin, or a mixture of both. After 4 h of ligature VC, thrombus, if present was removed and weighed. aPTT, PT, and cytokines were measured in blood collected by cardiac puncture. aPTT, PT, and bleeding time (BT) were also measured at the time of VC (vena cava) ligature. rBbCI-His(6) (0.45 or 1.40 mg/kg) does not alter aPTT, PT or BT. No differences in coagulation parameters were detected in rBbCI-His(6) treated rats at the time of VC ligature or when the thrombus was removed. There was a significant decrease in the weight of thrombus in the animals of the groups treated with the rBbCI-His(6) (1.40 mg/kg), with the rBbCI-His(6) mixture (1.40 mg/kg) + heparin (50 IU/kg) and heparin (100 IU/kg) in relation to control group (saline). The growth-related oncogene/keratinocyte chemoattractant (GRO/KC) serum levels in rats treated with rBbCI-His(6) (1.40 mg/kg) or heparin (200 IU/kg) were reduced. In the experimental model used, rBbCI-His(6) alone had an antithrombotic effect, not altering blood clotting or bleeding time.

Highlights

Deep venous thrombosis (DVT) and its main complication, pulmonary thromboembolism, jointly with venous thromboembolism (VTE), constitute a global disease that is the third cause of cardiovascular deaths[1,2]
Since the 1960s, it is known that this flow reduction causes the formation of vortices at the level of valve sinus in leg veins. These events could lead to an accumulation of platelets that could be responsible for local activation of coagulation and accumulation of activated clotting factors, which would lead to thrombus formation and extension[7]
The native BbCI blocks the activity of neutrophil elastase, cathepsin G, and proteinase 3

Summary

Introduction

Deep venous thrombosis (DVT) and its main complication, pulmonary thromboembolism, jointly with venous thromboembolism (VTE), constitute a global disease that is the third cause of cardiovascular deaths[1,2]. Since the 1960s, it is known that this flow reduction causes the formation of vortices at the level of valve sinus in leg veins. These events could lead to an accumulation of platelets that could be responsible for local activation of coagulation and accumulation of activated clotting factors, which would lead to thrombus formation and extension[7]. Anticoagulants (heparin, low molecular heparins, fondaparinux, anti-vitamin K, and more recently direct oral anticoagulants) are used in the prophylaxis and treatment of VTE Efficient, they may present bleeding complications that can increase the morbidity and mortality of p atients[17,18]

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