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<p>The post-COVID period has been accompanied by a notable rise in cardiovascular disease (CVD), emphasizing the importance of evaluating thrombolytic agents across heterogeneous populations. Although nattokinase (NK) is known for its fibrinolytic properties, systematic data on its clot dissolution efficiency and clot dissolution time (CDT) in the Indian population remain limited. Hence, this study examined NK-mediated thrombolysis across ABO/RhD blood groups, age strata, and gender supported by molecular docking and ultrastructural analyses. A total of 1,796 blood samples encompassing all eight ABO/RhD blood groups were collected from individuals aged 10–70 years from the Saurashtra region. In vitro clot lysis assays were used to quantify CDT following NK treatment, while scanning electron microscopy (SEM) and molecular docking &amp; dynamic were employed to investigate NK–fibrin–RhD interactions. Significant variation in CDT was observed across blood groups, with the longest dissolution time recorded in AB− females (2660&#xa0;s) and the shortest in O+ males (1510&#xa0;s). Four-way ANOVA analysis revealed a significant three-way interaction effect of age group x ABO blood group x RhD factor (<i>p</i> = 0.005), whereas the four-way interaction including gender was not significant (<i>p</i> = 1.000). CDT trends were further validated using an independent cohort of 562 samples collected from nine distinct locations across India, which showed consistent patterns. Molecular interaction analysis revealed that the RhD–NK–fibrin complex displayed substantially higher binding affinity (− 1813.9&#xa0;kcal/mol) than the NK–fibrin complex alone (− 733.3&#xa0;kcal/mol), aligned with enhanced clot dissolution efficiency observed in SEM analyses. In conclusion, NK-mediated CDT is not uniform across blood groups and is influenced by age and blood group antigen composition. These findings recommend large-scale in vivo validation studies to optimize NK dosing across diverse populations.</p>

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Discovery of age and blood group associated variability in nattokinase mediated thrombolysis and its relevance to cardiovascular management

  • 期刊论文

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  • 发表日期：

    2026年2月7日

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  • Tirth Chetankumar Bhatt,
  • Ashok Kumar Bishoyi

摘要

The post-COVID period has been accompanied by a notable rise in cardiovascular disease (CVD), emphasizing the importance of evaluating thrombolytic agents across heterogeneous populations. Although nattokinase (NK) is known for its fibrinolytic properties, systematic data on its clot dissolution efficiency and clot dissolution time (CDT) in the Indian population remain limited. Hence, this study examined NK-mediated thrombolysis across ABO/RhD blood groups, age strata, and gender supported by molecular docking and ultrastructural analyses. A total of 1,796 blood samples encompassing all eight ABO/RhD blood groups were collected from individuals aged 10–70 years from the Saurashtra region. In vitro clot lysis assays were used to quantify CDT following NK treatment, while scanning electron microscopy (SEM) and molecular docking & dynamic were employed to investigate NK–fibrin–RhD interactions. Significant variation in CDT was observed across blood groups, with the longest dissolution time recorded in AB− females (2660 s) and the shortest in O+ males (1510 s). Four-way ANOVA analysis revealed a significant three-way interaction effect of age group x ABO blood group x RhD factor (p = 0.005), whereas the four-way interaction including gender was not significant (p = 1.000). CDT trends were further validated using an independent cohort of 562 samples collected from nine distinct locations across India, which showed consistent patterns. Molecular interaction analysis revealed that the RhD–NK–fibrin complex displayed substantially higher binding affinity (− 1813.9 kcal/mol) than the NK–fibrin complex alone (− 733.3 kcal/mol), aligned with enhanced clot dissolution efficiency observed in SEM analyses. In conclusion, NK-mediated CDT is not uniform across blood groups and is influenced by age and blood group antigen composition. These findings recommend large-scale in vivo validation studies to optimize NK dosing across diverse populations.