Study on in vitro Thrombolytic Efficacy of Polyherbal Formulation to Mitigate Cardiovascular Risks after Recovery from COVID-19
Abstract
According to WHO reports, after recovering from COVID-19, approximately 10–20% of individuals experience a 1.8-fold higher risk of cardiovascular complications compared to non-infected individuals, even one year after recovery, due to hypercoagulability. This study aimed to develop a herbal remedy to mitigate cardiovascular risks in COVID-19 recovered individuals. An aqueous polyherbal formulation composed of the leaves of Ocimum sanctum, Moringa oleifera, Cardiospermum halicacabum, and the rhizome of Curcuma amada was prepared and subjected to phytochemical and spectral analysis. Blood samples from thirty COVID-19 recovered individuals were analysed for the in vitro thrombolytic efficacy and in vitro anti-platelet aggregation efficacy of the polyherbal formulation at different concentrations (8.2, 16.5, 33.0, and 66.0 mg/100 μl). Phytochemical screening revealed the presence of polyphenols, alkaloids, flavonoids, and saponins. Spectral analysis indicated the presence of phenolic O-H groups, conjugated C=O, and C=C bonds as functional groups. The polyherbal formulation demonstrated the highest thrombolytic activity of 79 ± 1.8% at 66 mg/100 μl compared to the standard streptokinase, and an anti-platelet aggregation activity of 83 ± 3.4% (IC₅₀ = 17.3 mg/100 μl) compared to standard aspirin. Hence, treatment with this polyherbal formulation may serve as a preventive therapy to mitigate cardiovascular risks following recovery from COVID-19.
Keywords:
Antiplatelet Efficacy, COVID-19-Post Complications, Polyherbal Remedy, Thrombolytic Activity
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References
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Barrett, T. J., Bilaloglu, S., Cornwell, M., Burgess, H. M., Virginio, V. W., Drenkova, K., ... & Berger, J. S. (2021). Platelets contribute to disease severity in COVID‐19. Journal of Thrombosis and Haemostasis, 19(12), 3139-3153. https://doi.org/10.1111/jth.15534
Bojić, M., Maleš, Ž., Antolić, A., Babić, I., & Tomičić, M. (2019). Antithrombotic activity of flavonoids and polyphenols rich plant species. Acta Pharmaceutica, 69(4), 483-495. https://doi.org/10.2478/acph-2019-0050
Chang, S., Yang, Z., Han, N., Liu, Z., & Yin, J. (2018). The antithrombotic, anticoagulant activity and toxicity research of ambinine, an alkaloid from the tuber of Corydalis ambigua var. amurensis. Regulatory Toxicology and Pharmacology, 95, 175-181. https://doi.org/10.1016/j.yrtph.2018.03.004
Cheng, H. L., Zhang, L. J., Liang, Y. H., Hsu, Y. W., Lee, I. J., Liaw, C. C., ... & Kuo, Y. H. (2013). Antiinflammatory and antioxidant flavonoids and phenols from Cardiospermum halicacabum (倒地鈴 Dào Dì Líng). Journal of Traditional and Complementary Medicine, 3(1), 33-40. https://doi.org/10.1016/s2225-4110(16)30165-1
Curry, S. J., Krist, A. H., Owens, D. K., Barry, M. J., Caughey, A. B., Davidson, K. W., ... & US Preventive Services Task Force. (2018). Risk assessment for cardiovascular disease with nontraditional risk factors: US preventive services task force recommendation statement. Jama, 320(3), 272-280. https://doi.org/10.1001/jama.2018.8359
Ha, H. A., Al-Humaid, L. A., Aldawsari, M., Bharathi, D., & Lee, J. (2024). Evaluation of phytochemical, antibacterial, thrombolytic, anti-inflammatory, and cytotoxicity profile of Achyranthes aspera aerial part extracts. Environmental Research, 243, 117802. https://doi.org/10.1016/j.envres.2023.117802
Harborne, A. J. (1998). Phytochemical methods a guide to modern techniques of plant analysis. springer science & business media. https://books.google.co.in/books?hl=en&lr=&id=2yvqeRtE8CwC&oi=fnd&pg=PR7&dq=Harborne,+J.+B.+(2012).+Phytochemical+Methods:+A+Guide+to+Modern+Techniques+of+Plant+Analysis.+Springer+Science+%26+Business+Media.&ots=xBfoQ6SqU2&sig=cu2hf-K5t0gsZvTA-lGL3FINwUk&redir_esc=y#v=onepage&q&f=false
Helmy, S. A., Morsy, N. F., Elaby, S. M., & Ghaly, M. A. (2017). Hypolipidemic effect of Moringa oleifera Lam leaf powder and its extract in diet-induced hypercholesterolemic rats. Journal of Medicinal Food, 20(8), 755-762. https://doi.org/10.1089/jmf.2016.0155
Hottz, E. D., Azevedo-Quintanilha, I. G., Palhinha, L., Teixeira, L., Barreto, E. A., Pão, C. R., ... & Bozza, P. T. (2020). Platelet activation and platelet-monocyte aggregate formation trigger tissue factor expression in patients with severe COVID-19. Blood, The Journal of the American Society of Hematology, 136(11), 1330-1341. https://doi.org/10.1182/blood.2020007252
Hussain, F., Islam, A., Bulbul, L., Moghal, M. R., & Hossain, M. S. (2014). In vitro thrombolytic potential of root extracts of four medicinal plants available in Bangladesh. Ancient Science of Life, 33(3), 162-164. https://doi.org/10.4103/0257-7941.144620
Jain, V., Kunwar, B., & Verma, S. K. (2023). A review on thrombolysis enhancing indian edible plants. Biomedical and Pharmacology Journal, 16(3), 1283-1302. https://doi.org/10.13005/bpj/2709
Jing, H., Wu, X., Xiang, M., Liu, L., Novakovic, V. A., & Shi, J. (2022). Pathophysiological mechanisms of thrombosis in acute and long COVID-19. Frontiers in Immunology, 13, 992384. https://doi.org/10.3389/fimmu.2022.992384
Khan, I. N., Habib, M. R., Rahman, M. M., Mannan, A., Sarker, M. M. I., & Hawlader, S. (2011). Thrombolytic potential of Ocimum sanctum L., Curcuma longa L., Azadirachta indica L. and Anacardium occidentale L. Journal of Basic and Clinical Pharmacy, 2(3), 125. https://www.jbclinpharm.org/articles/thrombolytic-potential-of-ocimum-sanctum-l-curcuma-longa-l-azadirachta-indica-l-and-anacardium-occidentale-l.html
Kim, J. S. (2020). Posterior Circulation Stroke: Advances in Understanding and Management. Springer Nature.
Knight, R., Walker, V., Ip, S., Cooper, J. A., Bolton, T., Keene, S., Denholm, R., Akbari, A., Abbasizanjani, H., Torabi, F., Omigie, E., Hollings, S., North, T.-L., Toms, R., Di Angelantonio, E., Denaxas, S., Thygesen, J. H., Tomlinson, C., Bray, B., … Sterne, J. A. C. (2022). Association of COVID-19 with major arterial and venous thrombotic diseases: A population-wide cohort study of 48 million adults in England and Wales. Circulation, 146(12). 892-906. https://doi.org/10.1161/circulationaha.122.060785
Kunwar, B., Jain, V., & Verma, S. K. (2022). In vitro thrombolytic activity of Moringa oleifera. Nusantara Bioscience, 14(1). https://doi.org/10.13057/nusbiosci/n140108
Lin, J., Yan, H., Chen, H., He, C., Lin, C., He, H., ... & Lin, K. (2021). COVID‐19 and coagulation dysfunction in adults: A systematic review and meta‐analysis. Journal of Medical Virology, 93(2), 934-944. https://doi.org/10.1002/jmv.26346
Merschel, M. (2022). Blood clot risk remains elevated nearly a year after COVID-19. American Heart Association News. https://www.heart.org/en/news/2022/09/19/blood-clot-risk-remains-elevated-nearly-a-year-after-covid-19
S, V. B., V, A., Ahmad, A., & Tajo, S. M. (2017). Thrombolytic Activity of C. amada and C.caesia. Asian Journal of Pharmaceutical and Clinical Research, 10(2), 317–318. https://doi.org/10.22159/ajpcr.2017.v10i2.15694
Seyed Alinaghi, S., Afsahi, A. M., MohsseniPour, M., Behnezhad, F., Salehi, M. A., Barzegary, A., ... & Dadras, O. (2021). Late complications of COVID-19; a systematic review of current evidence. Archives of Academic Emergency Medicine, 9(1), e14. https://doi.org/10.22037/aaem.v9i1.1058
Srilatha, M., Aroulmoji, V., Rathinavel, T., Roumana, C., Hariharan, M., Bhuvaneswari, M., ... & Raja, R. K. Exploration and validation of thrombolytic phytocompounds from curcuma amada roxb through in vitro and in silico approaches. Medicinal Plant, 4, 5. https://doi.org/10.29294/IJASE.11.4.2025.4679-4702.
Tabassum, F., Chandi, S. H., Mou, K. N., Hasif, K., Ahamed, T., & Akter, M. (2017). Invitro thrombolytic activity and phytochemical evaluation of leaf extracts of four medicinal plants of Asteraceae family. Journal of Pharmacognosy and Phytochemistry, 6(4), 1166-1169. https://www.phytojournal.com/archives/2017/vol6issue4/PartQ/6-4-135-559.pdf
Tohti, I., Tursun, M., Umar, A., Turdi, S., Imin, H., & Moore, N. (2006). Aqueous extracts of Ocimum basilicum L. (sweet basil) decrease platelet aggregation induced by Sanger ADP and thrombin in vitro and rats arterio–venous shunt thrombosis in vivo. Thrombosis Research, 118(6), 733-739. https://doi.org/10.1016/j.thromres.2005.12.011
von Meijenfeldt, F. A., Havervall, S., Adelmeijer, J., Lundström, A., Magnusson, M., Mackman, N., ... & Lisman, T. (2021). Sustained prothrombotic changes in COVID-19 patients 4 months after hospital discharge. Blood advances, 5(3), 756-759. https://doi.org/10.1182/bloodadvances.2020003968
Williamson, L. (2024). COVID-19 may increase heart attack and stroke risk for years American Heart Association News. https://www.heart.org/en/news/2024/10/09/covid-19-may-increase-heart-attack-and-stroke-risk-for-years
Wu, Q. L., Dong, J., Zeng, H. W., Lv, C., Liu, A. J., & Zhang, W. D. (2020). Monitoring antiplatelet aggregation in vivo and in vitro by microtiter plate method. Journal of Cardiovascular Pharmacology, 75(4), 314-320. https://doi.org/10.1097/fjc.0000000000000801
Xie, Y., Xu, E., Bowe, B., & Al-Aly, Z. (2022). Long-term cardiovascular outcomes of COVID-19. Nature medicine, 28(3), 583-590. https://doi.org/10.1038/s41591-022-01689-3
Zhu, L. (2018). Tannic acid inhibits protein disulfide isomerase, platelet activation and thrombus formation. Blood, 132(Supplement 1), 2418. https://doi.org/10.1182/blood-2018-99-114418
Amrani, S., Harnafi, H., Gadi, D., Mekhfi, H., Legssyer, A., Aziz, M., ... & Bosca, L. (2009). Vasorelaxant and anti-platelet aggregation effects of aqueous Ocimum basilicum extract. Journal of Ethnopharmacology, 125(1), 157-162. https://doi.org/10.1016/j.jep.2009.05.043
Barrett, T. J., Bilaloglu, S., Cornwell, M., Burgess, H. M., Virginio, V. W., Drenkova, K., ... & Berger, J. S. (2021). Platelets contribute to disease severity in COVID‐19. Journal of Thrombosis and Haemostasis, 19(12), 3139-3153. https://doi.org/10.1111/jth.15534
Bojić, M., Maleš, Ž., Antolić, A., Babić, I., & Tomičić, M. (2019). Antithrombotic activity of flavonoids and polyphenols rich plant species. Acta Pharmaceutica, 69(4), 483-495. https://doi.org/10.2478/acph-2019-0050
Chang, S., Yang, Z., Han, N., Liu, Z., & Yin, J. (2018). The antithrombotic, anticoagulant activity and toxicity research of ambinine, an alkaloid from the tuber of Corydalis ambigua var. amurensis. Regulatory Toxicology and Pharmacology, 95, 175-181. https://doi.org/10.1016/j.yrtph.2018.03.004
Cheng, H. L., Zhang, L. J., Liang, Y. H., Hsu, Y. W., Lee, I. J., Liaw, C. C., ... & Kuo, Y. H. (2013). Antiinflammatory and antioxidant flavonoids and phenols from Cardiospermum halicacabum (倒地鈴 Dào Dì Líng). Journal of Traditional and Complementary Medicine, 3(1), 33-40. https://doi.org/10.1016/s2225-4110(16)30165-1
Curry, S. J., Krist, A. H., Owens, D. K., Barry, M. J., Caughey, A. B., Davidson, K. W., ... & US Preventive Services Task Force. (2018). Risk assessment for cardiovascular disease with nontraditional risk factors: US preventive services task force recommendation statement. Jama, 320(3), 272-280. https://doi.org/10.1001/jama.2018.8359
Ha, H. A., Al-Humaid, L. A., Aldawsari, M., Bharathi, D., & Lee, J. (2024). Evaluation of phytochemical, antibacterial, thrombolytic, anti-inflammatory, and cytotoxicity profile of Achyranthes aspera aerial part extracts. Environmental Research, 243, 117802. https://doi.org/10.1016/j.envres.2023.117802
Harborne, A. J. (1998). Phytochemical methods a guide to modern techniques of plant analysis. springer science & business media. https://books.google.co.in/books?hl=en&lr=&id=2yvqeRtE8CwC&oi=fnd&pg=PR7&dq=Harborne,+J.+B.+(2012).+Phytochemical+Methods:+A+Guide+to+Modern+Techniques+of+Plant+Analysis.+Springer+Science+%26+Business+Media.&ots=xBfoQ6SqU2&sig=cu2hf-K5t0gsZvTA-lGL3FINwUk&redir_esc=y#v=onepage&q&f=false
Helmy, S. A., Morsy, N. F., Elaby, S. M., & Ghaly, M. A. (2017). Hypolipidemic effect of Moringa oleifera Lam leaf powder and its extract in diet-induced hypercholesterolemic rats. Journal of Medicinal Food, 20(8), 755-762. https://doi.org/10.1089/jmf.2016.0155
Hottz, E. D., Azevedo-Quintanilha, I. G., Palhinha, L., Teixeira, L., Barreto, E. A., Pão, C. R., ... & Bozza, P. T. (2020). Platelet activation and platelet-monocyte aggregate formation trigger tissue factor expression in patients with severe COVID-19. Blood, The Journal of the American Society of Hematology, 136(11), 1330-1341. https://doi.org/10.1182/blood.2020007252
Hussain, F., Islam, A., Bulbul, L., Moghal, M. R., & Hossain, M. S. (2014). In vitro thrombolytic potential of root extracts of four medicinal plants available in Bangladesh. Ancient Science of Life, 33(3), 162-164. https://doi.org/10.4103/0257-7941.144620
Jain, V., Kunwar, B., & Verma, S. K. (2023). A review on thrombolysis enhancing indian edible plants. Biomedical and Pharmacology Journal, 16(3), 1283-1302. https://doi.org/10.13005/bpj/2709
Jing, H., Wu, X., Xiang, M., Liu, L., Novakovic, V. A., & Shi, J. (2022). Pathophysiological mechanisms of thrombosis in acute and long COVID-19. Frontiers in Immunology, 13, 992384. https://doi.org/10.3389/fimmu.2022.992384
Khan, I. N., Habib, M. R., Rahman, M. M., Mannan, A., Sarker, M. M. I., & Hawlader, S. (2011). Thrombolytic potential of Ocimum sanctum L., Curcuma longa L., Azadirachta indica L. and Anacardium occidentale L. Journal of Basic and Clinical Pharmacy, 2(3), 125. https://www.jbclinpharm.org/articles/thrombolytic-potential-of-ocimum-sanctum-l-curcuma-longa-l-azadirachta-indica-l-and-anacardium-occidentale-l.html
Kim, J. S. (2020). Posterior Circulation Stroke: Advances in Understanding and Management. Springer Nature.
Knight, R., Walker, V., Ip, S., Cooper, J. A., Bolton, T., Keene, S., Denholm, R., Akbari, A., Abbasizanjani, H., Torabi, F., Omigie, E., Hollings, S., North, T.-L., Toms, R., Di Angelantonio, E., Denaxas, S., Thygesen, J. H., Tomlinson, C., Bray, B., … Sterne, J. A. C. (2022). Association of COVID-19 with major arterial and venous thrombotic diseases: A population-wide cohort study of 48 million adults in England and Wales. Circulation, 146(12). 892-906. https://doi.org/10.1161/circulationaha.122.060785
Kunwar, B., Jain, V., & Verma, S. K. (2022). In vitro thrombolytic activity of Moringa oleifera. Nusantara Bioscience, 14(1). https://doi.org/10.13057/nusbiosci/n140108
Lin, J., Yan, H., Chen, H., He, C., Lin, C., He, H., ... & Lin, K. (2021). COVID‐19 and coagulation dysfunction in adults: A systematic review and meta‐analysis. Journal of Medical Virology, 93(2), 934-944. https://doi.org/10.1002/jmv.26346
Merschel, M. (2022). Blood clot risk remains elevated nearly a year after COVID-19. American Heart Association News. https://www.heart.org/en/news/2022/09/19/blood-clot-risk-remains-elevated-nearly-a-year-after-covid-19
S, V. B., V, A., Ahmad, A., & Tajo, S. M. (2017). Thrombolytic Activity of C. amada and C.caesia. Asian Journal of Pharmaceutical and Clinical Research, 10(2), 317–318. https://doi.org/10.22159/ajpcr.2017.v10i2.15694
Seyed Alinaghi, S., Afsahi, A. M., MohsseniPour, M., Behnezhad, F., Salehi, M. A., Barzegary, A., ... & Dadras, O. (2021). Late complications of COVID-19; a systematic review of current evidence. Archives of Academic Emergency Medicine, 9(1), e14. https://doi.org/10.22037/aaem.v9i1.1058
Srilatha, M., Aroulmoji, V., Rathinavel, T., Roumana, C., Hariharan, M., Bhuvaneswari, M., ... & Raja, R. K. Exploration and validation of thrombolytic phytocompounds from curcuma amada roxb through in vitro and in silico approaches. Medicinal Plant, 4, 5. https://doi.org/10.29294/IJASE.11.4.2025.4679-4702.
Tabassum, F., Chandi, S. H., Mou, K. N., Hasif, K., Ahamed, T., & Akter, M. (2017). Invitro thrombolytic activity and phytochemical evaluation of leaf extracts of four medicinal plants of Asteraceae family. Journal of Pharmacognosy and Phytochemistry, 6(4), 1166-1169. https://www.phytojournal.com/archives/2017/vol6issue4/PartQ/6-4-135-559.pdf
Tohti, I., Tursun, M., Umar, A., Turdi, S., Imin, H., & Moore, N. (2006). Aqueous extracts of Ocimum basilicum L. (sweet basil) decrease platelet aggregation induced by Sanger ADP and thrombin in vitro and rats arterio–venous shunt thrombosis in vivo. Thrombosis Research, 118(6), 733-739. https://doi.org/10.1016/j.thromres.2005.12.011
von Meijenfeldt, F. A., Havervall, S., Adelmeijer, J., Lundström, A., Magnusson, M., Mackman, N., ... & Lisman, T. (2021). Sustained prothrombotic changes in COVID-19 patients 4 months after hospital discharge. Blood advances, 5(3), 756-759. https://doi.org/10.1182/bloodadvances.2020003968
Williamson, L. (2024). COVID-19 may increase heart attack and stroke risk for years American Heart Association News. https://www.heart.org/en/news/2024/10/09/covid-19-may-increase-heart-attack-and-stroke-risk-for-years
Wu, Q. L., Dong, J., Zeng, H. W., Lv, C., Liu, A. J., & Zhang, W. D. (2020). Monitoring antiplatelet aggregation in vivo and in vitro by microtiter plate method. Journal of Cardiovascular Pharmacology, 75(4), 314-320. https://doi.org/10.1097/fjc.0000000000000801
Xie, Y., Xu, E., Bowe, B., & Al-Aly, Z. (2022). Long-term cardiovascular outcomes of COVID-19. Nature medicine, 28(3), 583-590. https://doi.org/10.1038/s41591-022-01689-3
Zhu, L. (2018). Tannic acid inhibits protein disulfide isomerase, platelet activation and thrombus formation. Blood, 132(Supplement 1), 2418. https://doi.org/10.1182/blood-2018-99-114418
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How to Cite
B., V., Firdous, J., Anand, V. and M., A. (2025) “Study on in vitro Thrombolytic Efficacy of Polyherbal Formulation to Mitigate Cardiovascular Risks after Recovery from COVID-19”, International Journal of Advancement in Life Sciences Research, 8(4), pp. 1-10. doi: https://doi.org/10.31632/ijalsr.2025.v08i04.001.
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