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Oxidative Stress-Induced Platelet Apoptosis/Activation: Alleviation by Purified Curcumin via ASK1-JNK/p-38 Pathway

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Abstract

Platelets are known for their indispensable role in hemostasis and thrombosis. However, alteration in platelet function due to oxidative stress is known to mediate various health complications, including cardiovascular diseases and other health complications. To date, several synthetic molecules have displayed antiplatelet activity; however, their uses are associated with bleeding and other adverse effects. The commercially available curcumin is generally a mixture of three curcuminoids: curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Although crude curcumin is known to inhibit platelet aggregation, the effect of purified curcumin on platelet apoptosis, activation, and aggregation remains unclear. Therefore, in this study, curcumin was purified from a crude curcumin mixture and the effects of this preparation on the oxidative stress-induced platelet apoptosis and activation was evaluated. 2,2′-Azobis(2-methylpropionamidine) dihydrochloride (AAPH) compound was used as an inducer of oxidative stress. Purified curcumin restored AAPH-induced platelet apoptotic markers like reactive oxygen species, intracellular calcium level, mitochondrial membrane potential, cardiolipin peroxidation, cytochrome c release from mitochondria to the cytosol, and phosphatidyl serine externalization. Further, it inhibited the agonist-induced platelet activation and aggregation, demonstrating its antiplatelet activity. Western blot analysis confirms protective effect of the purified curcumin against oxidative stress-induced platelet apoptosis and activation via downregulation of MAPKs protein activation, including ASK1, JNK, and p-38. Together, these results suggest that the purified curcumin could be a potential therapeutic bioactive molecule to treat the oxidative stress-induced platelet activation, apoptosis, and associated complications.

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Abbreviations

AAPH:

2,2′-Azobis(2-methylpropionamidine) dihydrochloride

BDMC:

bisdemethoxycurcumin

CVD:

cardiovascular disease

DMC:

demethoxycurcumin

GSH:

glutathione

ROS:

reactive oxygen species

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Acknowledgments

KM thanks ICMR-SRF (File No. 45/25/2020-BIO/BMS and Dated 10.3.2021) for the research fellowship. Authors thank Central instrumentation facility, Institution of excellence (IOE), the University of Mysore for providing central instrumentation facilities. K.K. acknowledges funding from the DST-SERB (EEQ/2021/000142). KSG acknowledges funding from the DST-SERB (EEQ/2021/000375). The authors thanks members of the KK lab for their kind help during the study.

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Authors and Affiliations

  1. Department of Studies in Biochemistry, University of Mysore, 570006, Mysuru, India

    Kurnegala Manikanta, Manoj Paul, Vaddaragudisalu D. Sandesha & Kempaiah Kemparaju

  2. Department of Biological Sciences, School of Dental Medicine, Case Western Reserve University, 44106, Cleveland, OH, USA

    Shanmuga S. Mahalingam

  3. Department of Studies and Research in Chemistry, University College of Science, Tumkur University, 572103, Tumakuru, India

    Thimmasandra Narayan Ramesh

  4. Department of Biotechnology, Yuvaraja’s College, University of Mysore, 570005, Mysore, India

    Krishnegowda Harishkumar

  5. All India Institute of Medical Science, 110029, New Delhi, India

    Shashank S. Koundinya

  6. Applied Nutrition Discipline, Defense Food Research Laboratory, 570011, Mysuru, India

    Shivanna Naveen

  7. Department of Studies and Research in Biochemistry, Tumkur University, 572103, Tumakuru, India

    Kesturu S. Girish

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Contributions

K.S.G., K.K., and M.P. designed the study; M.P. and K.M. performed the experiments; M.P., K.M., V.D.S., S.S.M., T.N.R., H.K., S.S.K., and S.N. analyzed the data; M.P., S.S.M., K.S.G., and K.K. wrote and edited the manuscript. K.S.G. and K.K. conceived and directly supervised the study. All authors have read and agreed to the published version the manuscript.

Corresponding authors

Correspondence to Kempaiah Kemparaju or Kesturu S. Girish.

Ethics declarations

Blood was drawn from the healthy human volunteers with informed consent, according to the Institutional Human Ethical Committee (IHEC-UOM No. 70/Res NOC/2020-21) guidelines, University of Mysore, Mysuru. The authors of this work declare that they have no conflicts of interest.

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Manikanta, K., Paul, M., Sandesha, V.D. et al. Oxidative Stress-Induced Platelet Apoptosis/Activation: Alleviation by Purified Curcumin via ASK1-JNK/p-38 Pathway. Biochemistry Moscow 89, 417–430 (2024). https://doi.org/10.1134/S0006297924030039

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