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Antioxidant and Antithrombotic Activities of Kenaf Seed (Hibiscus cannabinus) Coat Ethanol Extract in Sprague Dawley Rats

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Abstract

Oxidative stress has been implicated in deadly lifestyle diseases, and antioxidants from plant sources are the primary option in the treatment regime. Kenaf seeds are the storehouse of potential natural antioxidant phytoconstituents. Perhaps, none of the studies documented the phytoconstituents and their antioxidant potential from Kenaf seed coat so far. Thus, the current study focuses on exploring the protective effect of Kenaf Seed Coat Ethanol Extract (KSCEE) against sodium nitrite and diclofenac-induced oxidative stress in vitro (red blood cell and platelets model) and in vivo (female Sprague Dawely rat’s model) along with the antithrombotic activity. The infrared spectrophotometry data showed the heterogeneous functional groups (CH, OH, C = C, C = C–C) and aromatic rings. Reverse phase high-performance liquid chromatography and gas chromatography–mass spectrometry chromatogram of KSCEE also evidenced the presence of several phytochemicals. KSCEE displayed about 76% of DPPH scavenging activity with an IC50 value of 34.94 µg/ml. KSCEE significantly (***p < 0.001) normalized the stress markers such as lipid peroxidation, protein carbonyl content, superoxide dismutase, and catalase in sodium nitrite and diclofenac-induced oxidative stress in RBC, platelets, liver, kidney, and small intestine, respectively. Furthermore, KSCEE was found to protect the diclofenac-induced tissue destruction of the liver, kidney, and small intestine obtained from seven groups of female Sprague Dawely rats. KSCEE delayed the clotting time of platelet-rich plasma and platelet-poor plasma and activated partial thromboplastin time, suggesting its anticoagulant property. In addition, KSCEE also exhibited antiplatelet activity by inhibiting both adenosine diphosphate and epinephrine-induced platelet aggregation. In conclusion, KSCEE ameliorates the sodium nitrite and diclofenac-induced oxidative stress in red blood cells, platelets, and experimental animals along with antithrombotic properties.

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Data Availability

The data that support the finding of this study are available from the corresponding author upon reasonable request.

Abbreviations

ADP:

Adenosine diphosphate

ALP:

Alkaline phosphatase

APTT:

Activated partial thromboplastin time

CAT:

Catalase

DFC:

Diclofenac

DNPH:

Di-Nitro-Phenyl-Hydrazine

DPPH:

Di-Phenyl-2-Picryl-Hdrazyl

ECNICI:

Electron capture negative ion chemical ionization

EDTA:

Ethylene diamine tetraacetic acid

FTIR:

Fourier transform infrared spectrophotometry

GC–MS:

Gas chromatography-mass spectrometry

HPLC:

High-performance liquid chromatography

KSCEE:

Kenaf seed coat ethanol extract

LDH:

Lactate dehydrogenase

LPO:

Lipid per oxidation

MED:

Minimum edema dose

MHD:

Minimum hemorrhagic dose

PCC:

Protein carbonyl content

PDA:

Photometric diode array

PPP:

Platelet-poor plasma

PRP:

Platelet rich plasma

PT:

Prothrombin time

RBC:

Red blood cells

RNS:

Reactive nitrogen species

ROS:

Reactive oxygen species

SDS:

Sodium dodecyl sulfate

SGOT:

Serum glutamic oxaloacetic transamylase

SGPT:

Serum glutamate pyruvate transferase

SOD:

Super oxide dismutase

SYL:

Silymarin

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Acknowledgements

The authors thank Vision Group of Science and Technology, Government of Karnataka, India, for financial support and Liveon Biolabs Private Limited, Tumkur, Karnataka, India, for extending animal studies. The authors also thank the Department of Science and Technology (DST), Govt. of India, for providing the instrumentation facility under the scheme ‘Fund for Improvement of Science and Technology’ (Grant No. FIST-No. SR/FST/LS1/2018/175(C)) to the Department of Biochemistry, Kuvempu University Shankaraghatta, Karnataka, to achieve this piece of work and for awarding inspire fellowship to carry out the Ph.D. degree of the candidate.

Funding

This work is supported by Vision Group of Science and Technology, Government of Karnataka, India (VGST/CISEE/2012–13/282, dated March 16th, 2013). DST, New Delhi 2018/IF_180400.

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Authors and Affiliations

  1. Department of Biochemistry Jnansahydri, Kuvempu University, Shankarghatta-577451, Shivamogga, Karnataka, India

    Sujatha M. Hanumegowda, Manjula M.Venkatappa & Sathisha J. Gonchigar

  2. Department of Studies and Research in Biochemistry and Centre for Bioscience and Innovation, Tumkur University, B H road, Tumkur-572103, Tumkur, Karnataka, India

    Chandramma Srinivasa, Ashwini Shivaiah & Devaraja Sannaningaiah

  3. Department of Medical Biochemistry and Microbiology (IMBM), Uppsala Biomedical Centre, 75237, Uppsala, Sweden

    Sebestian Santhosh

  4. Liveon Biolabs Private Limited, Tumkur-572106, Tumkur, India

    Rajesh R

Authors
  1. Sujatha M. Hanumegowda
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  2. Chandramma Srinivasa
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  3. Ashwini Shivaiah
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  4. Manjula M.Venkatappa
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  5. Sebestian Santhosh
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  6. Rajesh R
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  7. Sathisha J. Gonchigar
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  8. Devaraja Sannaningaiah
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Contributions

SH and DS together planned and designed the research work. CR, AS, and MV assisted in the laboratory work. SSM, RR, and SG reviewed the article.

Corresponding authors

Correspondence to Sathisha J. Gonchigar or Devaraja Sannaningaiah.

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Ethics Approval

The research work involving only Sprague Dawley Female rats weighing 150–180 g was procured from Liveon Biolabs, limited. Institutional Animal ethical committee (TUMKUR, INDIA). All the experimentations were conducted under ethical guidelines and were approved by the Institutional Human Ethical Committee, Tumkur University, and Tumkur. Conducting animal experiments was permitted by the Institutional Animal Ethical Committee, Liveon Biolabs Private Limited, Tumkur. The animal handling was preceded following the guidelines of the Committee for Monitoring and Supervision of Experiments on Animals (CPCSEA). Ethical clearance was taken for this study from Liveon Biolabs, limited. Institutional Animal ethical committee (LBLIAEC) TUMKUR, with a protocol number LBPL-IAEC-47–05-2019.

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Hanumegowda, S.M., Srinivasa, C., Shivaiah, A. et al. Antioxidant and Antithrombotic Activities of Kenaf Seed (Hibiscus cannabinus) Coat Ethanol Extract in Sprague Dawley Rats. Appl Biochem Biotechnol 195, 772–800 (2023). https://doi.org/10.1007/s12010-022-04144-8

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