Sorghum Protein Extract Protects RBC from Sodium Nitrite-Induced Oxidative Stress and Exhibits Anticoagulant and Antiplatelet Activity

Devaraja Sannaningaiah,Ramesh Komalapura Laxmaiah,Sharath Kumar M Nandish,Ashwini Shivaiah,Chandramma Srinivasa,Bhagyalakshmi Manjappa,Chethana Ramachandraiah,Manohar Shinde,Jayanna Kengaiah,Sebastin Santosh Martin,Sujatha Hanumegowda

doi:10.3897/folmed.63.e57713

Devaraja Sannaningaiah, Ramesh Komalapura Laxmaiah + Show 9 more

Open Access

https://doi.org/10.3897/folmed.63.e57713

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Journal: Folia medica	Publication Date: Dec 31, 2021
Citations: 5	License type: CC BY 4.0

Affiliation: Tumkur University, Kuvempu University

Abstract

Oxidative stress plays a critical role in the progression of diabetes, arthritis, cancer, eryptosis, cardiovascular disease, and thrombosis. Currently, antioxidants from natural sources are in high demand due to their beneficial role in the management of said diseases. The purpose of the study was to evaluate the protective effect of sorghum protein buffer extract (SBE) on sodium nitrite-induced oxidative stress and thrombosis. Protein characterization of SBE was done using SDS-PAGE. Oxidative stress in RBC was induced using sodium nitrite (NaNO2) and the key stress markers such as lipid peroxidation (LPO), protein carbonyl content (PCC), and the level of antioxidant enzymes (SOD and CAT) were measured. The anticoagulant effect of SBE was identified by employing in-vitro plasma recalcification time, activated partial thromboplastin time (APTT), prothrombin time (PT), and in-vivo mouse tail bleeding time. SBE antiplatelet activity was examined using agonist adenosine diphosphate (ADP) and epinephrine-induced platelet aggregation. Non-toxic property of SBE was identified using in-vitro direct haemolytic, haemorrhagic, and edema forming activities using experimental mice. SBE revealed similar protein banding pattern under both reduced and non-reduced conditions on SDS-PAGE. Interestingly, SBE normalized the level of LPO, PCC, SOD, and CAT in stress-inducedRBCs. Furthermore, SBE showed anticoagulant effect in platelet rich plasma by enhancing the clotting time from the control 250 s to 610 s and bleeding time from the control 200 s to more than 500 s (p<0.01) in a dose dependent manner. In addition, SBE prolonged the clot formation process of only APTT but not PT. SBE inhibited the agonists ADP and epinephrine induced platelet aggregation. SBE did not hydrolyze RBC cells, devoid of edema and haemorrhage properties. This study demonstrates for the first time the anticoagulant, antiplatelet, and antioxidant properties of SBE. Thus, the observed results validate consumption of sorghum as good for health and well-being.

Highlights

Oxidative stress plays a critical role in the progression of diabetes, arthritis, cancer, eryptosis, cardiovascular disease, and thrombosis
Trichloroacetic acid (TCA), 95% ethanol, sodium nitrite (NaNO2), sodium dodecyl sulfate (SDS), acetic acid, thiobarbituric acid, dinitrophenyl hydrazine (DNPH), tetramethyl ethylenediamine (TEMED), ethylenediaminetetraacetic acid (EDTA), quercetin, and hydrogen peroxide were purchased from Sigma Chemicals Company
sorghum protein buffer extract (SBE) showed similar kind of protein bands from the range of 200 kDa to 18.4 kDa, suggesting the proteins present are monomeric in nature (Fig. 1)

Summary

Introduction

Oxidative stress plays a critical role in the progression of diabetes, arthritis, cancer, eryptosis, cardiovascular disease, and thrombosis. The establishment of novel antioxidant treatment remedies that target to avoid the thrombus formation and permits ROS modulation constrained to their physiological redox mechanisms.[8] Sorghum belongs to the family Poaceae, commonly called great millet/Indian millet.[9] Sorghum grain is the fifth most important cereal crop in the world after wheat, rice, corn, and barley.[10] It is the third most widely grown crop in India after rice and wheat It is the most important staple food for a large proportion of the population as well.[11] Sorghum is generally more economical to produce and used in a variety of foods.[12] The white food sorghums are processed into flour and other products including expanded snacks, cookies and ethnic foods, and are gaining popularity in Japan.[13] Whole grain consumption has been associated with a decreased incidence of cancer, cardiovascular disease (CVD), diabetes and obesity.[14] Sorghum is the richest source of macro (proteins, lipids and carbohydrates) and micro molecules (vitamins and minerals). The therapeutic usage of proteins from sorghum has been least explored

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