Abstract
In our in vitro and in vivo studies, we used Acalypha indica root methanolic extract (AIRME), and investigated their free radical scavenging/antioxidant and anti-inflammatory properties. Primarily, phytochemical analysis showed rich content of phenols (70.92 mg of gallic acid/g) and flavonoids (16.01 mg of rutin/g) in AIRME. We then performed HR-LC-MS and GC-MS analyses, and identified 101 and 14 phytochemical compounds, respectively. Among them, ramipril glucuronide (1.563%), antimycin A (1.324%), swietenine (1.134%), quinone (1.152%), oxprenolol (1.118%), choline (0.847%), bumetanide (0.847%) and fenofibrate (0.711%) are the predominant phytomolecules. Evidence from in vitro studies revealed that AIRME scavenges DPPH and hydroxyl radicals in a concentration dependent manner (10–50 μg/mL). Similarly, hydrogen peroxide and lipid peroxidation were also remarkably inhibited by AIRME as concentration increases (20–100 μg/mL). In vitro antioxidant activity of AIRME was comparable to ascorbic acid treatment. For in vivo studies, carrageenan (1%, sub-plantar) was injected to rats to induce localized inflammation. Acute inflammation was represented by paw-edema, and significantly elevated (p < 0.05) WBC, platelets and C-reactive protein (CRP). However, AIRME pretreatment (150/300 mg/kg bodyweight) significantly (p < 0.05) decreased edema volume. This was accompanied by a significant (p < 0.05) reduction of WBC, platelets and CRP with both doses of AIRME. The decreased activities of superoxide dismutase, catalase, glutathione reductase and glutathione peroxidase in paw tissue were restored (p < 0.05 / p < 0.01) with AIRME in a dose-dependent manner. Furthermore, AIRME attenuated carrageenan-induced neutrophil infiltrations and vascular dilation in paw tissue. For the first time, our findings demonstrated the potent antioxidant and anti-inflammatory properties of AIRME, which could be considered to develop novel anti-inflammatory drugs.
Highlights
Inflammation is a complex biological immune response to harmful stimuli of irritant chemicals, pathogens and damaged local injury cells in the tissue
We quantified major phytochemicals, phenols and flavonoids based on qualitative analysis, which aided in further fractionation, isolation and biological assessment
The chromatograms obtained from Gas Chromatography-Mass Spectrometry (GC-MS) and HR-LC-MS analyses (Supplementary Materials Figures S1 and S2) and chemical structures of all compounds identified in A. indica root methanolic extract (AIRME) (Figures S3 and S4) were provided as Supplementary Materials
Summary
Inflammation is a complex biological immune response to harmful stimuli of irritant chemicals, pathogens and damaged local injury cells in the tissue. Inflammatory system plays an important role in life-threatening diseases including COVID-19, cancer, diabetes and cardiovascular dysfunction [1,2] Leukocyte activation such as macrophages, neutrophils, eosinophils and monocytes may occur during inflammation in order to protect the tissue, and simultaneously trigger the production of reactive oxygen species (ROS) or free radicals [3]. These ROS, include superoxide anion (O2−), hydrogen peroxide (H2O2), and hydroxyl radical (OH) are highly reactive in nature, which counteract with adjacent polyunsaturated fatty acids, proteins and DNA, and leading to cause lipid peroxidation, protein degradation, and DNA damage, respectively [4,5]. It is necessary to supplement anti-inflammatory and/or antioxidant compounds to assist the impaired immune and antioxidant defense systems
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