Protection of mouse brain from paracetamol-induced stress by Centella asiatica methanol extract

• Published in: Journal of ethnopharmacology 2019-05, Vol.236, p.474-483
• Main Authors: Viswanathan, Gayathri, Dan, Vipin Mohan, Radhakrishnan, Neelima, Nair, Akhila Sasikumar, Rajendran Nair, Aroma Prasanna, Baby, Sabulal
• Format: Article
• Language: English
• Subjects: Acetaminophen - toxicity; Animals; Anti-inflammatory; Antioxidant; Apoptosis; Astrocytes; Astrocytes - drug effects; Astrocytes - pathology; Behavior, Animal - drug effects; Brain; Brain - cytology; Brain - drug effects; Brain - pathology; Cells, Cultured; Centella - chemistry; Centella asiatica; Disease Models, Animal; Drug Evaluation, Preclinical; Drug Overdose - complications; Drug Overdose - drug therapy; Drug Overdose - etiology; Ethnomedicine; Humans; Inflammation - drug therapy; Inflammation - etiology; Inflammation - pathology; Male; Medicine, Ayurvedic; Methanol - chemistry; Mice; Neuroprotective Agents - isolation & purification; Neuroprotective Agents - pharmacology; Neuroprotective Agents - therapeutic use; Oxidative Stress - drug effects; Plant Extracts - isolation & purification; Plant Extracts - pharmacology; Plant Extracts - therapeutic use; Primary Cell Culture; Triterpenes - isolation & purification; Triterpenes - pharmacology
• ISSN: 0378-8741; 1872-7573
• DOI: 10.1016/j.jep.2019.03.017

Centella asiatica (CA) is a medicinal herb traditionally used as a brain tonic in Ayurvedic medicine. Various ethnomedical leads revealed the effective use of CA in the treatment of symptoms associated to oxidative stress and inflammation. The aim of this study was to evaluate the therapeutic ability of CA methanol extract (CAM) in protecting mouse brain and astrocytes from oxidative stress and inflammation induced by Paracetamol, and thus to substantiate the allied traditional/ethnomedical claims of CA. Chemical profiling of CAM and quantification of its major constituents were carried out by HPTLC-densitometry. Mice were administered with CAM and Paracetamol in various combinations, and oxidative stress parameters (lipid peroxidation, radical scavenging) as well as nitric oxide stress were estimated from isolated mouse brain. Cellular toxicity was investigated by apoptosis/necrosis in primary astrocytes isolated from brain tissues of mouse (which was challenged by CAM/Paracetamol) by flow cytometry and fluorescent microscopy. Expression of inflammatory cytokine mediators (monocyte chemo attractant protein 1, interleukin 1, interferon γ, tumor necrosis factor β, interleukin 10 and mitogen activated protein kinase 14 gene) in CAM/Paracetamol administered mouse brain tissues was analyzed by real time PCR. Mouse brain tissues challenged by CAM/Paracetamol were also assessed for gross and histopathology. In addition, staining with acridine orange was carried out in C6 cell lines treated with CAM, and viewed under fluorescent microscopy. Paracetamol elicited reactive oxygen species generation was revealed through Ferric Reducing Antioxidant Power (FRAP) activity. CAM reversed the Paracetamol induced free radical and reactive nitrogen species production and increased the scavenging activity which was more pronounced at the higher dose (80 mg/kg b.wt). CAM negated the Paracetamol-induced damage by inhibiting expression of pro-inflammatory cytokines (MCP 1, IL 1, TNF β), and increasing the expression of the anti-inflammatory cytokine (IL 10) profoundly. Interestingly, MAPK 14 gene expression was decreased gradually and became same as normal control with increase in the dose of CAM. Also, it was evident that CAM protected mouse primary astrocytes from Paracetamol by maintaining a normal morphology. Similarly, apoptosis of primary astrocytes (treated with Paracetamol/CAM) decreased with the increase in CAM dose (80 mg/kg b.wt.) which was evident from flow cytometri