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KMID : 1200820140140030255
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Oriental Pharmacy and Experimental Medicine
2014 Volume.14 No. 3 p.255 ~ p.268
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In-vitro cytoprotective activity of Smilax zeylanica leaves against hydrogen peroxide induced oxidative stress in L-132 and BRL 3A cells
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Rajesh Venugopalan
Perumal Perumal
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Abstract
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The aim of this study was to investigate the cytoprotective effect of different solvent extracts of Smilax zeylanica leaves against hydrogen peroxide induced oxidative stress in L132 pulmonary cells and BRL 3A liver cells. Powdered leaves of Smilax zeylanica were defatted with petroleum ether and subjected to successive extraction in soxhlet extractor using solvents of increasing polarity (benzene, chloroform and methanol). Initially, the solvent extracts were subjected to qualitative and quantitative analysis and assessed for its potential protective effect against hydrogen peroxide (H2O2) induced oxidative stress in L132 and BRL 3A cell lines. Some biochemical assays were carried out to determine the cytoprotective activity, including cell viability, lipid peroxidation by determining the formation of malondialdehyde, lactate dehydrogenase leakage into culture medium, the catalase activity and the content of reduced glutathione (GSH) in the cells. Exposure of L 132 and BRL 3A to 2 mM H2O2, reduced the cell viability, increased the malondialdehyde (MDA) level, increased the leakage of lactate dehydrogenase (LDH) and caused reduction in antioxidant activities. It was observed that pretreatment of cultured cells with methanol extract of Smilax zeylanica leaves exhibited a significant cytoprotection by increasing the cell viability, decreasing lipid peroxidation, LDH leakage and increasing the catalase and reduced glutathione content in the cells. These findings suggest that methanol extract of Smilax zeylanica leaves has a strong cytoprotective activity against oxidative injury caused by reactive oxygen species. GC-MS analysis was carried out to determine the possible chemical components from methanol extract of Smilax zeylanica leaves. The analysis revealed the presence of phytoconstituents hydroxytyrosol, trans-iso-eugenol and squalene. Hydroxytyrosol is believed to be one of the most powerful antioxidants. Trans-iso-eugenol is reported to have potent antioxidant activity. In addition, squalene compound is reported as an antioxidant, antitumor, cancer-preventive.
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KEYWORD
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Smilax zeylanica leaves, Hydrogen peroxide, Oxidative stress, In-vitro cytoprotective, Human pulmonary cells, Rat liver cells, GC-MS analysis
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