Neuroprotective Effect of Convolvulus pluricaulis Methanol Extract on Hydrogen Peroxide Induced Oxidative Stress in Human IMR32 Neuroblastoma Cell Line

Abstract

Aims: The present study aimed to evaluate and ascertain the protective role of methanolic/ethanolic/water extracts of Convolvulus pluricaulis against H2O2 induced cytotoxicity in IMR32 Neuroblastoma cell line as model system and identify the factor responsible for the protective effect. Study Design:  Experimental study. Place and Duration of Study: Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar & Department of Biotechnology, DAV College, Amritsar, PuCPab, between August 2010 and March 2012. Methodology: Firstly, cytotoxic dose of H2O2 ­and non-toxic dose of methanolic, ethanolic and water extracts of C. pluricaulis (CP-MEx, CP-EEx and CP-WEx respectively) was determined by MTT assay. Protective effect of CP-MEx, CP-EEx and CP-WEx was determined using quercetin as a positive control. The expression of IMR32 cytoskeletal marker, Neurofilament (NF-200) and stress markers, Heat shock protein (HSP70) and (glucose regulated protein 75, Grp75) Mortalin studied by immunofluorescence and RT-PCR results. The level of antioxidant enzymes catalase, superoxide dismutase, glutathione peroxidase, direct scavenger of free radicals, Glutathione and lipid peroxidation were analysed by their standard procedures. Results: The results showed that quercetin, CP-MEx, CP-EEx and CP-WEx displayed cytoprotective activity in IMR32 cells. Out of tested extracts CP-MEx significantly decreased hydrogen peroxide-induced cell death. Significant decrease in NF-200, HSP70 and Mortalin expression was observed in CP-MEx+H2O2 treated cultures as compared to H2O2 treated. Catalase, superoxide dismutase, glutathione peroxidase, Glutathione levels significantly increased in Quercetin and CP-MEx treated cultures. Lipid peroxidation was significantly decreased in both Quercetin and CP-MEx treated cultures. Conclusions: The present work establishes the protective effect of CP-MEx on IMR 32 Human Neuroblastoma cell line which is as much as by quercetin. The cytoprotective effect of CP-MEx was due to induction of antioxidant machinery of the cell hence holds therapeutic value in the treatment and/or prevention of neurodegenerative disorders of oxidative stress.