Cytoprotective propensity of Bacopa monniera against hydrogen peroxide induced oxidative damage in neuronal and lung epithelial cells

Abstract

Hydrogen peroxide (H 2 O 2 ), a major reactive oxygen species (ROS) produced during oxidative stress, is toxic to the cells. Hence, H 2 O 2 has been extensively used to study the effects of antioxidant and cytoprotective role of phytochemicals. In the present investigation H 2 O 2 was used to induce oxidative stress via ROS production within PC12 and L132 cells. Cytoprotective propensity of Bacopa monniera extract (BME) was confirmed by cell viability assays, ROS estimation, lipid peroxidation, mitochondria membrane potential assay, comet assay followed by gene expression studies of antioxidant enzymes in PC12 and L132 cells treated with H 2 O 2 for 24 h with or without BME pre-treatment. Our results elucidate that BME possesses radical scavenging activity by scavenging 2,2-diphenyl-1-picrylhydrazyl, 2,2 0 -azinobis(3-ethylbenzothiazoline-6-sulphonic acid), superoxide radical, and nitric oxide radicals. The IC 50 value of BME against these radicals was found to be 226.19, 15.17, 30.07, and 34.55 lg/ml, respectively). The IC 50 of BME against ROS, lipid peroxidation and protein carbonylation was found to be 1296.53, 753.22, and 589.04 lg/ml in brain and 1137.08, 1079.65, and 11101.25 lg/ml in lung tissues, respectively. Further cytoprotective potency of the BME ameliorated the mitochondrial and plasma membrane damage induced by H 2 O 2 as evidenced by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase leakage assays in both PC12 and L132 cells. H 2 O 2 induced cellular, nuclear and mitochondrial membrane damage was restored by BME pre-treatment. H 2 O 2 induced depleted antioxidant status was also replenished by BME pre-treatment. This was confirmed by spectrophotometric analysis, semi-quantitative RT-PCR and western blot studies. These results justify the traditional usage of BME based on its promising antioxidant and cytoprotective property.

Comparative evaluation of four triterpenoid glycoside saponins of bacoside A in alleviating sub-cellular oxidative stress of N2a neuroblastoma cells

“…To evaluate neuroprotective property of bacoside A components and mixture against stress in neuronal cells, H 2 O 2 was used as an oxidant to induce oxidative stress on N2a neuroblastoma cells. Previous studies have reported that H 2 O 2 is known to induce oxidative stress that cause lipid peroxidation, ROS generation, GSH depletion and reduction in mitochondrial membrane potential (MMP), antioxidant enzymes (SOD, catalase and GPx) activity . This oxidative stress was observed to be associated with increase in intracellular ROS .…”

“…superoxide dismutase, catalase and glutathione peroxidases in the neuronal cells . Bacosides‐rich extracts were evaluated for their anti‐apoptotic and cytoprotective properties on neuronal and lung epithelial cell lines . Existing studies had provided extensive documents on nootropic properties of B. monnieri but lack information on the role of four different bacoside A components in alleviating progression of neuronal diseases .…”

“…Moreover, increases in the intensity of Rh‐123 in different cells, post treatment with bacoside, were also observed indicating its ability to restore mitochondrial membrane potential. Role of bacoside A and its components on restoring mitochondrial membrane potential (MMP) is scantily documented . The study is even more important in the light of significance of bacoside A and its components and absence of any detailed analysis on the role of individual bacoside in regulating MMP.…”

“…Amongst four individual bacoside and their mixture, bacoside A3 and bacopaside II have shown around 2‐fold increased cell viability compare with stressed cells (Figure ). Previous studies have reported cytoprotective activity of bacosides rich extract against H 2 O 2 ‐induced cell mortality . Collectively, this study suggests that bacoside A components and bacoside mixture may be used as a neuroprotective compound with optimized dose having bacoside of specified concentration.…”

“…[16,17] Bacosides-rich extracts were evaluated for their anti-apoptotic and cytoprotective properties on neuronal and lung epithelial cell lines. [18,19] Existing studies had provided extensive documents on nootropic properties of B. monnieri but lack information on the role of four different bacoside A components in alleviating progression of neuronal diseases. [20] The lack of validation guidelines in most of the countries including developing nations like India also necessitates the need of analysing the role of bacoside A components for better efficacy towards neurological disorder.…”