Neuroprotective Effect of Abelmoschus manihot Flower Extracts against the H(2)O(2)-Induced Cytotoxicity, Oxidative Stress and Inflammation in PC12 Cells

The progression of neurodegenerative diseases is associated with oxidative stress and inflammatory responses. Abelmoschus manihot L. flower (AMf) has been shown to possess excellent antioxidant and anti-inflammatory activities. This study investigated the protective effect of ethanolic extract (AME), water extract (AMW) and supercritical extract (AMS) of AMf on PC12 neuronal cells under hydrogen peroxide (H(2)O(2)) stimulation. This study also explored the molecular mechanism underlying the protective effect of AME, which was the best among the three extracts. The experimental results showed that even at a concentration of 500 μg/mL, neither AME nor AMW showed toxic effects on PC12 cells, while AMS caused about 10% cell death. AME has the most protective effect on apoptosis of PC12 cells stimulated with 0.5 mM H(2)O(2). This is evident by the finding when PC12 cells were treated with 500 μg/mL AME; the viability was restored from 58.7% to 80.6% in the Treatment mode (p < 0.001) and from 59.1% to 98.1% in the Prevention mode (p < 0.001). Under the stimulation of H(2)O(2), AME significantly up-regulated the expression of antioxidant enzymes, such as catalase, glutathione peroxidase and superoxide dismutase; promoted the production of the intracellular antioxidant; reduced glutathione; and reduced ROS generation in PC12 cells. When the acute inflammation was induced under the H(2)O(2) stimulation, AME significantly down-regulated the pro-inflammatory cytokines and mediators (e.g., TNF-α, IL-1β, IL-6, COX-2 and iNOS). AME pretreatment could also greatly promote the production of nucleotide excision repair (NER)-related proteins, which were down-regulated by H(2)O(2). This finding indicates that AME could repair DNA damage caused by oxidative stress. Results from this study demonstrate that AME has the potential to delay the onset and progression of oxidative stress-induced neurodegenerative diseases.