Neuropharmacology and AnalgesiaPotassium 2-(1-hydroxypentyl)-benzoate attenuated hydrogen peroxide-induced apoptosis in neuroblastoma SK-N-SH cells
Introduction
Oxidative stress is one of the mechanisms involved in neuronal damage. A number of evidence indicate that oxidative stress plays a crucial role in cerebral ischemic stroke as well as neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, Amyotropic lateral sclerosis and so on (Mariani et al., 2005, Nunomura et al., 2001, Ozkul et al., 2007). In the normal brain tissue, the production of reactive oxygen species and reactive nitrogen species is balanced by endogenous enzymatic and nonenzymatic antioxidative defenses. After cerebral ischemia and particularly reperfusion, the free radical production is dramatically increased and overwhelms endogenous antioxidant systems, leading to a disruption of the equilibrium and neuronal death (Lipton, 1999, Reddy et al., 2009). In stroke patients, the lipid peroxides, such as malondialdehyde, lipid peroxides, are increased in the plasma and erythrocytes (Alexandrova et al., 2004, Sharpe et al., 1994). In Alzheimer's disease patients, the study showed that oxidative stress induced DNA damage (Nunomura et al., 2001), lipid peroxidation (Perry et al., 2002) and neuronal death (Chandra et al., 2000). Thus, the therapeutic strategies of inhibiting oxidative stress-induced neuronal damage appear to be promising in the treatment of stroke and Alzheimer's disease.
Previous studies have demonstrated that oxidative stress-induced neuronal death was partially attributable to apoptosis. In the acute stroke, the neurons in the penumbra which are near to the infarct area, might develop to apoptotic death or recover with the anti-oxidative treatment. Studies on the postmortem tissues of Alzheimer's disease patients provided the direct morphological and biochemical evidence that some neurons in the brain degenerated via an apoptotic mechanism that included the presence of DNA damage, nuclear apoptotic bodies, and other markers of apoptosis (Chen et al., 2001, Janicki and Monteiro, 1997). Thus, blocking apoptosis might be important for prevention of neuronal death during stroke and the neurodegenerative diseases.
Potassium 2-(1-hydroxypentyl)-benzoate is a novel drug candidate for treatment of cerebral ischemia. Previous studies showed that dl-PHPB reduced infarct volume and improved the neurobehavioral deficits in the cerebral ischemic rat model (Zhang et al., 2006). Furthermore, dl-PHPB was shown to improve the learning and memory deficits in the cerebral hypoperfused rats, Aβ25–35 intracerebroventricular-infused rats and the senescence-accelerated mice (SAM) (unpublished). In 2009, dl-PHPB was approved by the State Food and Drug Administration of China (SFDA) as an anti-stroke and neuroprotective drug for phase I clinical trial. Though, dl-PHPB was shown to increase cerebral blood flow and inhibit platelet aggregation, the effect of dl-PHPB on oxidative damage of neurons remains to be elucidated. In the present study, we investigated the neuroprotective effect of dl-PHPB on H2O2-induced apoptosis and the related mechanisms in the neuroblastoma SK-N-SH cells.
Section snippets
Materials
Dl-PHPB was provided by the Department of Synthetic Pharmaceutical Chemistry, Institute of Materia Medica with a purity of 98.5%. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Hoechst33342 and β-actin antibody were purchased from Sigma (St. Louis, MO, USA). PKCα antibody was purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Bax, Bcl-2 and Caspase3 were purchased from Santa Cruz biotechnology (Santa Cruz, CA, USA).
Cell culture and experimental treatment
SK-N-SH human neuroblastoma
Dl-PHPB attenuated H2O2-induced cell death
In order to choose the optimal H2O2 concentration, we performed the dose–response curves of H2O2-induced cell death firstly. The concentration of H2O2 was set from 50 μM to 500 μM. The result showed that H2O2 at concentration of 150 μM induced adequate cell death, thus the concentration was chosen in the further study (Fig. 1A). Fig. 1B showed the percentage of the cell viability after 24 h exposure to 150 μM H2O2 in the presence of different concentrations of dl-PHPB. The result showed that 150 μM H2
Discussion
Neurodegenerative diseases and stroke are the leading cause of adult disability in the developed countries. Although they have different risk factors and pathophysiological mechanisms, oxidative stress and neuronal apoptosis are considered to be involved in both conditions, leading to cell damage (Yu et al., 2008). The productions of free radicals and other chemical species have been demonstrated to increase in both stroke and neurodegenerative disease (Butterfield et al., 2001, Cuzzocrea et
Acknowledgements
The study was supported by National Major Special Project on New Drug Innovation of China (2012ZX09301002-004) and National 973 Project (2011CB504103).
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Present address: College of Pharmaceutical Sciences, Shihezi University, Shihezi 832002, China.