Background : Alzheimer`s disease (AD) is characterized by neuronal loss and extracellular senile plaque, whose major constituent is β-amyloid (Aβ), a 39-43 amino acid peptide derived from amyloid precursor protein. In cultures, Aβ can directly induce neuronal cell death and can render neurons vulnerable to excitotoxicity which may involve glutamate release and N-methyl-D-aspartate (NMDA) receptor. Silybum marianum (SM) has been used for centuries to treat liver disease due to its antioxidant, and anti-inflammatory properties. In particular, Silymarin, an active constituent of SM, has been reported to decrease lipid peroxidation. Therefore we hypothesized that SM might protect neurons against neurodegeneration in AD due to its antioxidant and anti-inflammatory activities. In the present study, the protective effect of ethanol extract from the stem of SM on Aβ (25-35)-induced neuronal cell death was examined in primary cultured rat cortical neurons. Methods and Results : Primary cultured cortical neurons were prepared using embryonic day 15 SD rat fetuses. Neurotoxicity experiments were performed on cultured neurons after 4-5 days in vitro. The cells were treated with 10 μM Aβ (25-35) or 1 mM NMDA for 36 h or 14 h, respectively. SM was applied 15 min before treatment of Aβ (25-35) or NMDA and also present in the medium during the incubations. The viability of neurons was monitored using a colorimetric MTT assay and Hoechst 33342 staining. The expression levels of anti-apoptotic and pro-apoptotic proteins were detected by western blot. An Ethanol extract of the stem of SM (10 and 50 μg/ml) significantly prevented Aβ (25-35)-induced apoptotic neuronal cell death in cultured cortical neurons. Furthermore SM inhibited Aβ (25-35)-induced decrease of anti-apoptotic protein, Bcl-2, and increase of pro-apoptotic proteins, Bax and active caspase-2, in western blot analysis. SM (10 and 50 μg/ml) also reduced NMDA-induced neuronal cell death. These results suggest that NMDA glutamate receptor activation is implicated in Aβ (25-35) -induced neuronal apoptotic death. Conclusion : The present study suggests that SM has a possible therapeutic role for preventing the progression of neurodegenerative disease such as Alzheimer's disease.

• Hae min Lee(College of Veterinary Medicine, Chungbuk National University) | 이해민
• Yeon Hee Seong(College of Veterinary Medicine, Chungbuk National University) | 성연희 Corresponding author