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Puerarin protects differentiated PC12 cells from H 2 O 2 ‐induced apoptosis through the PI3K/Akt signalling pathway
Author(s) -
Zhang Qin,
Huang Weidong,
Lv Xueying,
Yang Yunmei
Publication year - 2012
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20100900
Subject(s) - wortmannin , propidium iodide , apoptosis , pi3k/akt/mtor pathway , protein kinase b , puerarin , tunel assay , annexin , microbiology and biotechnology , programmed cell death , biology , terminal deoxynucleotidyl transferase , viability assay , chemistry , biochemistry , medicine , pathology , alternative medicine
Oxidative stress has been implicated as a major mechanism underlying the pathogenesis of neurodegenerative disorders. ROS (reactive oxygen species) can cause cell death via apoptosis. NGF (nerve growth factor) differentiated rat PC12 cells have been extensively used to study the differentiation and apoptosis of neurons. This study has investigated the protective effects of puerarin in H 2 O 2 ‐induced apoptosis of differentiated PC12 cells, and the possible molecular mechanisms involved. Differentiated PC12 cells were incubated with 700 μM H 2 O 2 in the absence or presence of different doses of puerarin (4, 8 and 16 μM). Apoptosis was assessed by MTS [3‐(4,5‐dimethylthiazol‐2‐yl)‐5‐(3‐carboxymethoxyphenyl)‐2‐(4‐sulfophenyl)‐2 H ‐tetrazolium] assay, TUNEL (terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labelling) analysis and Annexin V‐PI (propidium iodide) double staining flow cytometry. Protein levels of phospho‐Akt and phospho‐BAD (Bcl‐2/Bcl‐XL‐antagonist, causing cell death) were assayed by Western blotting. After stimulation with H 2 O 2 for 18 h, the viability of differentiated PC12 cells decreased significantly and a large number of cells underwent apoptosis. Differentiated PC12 cells were rescued from H 2 O 2 ‐induced apoptosis at different concentrations of puerarin in a dose‐dependent manner. This was through increased production of phospho‐Akt and phospho‐BAD, an effect that could be reversed by wortmannin, an inhibitor of PI3K (phosphoinositide 3‐kinase). The results suggest that puerarin may have neuroprotective effect through activation of the PI3K/Akt signalling pathway.