✍ Scribed by Kyu-Yong Lee; Seong-Ho Koh; Sangjae Kim; Hyun-Hee Park; Young Joo Lee
No coin nor oath required. For personal study only.
We investigated the effect of hetastarch, used for the treatment of acute ischemic stroke, on neuronal cell damage by oxidative stress, a main pathogenic mechanism in ischemic stroke. Neuronally differentiated PC12 cells (nPC12 cells) were treated with varying concentrations of hetastarch and hydrogen peroxide (H~2~O~2~), and their viability was measured with a 3,(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT) assay and trypan blue staining. The effect of hetastarch on free radical production by H~2~O~2~ was evaluated using the fluorescent probe 2′,′‐dichlorodihydrofluorescein diacetate (DCFH‐DA) and by quantifying the amount of 2,5‐ and 2,3‐dihydroxybenzoic acid (DHBA). Additionally, the expression levels of BAX, Bid, Bcl‐2, Bcl‐xL, cytosolic cytochrome c, and cleaved caspase‐3 were examined using Western blot analysis. Following exposure to 100 µM H~2~O~2~, the viability of nPC12 cells significantly decreased; however, cell viability increased with hetastarch treatment. Free radical production related to H~2~O~2~ exposure was significantly reduced after 100 µM hetastarch treatment. The expression levels of BAX, Bid, cytosolic cytochrome c, and activated caspase‐3 were reduced, whereas Bcl‐2 and Bcl‐xL levels increased in H~2~O~2~‐injured nPC12 cells treated with 100 µM hetastarch, as compared with nPC12 cells that were treated with only 100 µM H~2~O~2~. These results demonstrate that hetastarch can reduce oxidative stress‐induced neuronal cell death. Drug Dev Res 73: 35–42, 2012. © 2011 Wiley Periodicals, Inc.
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