College of Animal Science and Veterinary, Jilin University, Changchun, Jilin, People's Republic of China.
Cell Mol Neurobiol. 2011 Apr;31(3):421-8. doi: 10.1007/s10571-010-9635-4. Epub 2010 Dec 18.
Depression is a major psychiatric disorder affecting nearly 21% of the world population and imposes a substantial health burden on society. Although significant progress has been made in depression research, the common molecular mechanism of antidepressants is still far from clearly understood. The neuroprotective effect of antidepressants has been proposed as a possible mechanism. Although Apocynum venetum (AV) L. (Apocynaceae) was previously shown to produce an antidepressant-like effect in the tail suspension test, the mechanisms underlying such antidepressant-like effect are yet to be understood. In this work, we studied the neuroprotective effect of AV leaf flavonoid extract in corticosterone-induced neurotoxicity, using PC12 cells as a suitable in vitro model of depression. Cell viability was quantitated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The release amount of lactic dehydrogenase (LDH) and intracellular Ca(2+) concentration were measured using kit, cell period change was tested by flow cytometry, and transcript abundances of brain-derived neurotrophic factor (BDNF) and microtubule-associated protein 4 (MAP4) were determined by real-time RT-PCR. The results showed that AV extract (25, 50, and 100 μg/ml) increased the A490 nm values, but decreased LDH release and Ca(2+) concentration, suppressed the apoptosis of PC12 cells and up-regulated BDNF and MAP4 transcript abundances compared with the corresponding corticosterone-treated group. These results suggest that the AV extract could generate a neuroprotective effect on corticosterone-induced neurotoxicity in PC12 cells, pointing to a possible action pathway by decreasing the Ca(2+) concentration and up-regulating BDNF and MAP4 genes.
抑郁症是一种主要的精神障碍，影响着全球近 21%的人口，给社会带来了巨大的健康负担。尽管在抑郁症研究方面取得了重大进展，但抗抑郁药的常见分子机制仍远未得到清晰理解。抗抑郁药的神经保护作用已被提出作为一种可能的机制。虽然先前的研究表明夹竹桃（Apocynum venetum (AV) L.）（夹竹桃科）在悬尾试验中产生抗抑郁样作用，但这种抗抑郁样作用的机制仍不清楚。在这项工作中，我们研究了夹竹桃叶黄酮提取物在皮质酮诱导的神经毒性中的神经保护作用，使用 PC12 细胞作为抑郁症的合适体外模型。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）测定法定量细胞活力。通过试剂盒测量乳酸脱氢酶（LDH）的释放量和细胞内 Ca(2+)浓度，通过流式细胞术测试细胞周期变化，通过实时 RT-PCR 测定脑源性神经营养因子（BDNF）和微管相关蛋白 4（MAP4）的转录丰度。结果表明，夹竹桃提取物（25、50 和 100 μg/ml）增加了 A490nm 值，但降低了 LDH 释放和 Ca(2+)浓度，抑制了 PC12 细胞的凋亡，并上调了 BDNF 和 MAP4 转录丰度与相应的皮质酮处理组相比。这些结果表明，夹竹桃提取物可对 PC12 细胞皮质酮诱导的神经毒性产生神经保护作用，提示通过降低 Ca(2+)浓度和上调 BDNF 和 MAP4 基因可能的作用途径。
