Hydroxysafflor Yellow A Protects PC12 Cells Against the Apoptosis Induced by Oxygen and Glucose Deprivation
Hydroxysafflor Yellow A Protects PC12 Cells Against the Apoptosis Induced by Oxygen and Glucose Deprivation
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DOI：
10.1007/s10571-011-9720-3
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发表时间：
2011-11-01
影响因子：
4
通讯作者：
Wang, Kunzheng
中科院分区：
文献类型：
--
作者：
Fan, Lihong;Dang, Xiaoqian;Wang, Kunzheng
羟基红花黄A （hydroxyysafflor yellow A， HSYA）在体内几种缺血性模型中具有神经保护作用。本研究探讨了HSYA对氧葡萄糖剥夺（OGD）诱导的急性神经元损伤的直接作用及其体外机制。采用缺氧葡萄糖剥夺（OGD） 4 h后再灌注（OGD - r） 20 h诱导分化的大鼠嗜铬细胞瘤PC12细胞体外缺血再灌注损伤。在缺血损伤前30分钟将HSYA（1、10和100 mu mol/l）添加到培养物中，并在OGD和再氧合阶段存在。MTT法检测PC12细胞存活率。生化法提高了丙二醛（MDA）含量和超氧化物歧化酶（SOD）活性。采用Hoechst 33258染色及流式细胞术检测细胞凋亡；western blotting检测Bcl-2、Bax、细胞色素C蛋白的表达。用比色法测定caspase-3活性。HSYA浓度依赖性地减轻神经元损伤，其特点是增加MTT损伤神经元的吸光度，减少细胞凋亡，拮抗OGD-R诱导的SOD活性降低和MDA水平升高。此外，HSYA以剂量依赖性的方式逆转Bcl-2的下调、Bax的上调以及线粒体细胞色素c向细胞质的释放以及由此引发的caspase-3的激活。这些结果表明，凋亡是ogd - r诱导的PC12死亡的一个重要特征，HSYA处理PC12细胞可以通过抑制细胞内氧化应激和线粒体依赖性caspase级联来阻断ogd - r诱导的凋亡。
Hydroxysafflor yellow A (HSYA) was reported neuroprotective under several ischemic models in vivo. In this study, the direct effect of HSYA against oxygen-glucose deprivation (OGD) inducing acute neuronal injury and the underling mechanisms in vitro were investigated. Four-hour oxygen and glucose deprivation (OGD) followed by 20 h reperfusion (adding back oxygen and glucose, OGD-R) was used to induce in vitro ischemia reperfusion injury in differentiated rat pheochromocytoma PC12 cells. HSYA (1, 10, and 100 mu mol/l) was added to the cultures 30 min prior to the ischemic insult and was present during OGD and reoxygenation phases. The survival rate of PC12 cells was detected by MTT assay. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) activity were elevated by biochemical method. Hoechst 33258 staining and flow cytometric analysis were used to detect apoptosis; western blotting was used to detect the expression of Bcl-2, Bax, and Cytochrome C protein. The activity of caspase-3 was assessed by colorimetry. HSYA concentration-dependently attenuated neuronal damage with characteristics of increasing injured neuronal absorbance of MTT, decreasing cell apoptosis, and antagonizing decreases in SOD activity and increase in MDA level induced by OGD-R. Moreover, the down-regulation of Bcl-2, up-regulation of Bax and the release of mitochondrial cytochrome c to cytosol and the consequent activation of caspase-3 were reversed by HSYA in a dose-dependent manner. These results suggest that apoptosis is an important characteristic of OGD-R-induced PC 12 death and that treatment of PC12 cells with HSYA can block OGD-R-induced apoptosis through suppression of intracellular oxidative stress and mitochondria dependent caspase cascade.