Wu Qian, Wang Ning, Wang Yan, Wang Guang-Yun, Piao Xin-Xin
Yao Xue Xue Bao. 2015 Feb;50(2):162-8.
To investigate the neuroprotective of ligustilide (LIG) against glutamate-induced apoptosis of PC12 cells, cell viability were examined by MTT assay. Flow cytometry was applied to assay cell apoptosis rate. Intracellular calcium concentration was measured by using fluorescent dye Fluo-3/AM. Cytochrome C (Cyt C), Caspase-3, Bax and Bcl-2 protein expression were assayed by western blot. The results showed that glutamate is cytotoxic with an inhibitory concentration 50 (ID50) of 15 mmol · L(-1). Pretreatment with LIG (1, 5, 15 μmol · L(-1)) significantly improved cell viability. The apoptosis rate in glutamate-induced PC12 cells was 13.39%, and decreased in the presence of LIG (1, 5, 15 μmol · L(-1)) by 9.06%, 6.48%, 3.82%, separately. Extracellular accumulation of Ca2+ induced by glutamate were significantly reduced by LIG. The results of western blot manifested that pretreatment LIG could decrease the release of Cyt C from mitochondria, down-regulate Caspase-3 protein expression and up-regulate Bcl-2/Bax ratio, thereby protects PC12 cells from apoptosis. In summary, LIG had protective effect on glutamate-induced apoptosis in PC12 cells through attenuating the increase in intracellular Ca2+ concentration, and inhibiting the release of Cyt C from mitochondria to cytoplasm.
为研究川芎嗪（LIG）对谷氨酸诱导的PC12细胞凋亡的神经保护作用，采用MTT法检测细胞活力。应用流式细胞术检测细胞凋亡率。使用荧光染料Fluo-3/AM测量细胞内钙浓度。通过蛋白质印迹法检测细胞色素C（Cyt C）、半胱天冬酶-3、Bax和Bcl-2蛋白表达。结果表明，谷氨酸具有细胞毒性，半数抑制浓度（ID50）为15 mmol·L⁻¹。用LIG（1、5、15 μmol·L⁻¹）预处理可显著提高细胞活力。谷氨酸诱导的PC12细胞凋亡率为13.39%，在LIG（1、5、15 μmol·L⁻¹）存在下分别降低9.06%、6.48%、3.82%。LIG可显著降低谷氨酸诱导的细胞外Ca²⁺积累。蛋白质印迹结果表明，LIG预处理可减少Cyt C从线粒体的释放，下调半胱天冬酶-3蛋白表达，上调Bcl-2/Bax比值，从而保护PC12细胞免于凋亡。综上所述，LIG通过减轻细胞内Ca²⁺浓度升高和抑制Cyt C从线粒体释放到细胞质，对谷氨酸诱导的PC12细胞凋亡具有保护作用。
