二苯乙烯苷对大鼠原代皮层神经元氧糖剥夺损伤的保护作用

Protective effect of tetrahydroxystilbene glucoside on primary culture of rat cortical neurons against oxygen-glucose deprivation injury

目的：探讨二苯乙烯苷对大鼠皮层神经元氧糖剥夺（oxygen-glucose deprivation,OGD）损伤的作用。方法：取孕18 d的Sprague-Dawley胎鼠皮层神经元培养7-8 d,进行OGD损伤,同时分别加入二苯乙烯苷（10、20、40μg/m L）和尼莫地平（2.5μg/m L）进行处理。设置空白对照组、OGD组、OGD并加药组（二苯乙烯苷10、20、40μg/m L）以及OGD并尼莫地平阳性对照组。损伤6 h后,用噻唑蓝[3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide,MTT]法检测细胞活力,乳酸脱氢酶（lactate dehydrogenase,LDH）法检测LDH释放量,末端脱氧核苷酰基转移酶介导性d UTP切口末端标记（terminal deoxynucleotidyl transferase-mediated d UTP nick end-labeling,TUNEL）法检测细胞的凋亡,免疫蛋白印迹（Western Blot）法检测Toll样受体4（Toll-like receptor 4,TLR4）、核转录因子-κB（nuclear factor kappa B,NF-κB）及其抑制因子IκB和磷酸化的IκB（p-IκB）、白细胞介素-6（interleukin-6,IL-6）、p38及磷酸化的p38（p-p38）的蛋白表达。结果：OGD损伤后大鼠皮层神经元细胞活力显著降低,LDH释放量明显增多,细胞凋亡显著增多;二苯乙烯苷和尼莫地平均可显著抑制这一趋势。OGD损伤引起TLR4、NF-κB、p-IκB/IκB、p-p38/p38和IL-6的异常高表达,二苯乙烯苷明显抑制这些蛋白表达。结论：二苯乙烯苷对OGD诱导的原代皮层神经元损伤的保护作用可能是通过抑制TLR4-NF-κB炎症信号通路和p38通路发挥抗炎抗凋亡效应的。

Objective： To explore the effects and the possible mechanism of tetrahydroxystilbene glucoside on primary culture of rat cortical neurons against oxygen-glucose deprivation（OGD） injury. Methods： Cortical neurons of pregnant 18 d SpragueDawley fetal rats were in primary culture. MTT and LDH assay were used to examine cell viability. TUNEL assay was used to find cells apoptosis. Western Blot assay was used to study the possible mechanism. Results： Different concentrations of tetrahydroxystilbene glucoside significantly increased the cell viability,decreased LDH release and TUNEL positive apoptosis.Simultaneously,tetrahydroxystilbene glucoside could inhibit the upregulated protein level of TLR4,NF-κB,p-IκB/IκB,IL-6and p-p38/p38 induced by OGD injury. Conclusion： Tetrahydroxystilbene glucoside protected primary culture of rat cortical neurons against OGD injury. Simultaneously,the neuroprotective effects might be elicited by inhibiting the activation of TLR4-NF-κB and p38 signaling pathway to exhibit anti-inflammatory and antiapoptotic effects.