白藜芦醇对新生大鼠神经元缺血缺氧时SDF-1/CXCR4通路抗凋亡作用的调控机制

Resveratrol modulates SDF-1/CXCR4 signaling to inhibit hypoxia-ischemia-induced apoptosis of neurons from the brain of newborn rats

目的:探究白藜芦醇(RSV)对缺血缺氧诱导的新生大鼠脑神经元凋亡及基质细胞衍生因子1(SDF-1)/CXC趋化因子受体4(CXCR4)通路的作用和机制。方法:体外培养新生大鼠皮质神经元,给予氧糖剥夺(OGD)模拟新生儿缺血缺氧性脑病(HIE)模型。首先将细胞随机分成4组:对照(control)组、HIE组、HIE+RSV低剂量(10μmol/L)组和HIE+RSV高剂量(50μmol/L)组。OGD处理2 h后加入相应剂量的RSV继续培养24 h,通过流式细胞术检测细胞凋亡,Western blot检测凋亡相关蛋白及SDF-1和CXCR4的蛋白表达水平,real-time PCR检测SDF-1和CXCR4的mRNA表达水平。然后使用CXCR4抑制剂AMD3100与RSV共同处理OGO损伤的细胞24h,检测SDF-1/CXCR4通路阻断后RSV对细胞凋亡和凋亡相关蛋白表达水平的影响。结果:RSV能够抑制OGD诱导的神经元凋亡,下调活化的caspase-3和细胞色素C的水平,上调Bcl-2/Bax比值(P<0.05)。与对照组相比,OGD导致SDF-1和CXCR4表达增加;与模型组相比,RSV能够上调SDF-1和CXCR4的表达(P<0.05);使用AMD3100阻断SDF-1/CXCR4通路减弱了RSV抑制OGD诱导的细胞凋亡的作用。结论:RSV可抑制缺血缺氧诱导的新生大鼠神经元凋亡,其机制可能是通过上调SDF-1/CXCR4通路发挥作用的。

AIM： To investigate the effect of resveratrol（ RSV） on apoptosis and stromal cell-derived factor-1（ SDF-1）/CXC chemokine receptor 4（ CXCR4） pathway in hypoxia-ischemia-induced neurons from the brain of newborn rats and its mechanism. METHODS： The cortex neurons from the brain of newborn rats were given oxygen-glucose deprivation（ OGD） treatment to mimic neonatal hypoxic-ischemic encephalopathy（ HIE）. The cortex neurons were randomly divided into 4 groups： control group,HIE model group,HIE ＋ RSV-low（ 10 μmol/L） group,and HIE ＋ RSV-high（ 50μmol/L） group. After OGD treatment for 2 h,the neurons were cultured with indicated dose of RSV for 24 h. The apoptosis was analyzed by flow cytometry. Western blot was used to determine the levels of apoptosis-related proteins,SDF-1 and CXCR4. Real-time PCR was used to detect the mRNA expression of SDF-1 and CXCR4. Additionally,to explore the effects of RSV on cell apoptosis and apoptosis-related proteins after the suppression of SDF-1/CXCR4 signaling,a CXCR4 antagonist AMD3100,and RSV were used to co-treat OGD-injured neurons for 24 h. RESULTS： RSV alleviated OGD-induced neuronal apoptosis,down-regulated cleaved caspase-3 and cytochrome C levels,and up-regulated the ratio of Bcl-2/Bax. Compared with the control group,OGD treatment increased the expression of SDF-1 and CXCR4（ P〈0. 05）. Compared with the HIE model group,RSV further up-regulated the expression of SDF-1 and CXCR4（ P〈0. 05）. AMD3100 reversed the effects of RSV on OGD-induced cell apoptosis. CONCLUSION： RSV suppresses hypoxia-ischemia-induced apoptosis of neurons from the brain of newborn rats via up-regulating SDF-1/CXCR4 signaling pathway.