羟基红花黄色素A抗缺氧/复氧所致神经元损伤的作用及机制研究

Effect of hydroxysafflor yellow A(HSYA) on neuron against hypoxia/reoxygenation injury and its mechanism

关键词:  羟基红花黄色素A  神经元  缺氧/复氧  突触可塑性

Key words:  hydroxysafflor yellow A  neuron  hypoxia/reoxygenation  synaptic plasticity

摘要:

[目的] 考察羟基红花黄色素A（HSYA）抗缺氧/复氧（H/R）损伤神经元的作用及机制。[方法] 原代培养Wistar大鼠胎鼠皮层神经元，将神经元随机分为正常对照组、H/R组、羟基红花黄色素A低（5 μg/mL）、中（20 μg/mL）、高（80 μg/mL）剂量组，考察HSYA对神经元活力和乳酸脱氢酶（LDH）释放的影响；采用逆转录-聚合酶链式反应（RT-PCR）和酶联免疫吸附实验（ELISA）分别检测脑源性神经生长因子（BDNF）、生长相关蛋白43（GAP-43）及Nogo-A等与神经元突触重塑相关指标的mRNA和蛋白表达。[结果] 与正常对照组比较，H/R组细胞活力明显降低，LDH释放明显增加（P<0.01），BDNF及GAP-43 mRNA和蛋白表达均显著降低（P<0.05），Nogo-A mRNA和蛋白表达显著增加（P<0.01）；与H/R组比较，HSYA低、中、高3个剂量组细胞活力均显著增强，LDH释放明显减少（P<0.01），BDNF及GAP-43 mRNA及蛋白表达明显增加，Nogo-A mRNA和蛋白表达显著降低（P<0.05或P<0.01）。[结论] HSYA能够提高神经元活力，减少LDH释放，具有抗H/R所致神经元损伤的作用，该作用与促进BDNF表达有关；此外，HSYA还能促进GAP-43 mRNA和蛋白表达、抑制Nogo-A mRNA和蛋白表达，从而增加神经元突触可塑性。

Abstract:

[Objective] To introduce HSYA might act as a potential neuroprotective agent against the hypoxia/reoxygenation(H/R)injury in neuron. [Methods] Cortical neurons of Wistar rat were primary cultured in vitro. The cortical neurons were divided randomly into five groups:control, H/R and HSYA(5, 20, 80 μg/mL)+H/R. By observing the changes of cell viability and the releasing of LDH, proved the effect of HSYA on neuron injuried by H/R. The expressions of BDNF, GAP-43 and NogoA were studied. By using real-time RT-PCR and ELISA. [Results] Compared with control group, the cell viability decreased and the releasing of LDH increased significantly(P<0.01), the mRNA and protein expressions of BDNF and GAP-43 decreased significantly(P<0.05), the mRNA and protein expression of NogoA increased significantly(P<0.01) in H/R group. Compared with control group, three dosages of HSYA increased cell viability and reduced the releasing of LDH significantly(P<0.01), and increased the mRNA and protein expressions of BDNF and GAP-43, and decreased the mRNA and protein expressions of NogoA(P<0.05 or P<0.01). [Conclusion] HSYA could increase cell viability, and reduce the releasing of LDH, thus relieving the neuronal damage caused by H/R, which related to promoting BDNF expression. In addition, HSYA can also promote the neuronal synaptic plasticity by promoting the expression of GAP-43 mRNA and protein, and inhibiting the expression of Nogo-An mRNA and protein.