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目的研究表没食子儿茶素没食子酸酯(EGCG)对神经细胞缺氧/复氧(A/R)损伤的影响及其作用机制。方法原代培养神经细胞,实验分4组:对照组、A/R组、EGCG预处理组(EGCG+A/R组)、EGCG+LY294002+A/R组。检测细胞存活率与乳酸脱氢酶(LDH)活性;流式细胞仪检测细胞凋亡和[Ca~(2+)]i;Western blot法检测总-Akt和磷酸化-Akt(p-Akt)蛋白表达。结果与对照组相比,A/R组中细胞存活率降低;LDH、[Ca~(2+)]i与细胞凋亡增加。与A/R组相比,EGCG预处理显著降低LDH、[Ca~(2+)]i与细胞凋亡,提高细胞存活率,表明EGCG可对抗神经细胞A/R损伤。进一步结果显示EGCG预处理后,显著增加神经细胞中p-Akt蛋白表达。然而,PI3K-Akt信号转导通路阻断剂LY294002显著抑制EGCG介导的神经保护作用与p-Akt蛋白上调。结论 EGCG对A/R介导的神经细胞损伤具有保护作用,其保护作用与PI3K-Akt信号转导通路有关。
Objective To study the effect of epigallocatechin gallate (EGCG) on the injury of nerve cells after hypoxia / reoxygenation (A / R) injury and its mechanism. Methods Primary cultured neurons were divided into 4 groups: control group, A / R group, EGCG pretreatment group (EGCG + A / R group), EGCG + LY294002 + A / R group. Cell viability and lactate dehydrogenase (LDH) activity were measured by flow cytometry. Apoptosis and [Ca ~ (2 +)] i were measured by flow cytometry. Total-Akt and phospho- Akt Protein. Results Compared with the control group, the cell viability decreased in A / R group; the apoptosis of LDH and [Ca ~ (2 +)] i cells increased. Compared with A / R group, EGCG pretreatment significantly reduced LDH, [Ca ~ (2 +)] i and apoptosis, and increased cell viability, indicating that EGCG could resist A / R injury of nerve cells. Further results showed that after EGCG pretreatment, the expression of p-Akt protein in nerve cells was significantly increased. However, PI3K-Akt signaling pathway blocker LY294002 significantly inhibited EGCG-mediated neuroprotection and up-regulation of p-Akt protein. Conclusion EGCG can protect A / R-induced neuronal injury and its protective effect is related to the PI3K-Akt signaling pathway.