白藜芦醇通过调节 SIRT1/AMPK 信号通路保护氧-葡萄糖剥夺大鼠皮质神经元
Resveratrol protects oxygen-glucose deprivation rat cortical neurons by regulating SIRT1/AMPK signaling pathways
摘要目的:探讨白藜芦醇对单次和双次氧-葡萄糖剥夺(oxygen-glucose deprivation, OGD)原代皮质神经元沉默信息调节因子1(silent information regulator 1, SIRT1)、AMP 活化的蛋白激酶(AMP-activated protein kinase, AMPK)活性和 ATP 含量的影响及其可能的神经保护机制。方法皮质神经元取自18 d Wistar 大鼠胚胎,原代培养成功后通过2次OGD 制作体外反复缺血模型。采用台盼蓝染色法检测细胞存活率,蛋白质印迹法检测 SIRT1和磷酸化 AMPK 表达,去乙酰化酶荧光检测法检测 SIRT1活性,生物发光测定法检测 ATP 含量。结果与对照组比较,白藜芦醇(0.5μmol/L)预处理能显著增高单次和双次 OGD 后存活率(P 均<0.001)、ATP 含量(P 均=0.004)、SIRT1活性(单次:P =0.001;双次:P =0.002)以及 SIRT1(单次:P =0.029;双次:P =0.023)和磷酸化 AMPK (P 均=0.001)表达水平。结论白藜芦醇对单次和双次 OGD 皮质神经元均具有神经保护作用,其机制可能与上调 SIRT1/AMPK 通路和降低能量需求有关。
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abstractsObjective To investigate the effects of resveratrol on the first and double oxygen-glucose deprivation (OGD) primary cortical neuron silent information regulator 1 (SIRT1), AMP-activated protein kinase (AMPK) activity and ATP content, and its possible neuroprotective mechanism. Methods Cortical neurons were taken from the embryos of 18-day Wistar rats. An in vitro repeated ischemia model was induced by the double OGD after the success of primary culture. Trypan blue stalning was used to detect the cel survival rate. Western blot was used to detect the SIRT1 and phospho-AMPK expression. Deacetylase fluorescence assay was used to detect the SIRT1 activity. Bioluminescence assay was used to detect the ATP content. Results Compared with the control group, resveratrol (0. 5 μmol/L) preconditioning significantly increased the survival rates after the single and double OGD (al P < 0. 001), ATP content (al P = 0. 004), SIRT1 activity (single: P = 0. 001; double: P = 0. 002), and the expression levels of SIRT1 (single: P = 0. 029; double: P = 0. 023) and phospho-AMPK (al P = 0. 001). Conclusions Resveratrol has the neuroprotective effect for the first and double OGD cortical neurons. Its mechanism may be associated with upregulating the SIRT1/AMPK signaling pathways and decreasing the energy requirements.
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