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金丝桃苷对高糖诱导的心肌细胞氧化应激损伤的影响及其机制研究
The Effect and Mechanism of Hyperoside on High Glucose-induced Oxidative Stress Injury of Myocardial Cells
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摘要: 目的 探索金丝桃苷在高糖诱导的心肌细胞氧化应激损伤中的作用及其分子机制。 方法 高糖处理模拟心肌细胞氧化应激损伤。细胞分为5个组：正常对照组（5.5 mmol/L葡萄糖），高糖损伤模型组（35 mmol/L葡萄糖），低、中、高浓度金丝桃苷保护组（35 mmol/L葡萄糖+4/8/20 nmol/L金丝桃苷）。各组细胞培养48 h后，CCK-8检测细胞存活力；流式细胞术分析细胞凋亡；通过流式细胞仪利用活性氧（ROS）检测试剂盒DCFH-DA分析ROS水平；超氧化物歧化酶（SOD）和丙二醛（MDA）试剂盒检测SOD和MDA水平；Western blot法检测磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B（AKT）、磷酸化（p）-AKT、核因子E2相关因子2（Nrf2）、p-Nrf2的表达；免疫荧光染色分析AKT的活化情况。 结果 与正常对照组比较，高糖损伤模型组细胞存活率降低，细胞凋亡率增高，ROS、MDA水平升高，SOD水平降低，PI3K相对表达量及AKT、Nrf2磷酸化水平（p-AKT/AKT和p-Nrf2/Nrf2的比值）降低，AKT阳性细胞数比率降低，以上差异均有统计学意义（P<0.05）。与高糖损伤模型组相比，金丝桃苷保护组（4、8、20 nmol/L）细胞存活率升高，细胞凋亡率降低，ROS、MDA水平降低，SOD水平升高，PI3K相对表达量及AKT、Nrf2磷酸化水平升高，AKT阳性细胞数比率升高，以上差异均有统计学意义（P<0.05）。 结论 金丝桃苷可通过激活PI3K/AKT/Nrf2信号通路保护心肌细胞免受高糖诱导的氧化应激损伤。Abstract: Objective To explore the effect and mechanism of hyperoside on high glucose-induced oxidative stress injury of myocardial cells. Methods Oxidative stress injury of myocardial cells was simulated by treating with high glucose. Cells were divided into five groups:control group (5.5 mmol/L glucose); high glucose model group (35 mmol/L glucose); hyperoside protection groups (35 mmol/L glucose +4, 8, 20 nmol/L hyperoside). Cells were incubated for 48 h. The cell viability was detected by CCK-8. Apoptosis was measured through flow cytometry. The level of ROS was tested by Reactive Oxygen Species Assay Kit DCFH-DA with flow cytometer. The level of SOD and MDA was detected by SOD Assay Kit and MDA Assay Kit respectively. The protein levels of phosphatidylinositide 3-kinases (PI3K), protein kinase B (AKT), p-AKT, nuclear factor erythroid 2-related factor 2 (Nrf2) and p-Nrf2 were detected by Western blot. The activation of AKT was analyzed by immunofluorescence staining. Results Compared with control group, the cell viability, the levels of SOD, the expression of PI3K, the ratio of p-AKT/AKT and p-Nrf2/Nrf2 and the percentage of AKT positive cells in high glucose group were decreased with enhancive apoptosis and levels of ROS and MDA (P<0.05). Compared with high glucose group, the cell viability, the levels of SOD, the expression of PI3K, the ratio of p-AKT/AKT and p-Nrf2/Nrf2 and the percentage of AKT positive cells in hyperoside group (4, 8, 20 nmol/L) were increased with reduced apoptosis and levels of ROS and MDA (P<0.05). Conclusion Hyperoside protects myocardial cells against oxidative stress injury via activation of PI3K/AKT/Nrf2 signal pathway.
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Keywords:
- Hyperoside /
- Oxidative stress injury /
- Apoptosis /
- PI3K /
- AKT /
- Nrf2
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