[金丝桃苷对高糖诱导心肌细胞氧化应激损伤的影响及机制]

[The Effect and Mechanism of Hyperoside on High Glucose-induced Oxidative Stress Injury of Myocardial Cells].

作者信息

Wang Cheng, Li Xia, Liu Zhen, Han Ming-Lei, Hou Yong-Lan, Guo Chang-Lei

机构信息

Deparment of Vasculocardiology, Xinxiang Central Hospital, Xinxiang 453100, China.

Department of Cardiology, First Affiliated Hospital of Xinxiang Medical College, Xinxiang 453100, China.

出版信息

Sichuan Da Xue Xue Bao Yi Xue Ban. 2018 Jul;49(4):518-523.

PMID:30378302

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 (<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 (<0.05).

CONCLUSION

Hyperoside protects myocardial cells against oxidative stress injury via activation of PI3K/AKT/Nrf2 signal pathway.

摘要

目的

探讨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。

方法

用高糖处理模拟心肌细胞氧化应激损伤。细胞分为五组：对照组（5.5 mmol/L葡萄糖）；高糖模型组（35 mmol/L葡萄糖）；金丝桃苷保护组（35 mmol/L葡萄糖+4、8、20 nmol/L金丝桃苷）。细胞孵育48小时。用CCK-8检测细胞活力。通过流式细胞术检测细胞凋亡。用活性氧检测试剂盒DCFH-DA通过流式细胞仪检测ROS水平。分别用SOD检测试剂盒和MDA检测试剂盒检测SOD和MDA水平。用蛋白质免疫印迹法检测磷脂酰肌醇3-激酶（PI3K）、蛋白激酶B（AKT）、p-AKT、核因子红细胞2相关因子2（Nrf2）和p-Nrf2的蛋白水平。通过免疫荧光染色分析AKT的激活情况。

结果

与对照组相比，高糖组细胞活力、SOD水平、PI3K表达、p-AKT/AKT和p-Nrf2/Nrf2比值以及AKT阳性细胞百分比降低，凋亡及ROS和MDA水平升高（<0.05）。与高糖组相比，金丝桃苷组（4、8、20 nmol/L）细胞活力、SOD水平、PI3K表达、p-AKT/AKT和p-Nrf2/Nrf2比值以及AKT阳性细胞百分比升高，凋亡及ROS和MDA水平降低（<0.05）。