淫羊藿苷对H2O2诱导的软骨细胞氧化损伤的保护作用及其机制

Protective Effect of Icariin on H2O2-induced Chondrocyte Oxidative Damage and Its Mechanism

关键词:  淫羊藿苷  氧化损伤  凋亡  Nrf2/HO-1通路

Key words:  icariin  oxidative damage  apoptosis  Nrf2/HO-1 pathway

摘要:

目的 探究淫羊藿苷（icariin，ICA）对H₂O₂诱导的软骨细胞氧化损伤的保护作用及相关机制。方法 分离SD新生大鼠软骨细胞，随机分为对照组、H₂O₂模型组、ICA低剂量组、ICA中剂量组、ICA高剂量组；采用CCK8法检测各组细胞增殖能力的变化；采用ELISA试剂盒检测各组细胞中活性氧（reactive oxygen，ROS）、超氧化物歧化酶（superoxide dismutase，SOD）、丙二醛（malondialdehyde，MDA）、过氧化氢酶（catalase，CAT）以及谷胱甘肽过氧化物酶（glutathione peroxidase，GSH-Px）的表达情况；流式细胞术检测各组细胞周期情况，并计算增殖指数（proliferation index，PI）；Hoechst染色观察各组细胞核凋亡情况；分别采用荧光定量PCR （qRT-PCR）和Western blotting检测凋亡相关因子及Nrf2/HO-1通路的表达情况。结果 与对照组相比，H₂O₂模型组细胞增殖能力降低，ROS、MDA含量升高，SOD、CAT及GSH-Px含量下降，细胞凋亡情况加重；经ICA干预后，软骨细胞的增殖能力上升，ROS、MDA含量下降，SOD、CAT及GSH-Px含量增加，并且ICA能够有效抑制软骨细胞凋亡，上调Nrf2和HO-1蛋白的表达。结论 ICA对H₂O₂诱导的软骨细胞氧化损伤具有保护作用，能够抑制软骨细胞凋亡，其机制跟Nrf2/HO-1信号通路有关。

Abstract:

OBJECTIVE To explore the protective effect of icariin(ICA) on H₂O₂-induced chondrocyte oxidative damage and its related mechanism. METHODS SD neonatal rat chondrocytes were isolated and randomly divided into control group, H₂O₂ model group, ICA low dose group, ICA medium dose group and ICA high dose group. CCK8 assay was used to detect the change of cell proliferation ability in each group. The expression of reactive oxygen(ROS), superoxide dismutase(SOD), malondialdehyde(MDA), catalase(CAT) and glutathione peroxidase(GSH-Px) in each group were detected by ELISA kit. Cell cycle of each group was detected by flow cytometry and proliferation index(PI) was calculated. Hoechst staining was used to observe the cell apoptosis in each group. qRT-PCR and Western blotting were used to detect the expression of apoptosis related factors and Nrf2/HO-1 pathway. RESULTS Compared with the control group, the proliferation ability of cells in the H₂O₂ model group was decreased, ROS and MDA contents were increased, SOD, CAT and GSH-Px contents were decreased, and cell apoptosis was increased. After ICA intervention, the proliferation ability of chondrocytes increased, ROS and MDA contents decreased, SOD, CAT and GSH-Px contents increased, and ICA could effectively inhibit chondrocyte apoptosis and up-regulate the expression of Nrf2 and HO-1 proteins. CONCLUSION ICA has a protective effect on H₂O₂-induced oxidative damage of chondrocytes and can inhibit chondrocyte apoptosis, and the mechanism is related to Nrf2/HO-1 pathway.