绿原酸保护大鼠髓核细胞对抗氧化应激的作用机制研究

Mechanism of Chlorogenic Acid on Apoptosis of Rat Nucleus Pulposus Cells Induced by Oxidative Stress

目的:探讨绿原酸(CGA)对过氧化氢(H2O2)诱导的大鼠髓核细胞凋亡的作用机制。方法:采用体外培养大鼠髓核细胞,传代后取第3代细胞进行实验。随机分成5组:正常对照组、H2O2处理组、CGA+H2O2组、CGA组和CGA+PI3K/Akt抑制剂(LY294002)干预组。流式细胞术检测各组的细胞凋亡率及细胞内活性氧水平;Western blot检测各组髓核细胞中Akt,p-Akt,BCL-2的表达。结果:与正常对照组比较,H2O2组诱导的髓核细胞凋亡数显著增高,细胞内ROS含量显著增高;CGA预处理后显著抑制了髓核细胞凋亡和ROS产生,增加了细胞内p-Akt和BCL-2的表达,但LY294002抑制剂显著抑制了CGA对氧化应激后髓核细胞的上述功能。结论:绿原酸可通过激活PI3K-Akt信号途径抑制细胞内活性氧的产生、上调抗凋亡BCL-2蛋白水平的表达来保护髓核细胞免受氧化应激引起的损伤。

Objective：To investigate the mechanism of chlorogenic acid（CGA） on H2 O2-induced apoptosis in the rat nucleus pulposus ceils （NPCs）. Methods ： NPCs were isolated from SD rats and cultured in vitro. Cultured ceils （ P3 ） were randomly divided into normal control group, H2 O2 group, CGA ＋ H2 02 group, CGA group and LY294002 pretreatment group. The apoptosis and ROS production of rNPCs was detected by flow cytometry. The expressions of p-Akt, BCL-2 and Akt were analyzed by Western blot. Results ： Compared with normal control group, in the H2O2 group, the production of ROS and the apoptosis rate significantly increased in rNPCs;CGA treatment inhibited ROS production and cell apoptosis,while increased the expression of p-Akt and BCL-2;LY294002, a PI3Kinse inhibitor,not only decreased the expression of p-Akt and BCL-2, but also obviously increased ROS production and cell apoptosis. Conclusion ： Chlorogenie acid can protect NPCs against apoptosis by oxidative stress through decreasing reactive oxygen species production and increasing anti-apoptotic protein BCL-2 expression in NPCs by activation of PI3K-Akt signaling pathways.