摘要
目的 探讨人参皂苷Rg1对MPP+ 诱导细胞凋亡保护作用的可能信号传导途径。方法 用吖啶橙 溴化乙锭染色观察SHSY5Y细胞凋亡率 ,流式细胞仪检测细胞内活性氧ROS水平 ,WesternBlotting法检测JNK(c junNH2 terminalkinase)激酶活性 ,免疫细胞化学染色法检测裂解的Caspase 3阳性细胞的表达率。结果 经 10 μmol·L- 1 Rg1或 2 5mmol·L- 1 N 乙酰半胱氨酸预处理后 ,MPP+ 诱导的SHSY5Y细胞凋亡受到明显抑制 ,同时细胞内ROS下降 ,JNK激酶的活性减弱 ,裂解的Caspase 3阳性细胞表达率下降。结论 Rg1可抑制MPP+ 诱导的SHSY5Y细胞凋亡 ,其作用机制可能是通过清除ROS、减弱JNK激酶的活性 ,从而减少Caspase 3的激活。
Aim To explore possible signal transmission way through which ginsenoside Rg1 protect cells from MPP + induced apoptosis. Methods The apoptosis of SHSY5Y induced by 1 methyl 4 phenylpyridinium (MPP +) was observed by AO EB staining. Flow cytometry was used to quantitate the reactive oxygen species (ROS). Western Blotting was used to detect the c jun NH 2 terminal kinase (JNK) activity in SHSY5Y cells. Immunocytochemistry staining was used to detect cleaved Caspase 3 positive cells. Results MPP + was shown to induce apoptosis in SHSY5Y cells. The percentage of apoptotic SHSY5Y cells induced by MPP + was obviously lower in those groups pretreated with 10 μmol·L -1 Rg1 or 2 5 mmol·L -1 N acetylcysyteine (NAC). It showed more ROS in MPP + groups than in control. JNK activity increased with time within 72 hours in 1 mmol·L -1 MPP + group. Simultaneously, it showed decrease of ROS, less activity of JNK and lower expression of cleaved Caspase 3 in 10 μmol·L -1 Rg1 and 2 5 mmol·L -1 NAC pretreated groups compared with groups treated with MPP + only. Conclusion Rg1 protects against MPP + induced apoptosis in SHSY5Y cells and the effect might be attributed to its removal of ROS, inhibition of the activity of JNK and expression of cleaved Caspase 3.
出处
《药学学报》
CAS
CSCD
北大核心
2003年第3期176-180,共5页
Acta Pharmaceutica Sinica
基金
福建省自然科学基金资助项目 (C0 110 0 15 )
