摘要
目的研究2,3,5,4'-四羟基二苯乙烯-2-O-β-D-葡萄糖苷(2,3,5,4'-tetrahydroxystibene-2-O-β-D-glucoside,TSG)对1-甲基-4-苯基吡啶离子(1-methy-4-phenylpyridinium;MPP+)诱导PC12细胞凋亡的影响及其可能机制。方法 4-甲基偶氮唑蓝(MTT)检测PC12细胞活性;Hochest 33258染色观察细胞凋亡的变化;DCFH-DA/DAF-FM DA检测ROS/NO表达情况;Westernblotting检测Nrf2、Keap1、SOD1、SOD2、CAT蛋白的表达情况。结果 MPP+(500μmol·L^(-1))作用于PC12细胞24 h后,与正常组比较,细胞存活率降低至(51.3±2.2)%(P<0.01);TSG(10,50μmol·L^(-1))预处理24 h后,细胞存活率增加至(60.1±1.5)%,(74.2±2.1)%,(82.1±1.5)%(P<0.05);细胞核固缩明显减少,且具有剂量依赖性关系。此外,MPP+处理后使PC12细胞中ROS/NO升高,Nrf2、SOD1、SOD2、CAT蛋白表达减少,使keap1表达增多,而TSG预处理可以明显改善这些变化。结论 TSG对MPP+诱导的PC12细胞凋亡具有浓度依赖性的抑制作用,其作用机制可能通过激活Nrf2/Keap1通路减轻细胞内活性氧的聚集有关。
OBJECTIVE To explore the protective effect of 2,3,5,4＇-tetrahydroxystibene-2-O-β-D-glucoside （TSG） on 1-methy- 4-phenylpyridinium（ MPP ＋ ） induced apoptosis in PC12 cells and its possible mechanism. METHODS The cell viability was meas- ured by 3- [ 4,5-dimethyhhiazol-2-yl] -2,5-diphenyltetrazolium bromide （ MTT）. The morphological change of PC 12 ceils was observed by Hoechst 33258 staining. The intracellular ROS/NO level was examined by using DCFH-DA/DAF-FM DA. The expression level of Nrf2, Keapl, SOD1, SOD2, CAT protein was detected by Western blotting. RESULTS Compared with the control group, the ratio of cell survival decreased to （51.3±2. 2） % （P 〈0. 01 ） after MPP＋ treatment with PCl2 cells for 24 h. Cell viability was increased to （60.1± 1.5）%, （74.2 ± 2.1）%, （82.1 ± 1.5）% （P 〈 0.05） respectively after exposure with 10, 50 μmol·L-l TSG. Cytochromatin concentration reduced and show the relation between quantity and result. Furthermore, TSG inhibited the MPP ＋ - induced increase reactive oxygen species/nitric oxide （ROS/NO） in PCl2 cell, and counteracted the over expression of Keapl and low-expression of Nrf2, SOD1 , SOD2, CAT. CONCLUSION TSG plays a possible neuroprotective role in MPP ＋ induced apoptosis of PCl2 cells by a concentration-dependent manner, and the mechanism of the effects of TSG may be involved in facilitating Nrf2/keapl activation.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2017年第11期948-953,共6页
Chinese Pharmaceutical Journal
基金
国家自然科学基金资助项目(30772743)
