PLC-γ1信号通路在H_2O_2诱导的PC12细胞凋亡中的作用  

Role of phospholipase C-γ1 signaling pathway in H_2O_2-induced apoptosis of PC12 cells

作　　者：袁文丽[1,2] 陆地[3] 孙俊[3] 陈光学[1] 陈辉[1] 王廷华[1] 罗深秋[4] 

机构地区：[1]昆明医学院神经科学研究所,云南昆明650031 [2]云南省第二人民医院ICU,云南昆明650021 [3]昆明医学院人体解剖学教研室,云南昆明650031 [4]南方医科大学细胞生物学教研室,广东广州510515

出　　处：《南方医科大学学报》2008年第11期1939-1941,1946,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(30560170);昆明医学院中青年基金(03041)

摘　　要：目的探讨磷酸脂酶C-γ1(PLC-γ1)信号通路对H2O2诱导的PC12细胞凋亡作用的影响。方法利用PLC-γ1特异性抑制剂U73122(10μmol/L)预处理PC12细胞,阻断50μmol/LH2O2诱导的PLC-γ1信号通路,Hoechst/PI双染观察细胞形态改变,MTT评估细胞活力,流式细胞仪分析凋亡和坏死细胞比例,DNA电泳验证细胞凋亡状态的改变。结果H2O2对PC12细胞活力的影响呈时效和量效关系。PLC-γ1信号通路阻断后,PC12细胞出现了凋亡形态学改变,细胞活力明显降低,流式细胞仪检测出现典型的凋亡亚二倍体峰,凋亡细胞所占比例达35.7%,DNA电泳呈现明显的梯形条带。结论PLC-γ1信号通路在50μmol/LH2O2诱导的PC12细胞凋亡中发挥重要的保护作用。Objective To explore the role ofphospholipase C-γ1 （PLC-γ/1） signaling pathway in H2O2-induced apoptosis of PC12 cells. Methods PC12 cells were exposed to 50 μmol/L H2O2 after pretreatment with 10 μmol/L U73122, a specific PLC-γ/1 inhibitor. Hoechst/PI double staining was performed to observe the morphological changes of the cells under light microscope. MTT assay was used to evaluate the cell viability, and the percentage of apoptotic cells was analyzed by flow cytometry. DNA fi＇agmentation assay was carded out to characterize the cell apoptosis. Results After inhibition of the PLC-γ1 signaling pathway with 10 μmol/L U73122, PC12 Cells showed obvious apoptotic morphology, the viable cells decreased significantly, and the percentage of apoptotic cells rose to 35.7%. PC12 cells treated with U73122 presented with a distinct DNA ladder on electrophoresis resulting from DNA cleavage in the apoptotic cells. Conclusion PLC-71 signaling pathway plays an important protective role in HEO2-induced PC12 cell apoptosis.

关 键 词：磷酸脂酶C-γ1 U73122 H2O2 凋亡 

分 类 号：R651.2[医药卫生—外科学]