Yuan Wen-Li, Lu Di, Sun Jun, Chen Guang-Xue, Chen Hui, Wang Ting-Hua, Luo Shen-Qiu
Institute of Neuroscience, Kunming Medical College, Kunming 650031, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2008 Nov;28(11):1939-41.
To explore the role of phospholipase C-gamma1 (PLC-gamma1) signaling pathway in H(2)O(2)-induced apoptosis of PC12 cells.
PC12 cells were exposed to 50 micromol/L H(2)O(2) after pretreatment with 10 micromol/L U73122, a specific PLC-gamma1 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 fragmentation assay was carried out to characterize the cell apoptosis.
After inhibition of the PLC-gamma1 signaling pathway with 10 micromol/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.
PLC-gamma1 signaling pathway plays an important protective role in H(2)O(2)-induced PC12 cell apoptosis.
探讨磷脂酶C-γ1（PLC-γ1）信号通路在过氧化氢（H₂O₂）诱导的PC12细胞凋亡中的作用。
用10微摩尔/升U73122（一种特异性PLC-γ1抑制剂）预处理PC12细胞后，将其暴露于50微摩尔/升H₂O₂中。进行Hoechst/PI双重染色，在光学显微镜下观察细胞的形态变化。采用MTT法评估细胞活力，通过流式细胞术分析凋亡细胞的百分比。进行DNA片段化分析以表征细胞凋亡。
用10微摩尔/升U73122抑制PLC-γ1信号通路后，PC12细胞呈现明显的凋亡形态，存活细胞显著减少，凋亡细胞百分比升至35.7%。经U73122处理的PC12细胞在电泳中呈现出由凋亡细胞中的DNA裂解产生的明显DNA梯带。
PLC-γ1信号通路在H₂O₂诱导的PC12细胞凋亡中起重要的保护作用。
