CAJ | 학술논문

目的：探讨神经酰胺对胶质瘤细胞87-MG和U251自噬性死亡的作用及机制。方法采用MTT和流式细胞的方法检测不同浓度神经酰胺刺激87-MG和U251细胞后细胞存活和凋亡的改变；电镜和Western blotting技术检测自噬和JNK/c-Jun信号通路的改变，JNK药理性抑制剂SP600125特异性抑制JNK通路，观察其对神经酰胺诱导自噬死亡的影响。结果神经酰胺刺激24 h后，87-MG和U251存活呈现时间依赖性下降(P<0.05)；相应的细胞死亡数目剂量依赖性升高(P<0.05)，但凋亡性死亡比例较低；神经酰胺刺激后镜下观察到的自噬小体计数，LC3B/LC3A和Beclin-1的表达以及JNK/c-Jun的磷酸化程度都增加(P<0.05)。提前给予SP600125抑制JNK信号通路的活性后，可以阻断神经酰胺诱导的细胞自噬性死亡(P<0.05)。结论神经酰胺可诱导胶质瘤细胞87-MG和U251出现自噬性死亡，机制可能与JNK信号通路的活化有关。
목적：탐토신경선알대효질류세포87-MG화U251자서성사망적작용급궤제。방법채용MTT화류식세포적방법검측불동농도신경선알자격87-MG화U251세포후세포존활화조망적개변；전경화Western blotting기술검측자서화JNK/c-Jun신호통로적개변，JNK약이성억제제SP600125특이성억제JNK통로，관찰기대신경선알유도자서사망적영향。결과신경선알자격24 h후，87-MG화U251존활정현시간의뢰성하강(P<0.05)；상응적세포사망수목제량의뢰성승고(P<0.05)，단조망성사망비례교저；신경선알자격후경하관찰도적자서소체계수，LC3B/LC3A화Beclin-1적표체이급JNK/c-Jun적린산화정도도증가(P<0.05)。제전급여SP600125억제JNK신호통로적활성후，가이조단신경선알유도적세포자서성사망(P<0.05)。결론신경선알가유도효질류세포87-MG화U251출현자서성사망，궤제가능여JNK신호통로적활화유관。
Objective To observe the autophagy of 87-MG and U251 glioma cells induced by ceramide and explore the possible mecha-nism. Methods The viability and apoptosis of 87-MG and U251 cells were detected by MTT assay and flow cytometry, respectively. Autoph-agic-related protein expressions of LC3B/LC3A and Beclin-1 were determined by Western blotting. The activation of JNK/c-Jun signaling pathway induced by ceramide with or without the treatment of JNK specific inhibitor SP600125 was also measured. Results 24 hours after treatment of ceramide, the growth of 87-MG and U251 cells was significantly inhibited time-dependently (P<0.05);and the number of au-tophagic cells increased dose-dependently (P<0.05). The levels of LC3B/LC3A and Beclin-1 significantly increased after ceramide treat-ment (P<0.05). JNK signaling pathway was activated in the 87-MG and U251 cells and the phosphorylation of c-Jun also increased after ce-ramide treatment. This activation of autophagy could be reversed by the pre-treatment of SP600125. Conclusion Ceramide may induce au-tophagy in 87-MG and U251 glioma cells and the mechanism may be related to the activation of JNK/c-Jun signaling pathway.