摘要
目的探讨MEK1信号通路与支气管上皮细胞(16-HBE)谷氨酰半胱氨酸合成酶(1-GCS)表达的关系。方法应用MEK1抑制剂PD9805950μmol/L分别孵育16-HBE2、8、16及24h。用Western印迹方法和荧光定量PCR检测细胞γ-GCS重链蛋白和mRNA的表达。用显色方法检测细胞内谷胱甘肽(GSH)的含量。用Western印迹方法检测细胞c-jun的水平。结果经PD98059孵育4、8、16及24h后,γ-GCS重链蛋白和mRNA的表达和细胞内GSH的含量较对照组均增加。各时间段c-jun表达较各对照组减少。结论在支气管上皮细胞中MEKI抑制剂PD98059对16-HBE的γ-GCS和GSH有明显的上调作用。阻断激活蛋白-1(AP-1)途径不能减少γ-GCS和GSH的合成。
Objective To explore the relation between the MEKI signal transduction pathway and the synthesis of-y-glutamylcysteine synthetase(γ-GCS) in the bronchial epithelial cell( 16-HBE). Methods 16-HBE cells were incubated with the PD98059(50 μmol/L) for 2,8,16 and 24 hours. The expression of γ-GCS mRNA and protein was measured by Western blotting and real-time PCR. Cellular GSH level was detected by staining method. C-jun was examined by Western blotting. Results After the incubation with the PD98059 for 4,8,16 and 24 hours, the expression of γ-GCS, γ-GCS mRNA and GSH increased, but the expression of c-jun decreased as compared to the control group. Conclusions PD98059, inhibitor of MEKI, can increase the synthesis of γ-GCS and GSH in the 16-HBE. Blocking AP-1 could not decrease the synthesis of γ-GCS and GSH.
出处
《中华生物医学工程杂志》
CAS
2007年第4期195-197,共3页
Chinese Journal of Biomedical Engineering
基金
广东省自然科学基金团队(05200239)