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目的:观察结核杆菌抗原(MTb-Ag)对BALB/c 3T3成纤维细胞水通道蛋白1(AQP1)表达的影响及其调节机制。方法:采用免疫印迹技术和免疫荧光检测细胞的AQP1和微管相关蛋白轻链3(LC3),实时定量PCR测定AQP1 mRNA含量,倒置显微镜观察细胞形态学变化。结果:低浓度MTb-Ag(10 mg/L)与细胞作用24~72 h后诱导细胞AQP1表达分别增加了74.46%、70.84%及93.89%(P<0.01),且细胞增长速度加快;而高浓度MTb-Ag(30 mg/L)并未诱导细胞AQP1表达增加。免疫荧光也证实低浓度的MTb-Ag诱导了细胞AQP1的显著增加。低浓度MTb-Ag(10 mg/L)作用于细胞24~72 h,均未检测出AQP1 mRNA的差异变化,但细胞的自噬反应显著增加。结论:MTb-Ag可诱导BALB/c 3T3成纤维细胞AQP1表达的显著增加,这种增加在于基因转录后的调节,而细胞的自噬途径可能参与其中。
Objective: To investigate the effect of MTb-Ag on the expression of aquaporin-1 (AQP1) in BALB / c 3T3 fibroblasts and its regulatory mechanism. Methods: AQP1 and LC3 were detected by immunoblotting and immunofluorescence. AQP1 mRNA was detected by real-time quantitative PCR. Morphological changes were observed by inverted microscope. RESULTS: AQP1 expression was increased by 74.46%, 70.84% and 93.89% (P <0.01) in cells induced by low concentration of MTb-Ag (10 mg / L) for 24-72 h and the cell growth accelerated The concentration of MTb-Ag (30 mg / L) did not induce the increase of cell AQP1 expression. Immunofluorescence also confirmed that low concentrations of MTb-Ag induced a significant increase in cellular AQP1. Low concentration of MTb-Ag (10 mg / L) in the cells 24 to 72 h, were not detected in AQP1 mRNA differences, but the cell autophagy increased significantly. CONCLUSION: MTb-Ag can induce a significant increase of AQP1 expression in BALB / c 3T3 fibroblasts. This increase is due to the regulation of gene transcription, and the autophagy pathway may be involved.