论文部分内容阅读
目的探讨不同浓度及不同时间的糖皮质激素诱导人气道上皮细胞9HTE0凋亡的机制。方法采用Annexin V/PI双染法检测低浓度组(0.3μmol/L)、中浓度组(3μmol/L)、高浓度组(10μmol/L)地塞米松(dexamethasone,Dex)作用6 h、12 h、24 h对人气道上皮细胞9HTE0早期凋亡的比例变化;RT-PCR法检测凋亡相关基因Bcl-2、Bax mRNA的表达情况;细胞免疫荧光法检测Bcl-2、Bax蛋白的定位及表达情况。结果 Annexin V/PI双染法检测中、高浓度组Dex在作用12 h、24 h后,早期凋亡率较正常组有显著性差异(P<0.05);Bcl-2 mRNA于高浓度组Dex作用6 h后即出现变化,而在中、高浓度组Dex作用12 h及24 h后,Bcl-2 mRNA均显著下降(P<0.05),Bax mRNA均显著增高(P<0.05);Bcl-2、Bax蛋白定位于细胞核和细胞浆中,中、高浓度组Dex作用24 h后荧光有明显变化。结论糖皮质激素作为哮喘一线用药,诱导了人气道上皮细胞9HTE0凋亡同时使其具有浓度和时间依赖性,从而为气道上皮的损伤提供了理论依据。
Objective To investigate the mechanism of glucocorticoid-induced apoptosis of human airway epithelial cells 9HTE0 at different concentrations and different times. Methods Annexin V / PI double staining was used to detect the effect of dexamethasone (Dex) in low concentration group (0.3μmol / L), middle concentration group (3μmol / L), high concentration group (10μmol / h, 24 h on the rate of early apoptosis of human airway epithelial cells 9HTE0; the expression of Bcl-2 and Bax mRNA was detected by RT-PCR; the localization of Bcl-2 and Bax protein was detected by immunofluorescence staining; Express the situation. Results Annexin V / PI double staining assay showed that there was a significant difference in the early apoptotic rate of Dex treated with Dex for 12 h and 24 h compared with the normal group (P <0.05). The expression of Bcl-2 mRNA in Dex (P <0.05), while Bax mRNA was significantly increased (P <0.05) after treated with dextran for 12 and 24 h in both medium and high concentration groups (P <0.05); Bcl- 2, Bax protein located in the nucleus and cytoplasm, medium and high concentrations of Dex group 24 h after significant changes in fluorescence. Conclusion Glucocorticoids, as a first-line agent for asthma, induces the apoptosis of human airway epithelial cells 9HTE0 in a concentration and time-dependent manner, which provides a theoretical basis for the injury of airway epithelium.