不同剂量米非司酮诱导Nrf2基因表达对百草枯致A549细胞损伤的影响

The effects of Nrf2 gene expression induced by RU486 at different doses on A549 cell damage induced by paraquat

目的观察不同剂量米非司酮（RU486）诱导Nrt2基因表达对百草枯（PQ）致A549细胞毒性影响。方法Ad—RUNrf2感染的A549细胞，在给予10^-10、10^-9、10^-8、10^-7mol／L的RU486诱导表达6h后，加入浓度为10^-3mol／L的PQ培养48h，用实时定量聚合酶链式反应（real．timePCR）法和凝胶迁移率试验（EMSA）检测不同浓度RU486诱导的Nrf2表达情况；Real-timePCR、ELISA检测不同浓度Nrf2对PO致A549细胞损伤炎症因子白细胞介素（IL）-6、IL-10、肿瘤坏死因子（TNF）-α，凋亡因子半胱氨酸天冬氨酸蛋白酶（Caspase）-3、Caspase-9、细胞色素c（CytC）基因相对表达量和蛋白含量的影响。用化学比色法检测氧化因子过氧化氢酶（CAT）、丙二醛（MDA）蛋白含量。结果Nrf2基因相对表达量及蛋白表达均随RU486浓度增高而增高，其中以RU486浓度为10^-7mol／L时最高，与PQ中毒组及空白对照组的差异有统计学意义（P〈0．01，P〈0．05）。随RU486浓度降低，IL．6、TNF一仪的基因相对表达量及蛋白含量升高，在RU48610。mol／L时最低，与PO中毒组及空白对照组的差异有统计学意义（P〈0．01，P〈0．05）；IL-10的变化则相反。随RU486浓度降低，Caspase-3、Caspase-9、CytC基因相对表达量升高，10^-7mol／L时最低，与PQ中毒组及空白对照组的差异有统计学意义（P〈0．01，P〈0．05）。随着RU486浓度的增高，CAT蛋白含量增高，RU486浓度为10^-7mol／L时含量最高，与PQ中毒组及空白对照组的差异有统计学意义（P〈0．05），MDA含量的变化则相反。结论RU486诱导Nrf2基因表达能促进A549细胞的氧化一抗氧化系统平衡，并抑制炎症因子、凋亡因子，对PO致A549细胞损伤有保护作用。

Objective To observe the effects of Nrf2 gene expression induced by RU486 at different doses on A549 cell damage induced by paraquat （PQ）. Methods After A549 cells transfected with Ad- RUNrf2 were treated by RU486 at the doses of 10-10, 10-9, 10-6 and 10-7 mol / L for 6 h, A549 cell cultures were exposed to 10-3 mol/L of PQ for 48 h. Then qRT-PCR and EMSA assays were used to detect the expression of Nrf2 gone, and qRT-PCR and ELISA assays were utilized to measure the effects of Nrf2 gene on the expression of the inflammatory cytokines IL-6, IL-10 and TNF-α, apoptotic factors Caspase-3, Caspase-9 and Cytochrome C. The oxidation factors （CAT and MDA protein contents） were observed by Chemical Calorimetric Analysis. Results NT2 gene relative expression and protein contents increased with RU486 concentrations, and the above expression was the highest when the concentration of RU486 was 10 mol/L, which was significantly higher than those in control and PQ exposure groups （P〈0.01 or P〈0.05）. The relative gene expression and protein expression of IL-6 and TNF-α enhanced with the reduced concentrations of RU486, which were the lowest when RU486 concentration was 10-7 mol/L, as compared with control and PQ exposure groups （P〈0.01 or P〈0.05 ）, while the change of IL-10 content was the opposite. The relative expression of Caspase3, Caspase9 and Cytochrome C genes also increased with the reduced concentrations of RU486, which were the lowest when RU486 concentration was 10-7 mol/L, as compared with control and PQ exposure groups （P〈0.01 or P〈0.05）. The content of CAT enhanced with RU486 concentration, which was the highest when RU486 concentration was 10-7 mol/L, as compared with control and PQ exposure groups （P〈0.05）. But the change of MDA content was the contrary. Conclusion Nrf2 expression induced by RU486 can promote the balance of oxidation-antioxidation system in A549 cells and inhibit the inflammation and apoptosis factors, which has a protective effect on A549 cell injury induced by PQ.