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目的探讨非吞噬细胞氧化酶4(NOX4)在转化生长因子β(TGF-β)诱导A549细胞迁移中的作用。方法实验分组:TGF-β(刺激)组,空白对照组,二甲苯基碘(DPI,NOX4抑制剂)组,TGF-β+DPI组,二甲基亚砜(DMSO)(溶剂对照)组。流式细胞仪检测ROS表达;Western blot检测NOX4、snail、E-cadherin蛋白表达;划痕实验检测A549细胞迁移能力。结果经Quantity One软件定量后,TGF-β组NOX4表达为:1.80±0.07,TGF-β+DPI组为0.49±0.03(F=327.071,P<0.001);上皮间质转化相关蛋白:TGF-β组snail蛋白为9.0±0.6,TGF-β+DPI组为1.8±0.3(F=119.097,P<0.001);TGF-β组E-cadherin蛋白为0.5±0.1,TGF-β+DPI组为3.3±0.3(F=71.063,P<0.001);以上结果表明DPI可以抑制A549细胞NOX4表达且可以抑制其EMT进程。TGF-β+DPI组与TGF-β组比较,划痕愈合率降低(F=33.899,P<0.001);表明DPI可以抑制A549细胞的迁移。结论 DPI抑制NOX4的表达后,TGF-β诱导的A549细胞迁移被抑制,并且与TGF-β诱导的上皮间质转化进程有关。
Objective To investigate the role of non-phagocytic oxidase 4 (NOX4) in the migration of A549 cells induced by transforming growth factor β (TGF-β). Methods The experimental groups were divided into three groups: TGF-β (stimulus) group, blank control group, xylenyl iodide (DPI, NOX4 inhibitor) group, TGF-β + DPI group and DMSO (solvent control group) The expression of ROS was detected by flow cytometry. The protein expression of NOX4, snail and E-cadherin were detected by Western blot. Scratch assay was used to detect the migration ability of A549 cells. Results Quantitative Quantity One software showed that the expression of NOX4 in TGF-β group was 1.80 ± 0.07 and that in TGF-β + DPI group was 0.49 ± 0.03 (F = 327.071, P <0.001) The protein content of snail was 9.0 ± 0.6 in the TGF-β + DPI group and 1.8 ± 0.3 in the TGF-β + DPI group (F = 119.097, P <0.001) 0.3 (F = 71.063, P <0.001). These results indicated that DPI can inhibit the expression of NOX4 in A549 cells and inhibit its EMT progression. Scratch healing rate was lower in TGF-β + DPI group than in TGF-β group (F = 33.899, P <0.001); DPI could inhibit A549 cell migration. Conclusion DPI inhibits the expression of NOX4, TGF-β-induced A549 cell migration is inhibited, and TGF-β-induced epithelial mesenchymal transition process.