丙烯醛对肺上皮细胞增殖活力及生物钟基因Per1蛋白表达的影响

Effects of Acrolein on the Proliferation of and Per1 Gene Expression in Pulmonary Epithelial Cells

目的研究丙烯醛对肺上皮细胞增殖活力的影响及其可能机制。方法选用A549和MLE15两种肺上皮细胞株作为模型,分别用PBS(对照组)和80μmol/L丙烯醛处理30 min。采用CCK-8试剂盒检测丙烯醛处理结束恢复培养12 h、24 h、36 h和48 h时细胞的增殖活力,Western blot检测丙烯醛处理结束恢复培养24 h时生物钟基因Per1的蛋白表达水平。检测在恢复培养阶段加入10 mg/L转化生长因子-β(transforming growth factor-β,TGF-β)处理对丙烯醛处理后的肺上皮细胞增殖活力及Per1蛋白表达的影响。结果80μmol/L丙烯醛刺激30 min可导致A549细胞及MLE15细胞增殖活力明显降低以及细胞Per1蛋白表达下降(P<0.05)。丙烯醛处理结束以后,在细胞恢复培养过程中添加10 mg/L TGF-β未明显改变丙烯醛诱导的细胞增殖活力的降低,但是肺上皮细胞中Per1蛋白的表达水平高于未添加TGF-β的丙烯醛处理组(P<0.05)。结论丙烯醛会导致肺上皮细胞增殖活力以及Per1表达量降低,TGF-β虽然不能逆转丙烯醛导致的细胞增殖活力的下降,但可以使Per1的表达得到一定程度的恢复。

Objective To investigate the effect of acrolein on the proliferation of pulmonary epithelial cells and its possible mechanism.Methods Two strains of pulmonary epithelial cells,A549 cells and MLE15 cells,were used as in vitro models of pulmonary epithelial cell,and were treated with 80μmol/L acrolein or phosphate buffer saline(PBS)as the control.The proliferation of pulmonary epithelial cells were determined with CCK-8 kit after cell culturing resumed for12 h,24 h,36 h and 48 h post acrolein treatment,and the expression of period circadian regulator gene 1(Per1)was examined using Western blot test 24 h after acrolein treatment.In addition,after acrolein treatment,the cells were restored with transforming growth factor-β(TGF-β)added in the medium,and the cell proliferation and the expression of Per1 protein were also examined.Results The proliferation of A549 cells and MLE15 cells decreased significantly after being treated with 80μmol/L acrolein for 30 min,and the expression of Per1 protein was also downregulated significantly(P<0.05).The addition of TGF-βafter acrolein treatment did not significantly change the reduction in cell proliferation caused by acrolein,but the expression of Per1 protein in pulmonary epithelial cells was significantly higher than that in cells restored without TGF-β(P<0.05).Conclusion Acrolein treatment resulted in the decreased proliferation of pulmonary epithelial cells and the Per1 expression in pulmonary epithelial cells.Although TGF-βaddition did not reverse the reduction of cell proliferation after acrolein treatment,the Per1 expression levels were recovered to a certain extent compared to that in cells restored in medium without TGF-βafter acrolein treatment.