麻黄碱对TNF-α诱导的HaCaT细胞增殖、凋亡及细胞周期的影响

The effect of ephedrine on the proliferation,apoptosis and cell cycle in HaCaT cells induced by tumor necrosis factor alpha

目的研究麻黄碱对TNF-α诱导的永生化角质形成细胞HaCaT细胞的增殖、凋亡及细胞周期的影响。方法倒置显微镜下观察细胞生长情况,MTT法检测TNF-α诱导下HaCaT细胞增殖情况、800μg/ml麻黄碱对TNF-α诱导的HaCaT细胞增殖的影响。在流式细胞仪上使用Annexin-V/PI和PI单标试剂盒分别检测HaCaT细胞的凋亡及细胞周期。结果 25ng/ml的TNF-α能明显促进HaCaT细胞的增殖。800μg/ml麻黄碱作用于TNF-α诱导增殖下的HaCaT细胞24 h后,悬浮细胞数目变多、贴壁细胞分布较前稀疏,细胞内空洞变多。MTT显示800μg/ml麻黄碱能够明显抑制TNF-α诱导的细胞增殖,与空白对照组比价有显著性差异(P<0.05)。流式细胞仪检测结果显示:麻黄碱使TNF-α诱导下的HaCaT细胞G1期明显延长,但对细胞凋亡未产生影响。结论一定浓度的麻黄碱可以抑制TNF-α对HaCaT细胞的促增殖作用,并使诱导后的细胞停滞在G1期。

Objective To evaluate the effect of ephedrine on the proliferation,apoptosis and cell cycle in immortalized keratinocytes（ HaCaT cells） induced by tumor necrosis factor alpha（ TNF-α）. Methods HaCaT cells were observed through inverse microscope. The proliferation of cells induced by TNF-a or combination of TNF-α and 800ug/ml ephedrine were measured with methyl thiazolyl tetrazolium（ MTT） assay. Flow cytometry using Annexin-V/propidium iodine（ PI） or PI were respectively performed to assess the apoptosis and the cell cycle of HaCaT cells. Results The proliferation activity of HaCaT cells were obviously promoted by 25ng/ml TNF-α. After 24 hours stimulated by combination of 800μg/ml ephedrine with the TNF-α,more suspension cells were found,the adherent cells were sparse and more empty holes were observed. MTT assay showed that ephedrine of 800μg/ml effectively inhibited the proliferation of HaCaT cells induced by TNF-α of 25 ng/ml and had significant difference compared with the blank control group（ P〈0. 05）. Flow cytometry instrument testing showed that ephedrine could increase the cell cycle of G1 phase ratio,but it had no effect on cell apoptosis. Conclusion A certain concentration of ephedrine could suppress the promotive effect of TNF-α on the proliferation of HaCaT cells and block the cells in G1 phase.