全反式维A酸对HaCaT细胞Arnt及Ahr表达的影响

The Effect of All-trans Retinoic Acid on Expression of Arnt and Ahr in HaCaT Cell

目的研究全反式维A酸作用下,皮肤屏障相关因子Arnt及Ahr的变化,以明确维A酸对皮肤屏障相关因子的影响及可能分子机制。方法 1μmol/L的全反式维A酸作用人角质形成细胞HaCaT 36h后,分别通过基因芯片、实时定量PCR法、免疫印迹法来检测Arnt及Ahr在基因及蛋白水平的变化。结果人角质形成细胞HaCaT在全反式维A酸作用36h后,基因表达谱差异显示:Arnt基因下调3.95倍,Ahr基因上调2.13倍;实时定量PCR法结果显示,与对照组比较,加药组Arnt mRNA表达量下降(0.35±0.03)倍(P<0.05),Ahr mRNA表达量升高(2.47±0.07)倍(P<0.05);免疫印迹结果显示:加药组Arnt蛋白明显降低而Ahr蛋白明显升高,差异均具有统计学意义。结论全反式维A酸作用于HaCaT细胞在基因及蛋白水平均可引起Arnt的表达降低和Ahr的表达升高,提示Arnt及Ahr在全反式维A酸对表皮细胞影响过程中发挥不同的生物学作用。

Objective To investigate whether skin barrier related molecules Arnt and Ahr was affected by ATRA in HaCaT cell. Methods Pre-treated the HaCaT cells with ATRA （ 1 p^mol/L） for 36h were used to detect the expression of Arnt and Ahr by gene microarray technology, Western blot and Reahime-PCR assay. Results Comparing with the control group, the level of Arnt gene was suppressed by 3.95 times while Ahr gene was elevated 2. 13 times using gene microarray detection after treatment by ATRA for 36h; the level of Arnt mRNA was down-regulated （0. 35 ± 0. 03 ） and the Ahr mRNA was up-regulated （2. 47 ± 0. 07 ） by qRTPCR（P 〈 0. 05）. Western blot results were consistent with above detection. Conclusion The expression of Amt is down-regulated in gene and protein level while Ahr shows opposite results in cultured HaCaT cells treated by ATRA （lμmol/L） for 36h. These two molecules may play different roles in the skin biology affected by ATRA.