雌激素对体外培养人毛乳头细胞生物钟基因表达的影响

Effect of estrogen on expression of circadian clock genes in human dermal papilla cells in vitro

目的:探讨雌激素雌二醇(E2)对于体外培养的原代毛乳头细胞(DPC)的生物钟基因活性的影响。方法:体外分离培养6个月胎龄畸形引产胎儿头皮毛乳头细胞,2周后同步所有细胞生物钟活性,随机分为雌激素E2实验组和对照组。24 h中每间隔6 h提取一次总RNA,采用荧光定量PCR方法检测细胞中Clock、Bmal1、Per1、Per2、Per3等主要生物钟基因的表达量。结果:贴壁后的原代DPCs呈放射状向外生长,单个细胞形态呈长梭形。无论雌激素E2实验组和空白对照组,DPCs生物钟基因均呈节律性表达。雌激素E2处理后,相对于对照组,实验组生物钟基因(Clock, Bmal1, Per1, Per2)的表达量显著增加(P<0.05)。结论:雌激素E2显著调节体外培养人毛乳头细胞生物钟基因的表达水平,其生物学意义值得深入研究。

Objective:To explore the influence of estrogen(E2)on the expression of circadian clock genes in cultured primary human dermal papilla cells(DPCs).Methods:DPCs were obtained from the scalp tissue of a six-month-old nonviable fetus.Two weeks later,the DPCs were synchronized and randomly divided into the experimental group treated with E2 and the controls.RNA was extracted every 6 hours over a 24-hour period.Quantitative polymerase chain reaction(RT-PCR)was used to assess the transcriptional levels of core genes,including Clock,Bmal1,Per1,Per2 and Per3.Results:The primary DPCs displayed a spread-out appearance,while individual cell appeared in long fusiform shape.Both controls and E2-treated DPCs showed rhythmic expression of clock genes.The transcript levels of core clock genes,including Clock,Bmal1,Per1 and Per2,were up-regulated significantly in DPCs treated with E2 for 24 hours as compared to the vehicle controls(P<0.05).Conclusions:Estrogen could modulate the expression of circadian clock genes in DPCs.Its biological significance is worthy of further exploration.