双氢睾酮对HaCaT细胞SREBP-1c表达的影响  

Effects of dihydrotestosterone on the expression of SREBP-lc in human HaCaT keratinocytes

作　　者：黄秋红[1] 周炳荣[1] 王丹[2] 郭娴菲[1] 骆丹[1] 

机构地区：[1]南京医科大学第一附属医院皮肤科,210029 [2]江苏省吴江市第一人民医院皮肤科

出　　处：《中华皮肤科杂志》2012年第10期735-738,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（30873407,81000700）

摘　　要：目的探讨双氢睾酮（DHT）在HaCaT细胞固醇调控元件结合蛋白-1c（SREBP-1c）表达中的作用。方法体外培养HaCaT细胞，分为4组，对照组不加任何刺激因素，DHT组分别加入3种不同浓度的DHT，LY294002十DHT组在加入50txmol／LP13K抑制剂（LY294002）预处理40min后加入100nmol／LDHT，PD98059＋DHT组即在加入50Ixmol／LMEK抑制剂（PD98059）预处理40rain后加入100nmol／LDHT。用实时定量PCR和Western印迹法检测DHT对HaCaT细胞SREBP-1cmRNA和蛋白表达的影响。Western印迹法检测DHT作用于HaCaT细胞后蛋白激酶B（AKT）、胞外信号调控激酶（ERK）、p38丝裂原活化蛋白激酶（p38MAPK）、c—Jun氨基末端激酶（JNK）的磷酸化情况。结果DHT可呈浓度依赖性上调HaCaT细胞SREBP-1cmRNA和蛋白的表达，并诱导AKT、ERK的磷酸化，但对p38、JNK的磷酸化无明显激活作用。LY294002预处理后HaCaT细胞SREBP-1cmRNA的表达较单纯DHT组明显降低（t=9．406，P〈0．05）；SREBP-1蛋白水平降为0．7113±0．0313，与单纯DHT组2．2577±0．0601比较，差异有统计学意义（t=39．498，P〈0．05）。而PD98059预处理后，SREBP-1emRNA和蛋白的表达与单纯DHT组比较，差异均无统计学意义（P〉0．05）。结论DHT可诱导HaCaT细胞SREBP-1cmRNA和蛋白的表达。Objective To evaluate the effects of dihydrotestosterone （DHT） on the expression of sterol regulatory element-binding protein-lc （SREBP-lc） in human HaCaT keratinocytes. Methods HaCaT cells were cultured in vitro and classified into 4 groups, i.e., control group receiving no treatment, DHT group treated with 3 different concentrations （10, 100, 1000 nmol/L） of DHT, LY294002 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the PI3K inhibitor LY294002 of 50 p.mol/L,PD98059 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the MEK inhibitor PD98059 of 50 ~mol/L. After another 24-hour culture, real time PCR and Western blot were carried out to detect the expression of SREBP-lc mRNA and protein in HaCaT cells, respectively. Western blot was also performed to determine the phosphorylation levels of protein kinase B （AKT）, extracellular signal-regulated kinase （ERK）, p38 mitogen- activated protein kinase and c-Jun N-terminal kinase （JNK） in the HaCaT cells. Results DHT could enhance the expression of SREBP-lc mRNA and protein in HaCaT cells in a concentration-dependent manner, and induce the phosphorylation of AKT and ERK, but not that of P38 or JNK. The expressions of SREBP-lc mRNA and protein were significantly decreased in HaCaT cells treated with LY294002 plus DHT （7.4780 ＋ 1.2638 vs. 21.6170 _＋ 2.2759, t --- 9.406, P 〈 0.05; 0.7113 ＋ 0.0313 vs. 2.2577 ＋ 0.0601, t = 39.498, P 〈 0.05）, but experienced no statistical changes in those treated with PD98059 and DHT（both P 〉 0.05）, compared with those treated with DHT only. Conclusion DHT can induce the expression of SREBP-lc mRNA and protein in HaCaT cells, likely via the PI3K/AKT signaling pathway.

关 键 词：双氢睾酮 HACAT细胞 胆固醇调节元件结合蛋白质1 信号转导 

分 类 号：R363[医药卫生—病理学]