Tuo Qin-Hui, Liang Lei, Zhu Bing-Yang, Cao Xuan, Liao Duan-Fang
School of Life Science and Technology, University of South China, Western Changsheng Road #28, Hengyang, Hunan Province, China.
World J Gastroenterol. 2008 Jan 21;14(3):435-40. doi: 10.3748/wjg.14.435.
To study the effect of Daxx on cholesterol accumulation in hepatic cells.
Sprague Dawley (SD) rats were fed a normal or high fat diet for 6 wk, and serum lipids and Daxx expression of hepatic tissues were measured by immunoblot assays. HepG(2) cells were transfected with the pEGFP-C1/Daxx or pEGFP-C1 plasmid. Cells stably transfected with Daxx were identified by RT-PCR analysis. Total cholesterol levels were determined by high performance liquid chromatography. Activated-SREBP and caveolin-1 were assayed by western blotting.
Hepatic Daxx protein was higher in normal rats than in high fat diet-fed rats. Noticeable negative correlations were seen between Daxx and LDL-C (gamma = -7.56, P = 0.018), and between Daxx and TC (gamma = -9.07, P = 0.01), respectively. The total cholesterol of HepG(2)/GFP-Daxx cells was lower than that of control cells or HepG(2)/GFP cells (9.28 +/- 0.19 vs 14.36 +/- 4.45 or 13.94 +/- 2.62, both P < 0.05). Furthermore, in HepG(2)/GFP cells, the expression of activated SREBP was lower than that of control cells, whereas caveolin-1 expression was higher.
Overexpression of Daxx in HepG(2) cells decreased intracellular cholesterol accumulation, which might be associated with inhibition of SREBP activity and an increase in caveolin-1 expression.
研究死亡结构域相关蛋白(Daxx)对肝细胞内胆固醇蓄积的影响。
将Sprague Dawley(SD)大鼠分为正常饮食组和高脂饮食组,喂养6周,采用免疫印迹法检测血清脂质及肝组织中Daxx的表达。将pEGFP-C1/Daxx或pEGFP-C1质粒转染至HepG(2)细胞。通过逆转录-聚合酶链反应(RT-PCR)分析鉴定稳定转染Daxx的细胞。采用高效液相色谱法测定总胆固醇水平。通过蛋白质免疫印迹法检测活化的固醇调节元件结合蛋白(SREBP)和小窝蛋白-1(caveolin-1)。
正常大鼠肝组织中Daxx蛋白水平高于高脂饮食喂养的大鼠。Daxx与低密度脂蛋白胆固醇(LDL-C)(γ=-7.56,P=0.018)以及Daxx与总胆固醇(TC)(γ=-9.07,P=0.01)之间分别存在显著的负相关。HepG(2)/GFP-Daxx细胞的总胆固醇水平低于对照细胞或HepG(2)/GFP细胞(9.28±0.19 vs 14.36±4.45或13.94±2.62,均P<0.05)。此外,在HepG(2)/GFP细胞中,活化SREBP的表达低于对照细胞,而小窝蛋白-1的表达较高。
HepG(2)细胞中Daxx的过表达降低了细胞内胆固醇蓄积,这可能与抑制SREBP活性及小窝蛋白-1表达增加有关。