Mohammadi Abbas, Najar Ahmad Gholamhoseinian, Khoshi Amirhosein
Department of Biochemistry, Physiology Research Center, Kerman, Iran.
Department of Biochemistry, Kerman University of Medical Sciences, Kerman, Iran.
Adv Biomed Res. 2014 Jan 9;3:22. doi: 10.4103/2277-9175.124661. eCollection 2014.
Increased apolipoprotein B100 (apo B) and decreased apolipoprotein A-I (apo A-I) production are important risk factors in atherosclerosis. Urotensin II (UII), as the most potent vasoconstrictor in human, is related with hypertension and probably atherosclerosis. Because of the relationship between the hypertension and lipoprotein metabolism in atherosclerosis, the aim of this study was to test the effect of urotensin II on apo B and apo A-I expression in hepatic (HepG2) cell line.
HepG2 cells were treated with 10, 50, 100, and 200 nmol/L of urotensin II (n = 6). Relative apo B and apo A-I messenger RNA (mRNA) levels in conditioned media, normalized to glyceraldehyde-3-phosphate dehydrogenase, were measured with quantitative real-time polymerase chain reaction method. In addition, apo B and apo A-I levels were also estimated and compared with the controls using the western blotting method. Data were analyzed statistically by ANOVA and non-parametric tests.
The apo B mRNA levels were not increased significantly following the treatment with UII. However, apo B protein levels were increased significantly after the treatment with urotensin II, especially at 100 and 200 nmol/L. The apo A-I mRNA and protein levels in conditioned media also were not significantly changed. However, there was a significant decrease in apo A-I mRNA and protein levels at 200 nM UII.
UII might increase apo B at protein level probably through participating factors in its synthesis and/ or stability/degradation. In addition, UII may have decreasing effect at more than 200 nM concentrations on apo A-I.
载脂蛋白B100(apo B)升高和载脂蛋白A-I(apo A-I)生成减少是动脉粥样硬化的重要危险因素。尾加压素II(UII)作为人体内最有效的血管收缩剂,与高血压以及可能的动脉粥样硬化相关。鉴于高血压与动脉粥样硬化中脂蛋白代谢的关系,本研究旨在检测尾加压素II对肝(HepG2)细胞系中apo B和apo A-I表达的影响。
用10、50、100和200 nmol/L的尾加压素II处理HepG2细胞(n = 6)。用定量实时聚合酶链反应法测量条件培养基中相对apo B和apo A-I信使核糖核酸(mRNA)水平,并以甘油醛-3-磷酸脱氢酶进行标准化。此外,还用蛋白质印迹法估计apo B和apo A-I水平并与对照组进行比较。数据采用方差分析和非参数检验进行统计学分析。
用UII处理后,apo B mRNA水平未显著升高。然而,用尾加压素II处理后,apo B蛋白水平显著升高,尤其是在100和200 nmol/L时。条件培养基中的apo A-I mRNA和蛋白水平也没有显著变化。然而,在200 nM UII时,apo A-I mRNA和蛋白水平有显著下降。
UII可能通过参与其合成和/或稳定性/降解的因素在蛋白质水平上增加apo B。此外,UII在浓度超过200 nM时可能对apo A-I有降低作用。