摘要
目的观察非对称性二甲基精氨酸(asymmetric dimethylarginine,ADMA)对培养的大鼠腹腔巨噬细胞诱导型一氧化氮合酶(iNOS)表达的影响,探讨其在脂质沉积中的作用及机制。方法分组①正常对照组以PBS缓冲液与巨噬细胞共孵育,②氧化型低密度脂蛋白(oxLDL)组在巨噬细胞培养液中加oxLDL(50mg/L),③O+A组在巨噬细胞培养液中加oxLDL(50mg/L)和ADMA(20μmol/L),④O+A+L组在培养液中加ox-LDL(50mg/L)、ADMA(20μmol/L)、L-Arg(1.2mmol/L)。以硝酸还原酶法和化学比色法分别测定培养液中一氧化氮(NO)含量与iNOS活力,以RT-PCR和Western Blotting分别检测iNOSmRNA和蛋白表达。结果①OxLDL组与O+A组NO含量降低,iNOS活力升高,且以O+A组明显,与正常对照组比较差异均有统计学意义(均P<0.05);NO与iNOS呈负相关(r=-0.697,P<0.05)。②O+A组iNOS mRNA和蛋白表达增强,与正常对照组相比差异有统计学意义(P<0.05或P<0.01)。结论ADMA抑制大鼠腹腔巨噬细胞合成NO,增强iNOS基因与蛋白表达,可能是ADMA促使巨噬细胞转变为泡沫细胞、促进动脉粥样硬化发生发展的机制之一。
Objective：To observe the effects of asymmetric dimethylarginine （ADMA） on inducible nitric oxidize synthase（iNOS） expression of cultured celiac macrophages in rats for investigating the significance of ADMA in foam cell formation. Method：Groups： （1）Control group： macrophages and PBS coincubated, （2）oxLDL group： macrophages and oxidized low density lipoprotein （oxLDL, 50 mg/L） coincubated, （3）O＋ A group： macrophages, oxLDL（50 mg/L） and ADMA （20 μmol/L） incubated together, （4） O＋ A＋ L group ： macrophages , oxLDL （50 mg/L）, ADMA（20 μmol/L）and L-Arginine （L-Arg, 1.2 mmol/L） incubated together. NO and iNOS were measured by nitrate reductase and chem-colorimetry respectively. The expressions of iNOS mRNA and protein were detected by reverse transcription polymerase chain reaction （RT-PCR） and Western Blotting. Result：（1） NO level decreased and iNOS activity increased significantly in oxLDL group and O＋A group, compared with control group, especially in O＋A group （P〈0. 05） ; NO was correlated with iNOS negatively （r= -0. 697, P〈0. 05）. （2）The expression of iNOS mRNA and protein in O＋A group were extremely higher than those in control group （P〈 0.05 or P〈0. 01）. Conclusion： ADMA depresses NO production and enhances iNOS expression of cultured celiac macrophages in rats, which might be one of the mechanisms of foam cell formation and atherogenesis.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2007年第4期280-283,共4页
Journal of Clinical Cardiology
