摘要
目的探讨非对称性二甲基精氨酸(ADMA)上调大鼠腹腔巨噬细胞的氧化低密度脂蛋白受体-1(LOX-1)表达是否受核因子-κB(NF-κB)的调控。方法分离培养大鼠腹腔巨噬细胞。实验分组:正常对照组,以含10%胎牛血清(FBS)DMEM培养液与巨噬细胞共孵育;氧化型低密度脂蛋白(oxLDL)组,在培养液中加oxLDL(50 mg/L);A+O组,在培养液中加ADMA(15μmol/L)和oxLDL(50 mg/L);P+A+O组,在培养液中加吡咯烷二硫代氨基甲酸盐(PDTC,25μmol/L)、ADMA(15μmol/L)o、xLDL(50 mg/L)。以实时荧光定量PCR(Real-time PCR)和Western blot分别检测LOX-1 mRNA和蛋白表达,以电泳迁移率变动分析(EMSA)和增强化学发光法(ECL)检测NF-κB活性。结果 oxLDL组与A+O组LOX-1 mRNA和蛋白表达比对照组明显增强,以A+O组尤为明显(均P<0.05);P+A+O组LOX-1mRNA和蛋白表达较A+O组降低(P<0.05)。与对照组相比,oxLDL组与A+O组NF-κB活性明显增强,以A+O组尤为明显(均P<0.05);P+A+O组NF-κB活性较A+O组降低(P<0.05)。相关分析:NF-κB活性与LOX-1 mRNA和蛋白表达量呈正相关(均为r=0.82,P<0.05)。结论 ADMA可能通过NF-κB途径增强LOX-1表达而促进巨噬细胞转化为泡沫细胞,促进动脉粥样硬化的发生发展。
Objective To investigate whether the expression of lectin-like oxidized low density lipoprotein receptor-1(LOX-1)in peritoneal macrophages of rats upregulated by asymmetric dimethylarginine(ADMA) is modulated by nuclear factor kappa B(NF-κB)pathway.Methods Wistar rat peritoneal macrophages were cultured and isolated,and divided into four groups:①Control group:macrophages incubated in DMEM media with 10% fetal bovine serum(FBS)without any intervention;②Oxidized low density lipoprotein(oxLDL)group:macrophages incubated in DMEM media with 10% FBS and oxLDL(50 mg/L);③A+O group:macrophages incubated in DMEM media with 10% FBS,ADMA(15 μmol/L)and oxLDL(50 mg/L);④P+A+O group:macrophages incubated in DMEM media with 10% FBS,PDTC(25 μmol/L),ADMA(15 μmol/L)and oxLDL(50 mg/L).The mRNA and protein expression of LOX-1 was detected by real-time fluorescent quantitative PCR(real-time PCR)and Western blot respectively.The activity of NF-κB was measured by electrophorestic mobility shift assay(EMSA)and enhanced chemiluminescent(ECL)technique.Results ①As compared with control group,the mRNA and protein expression of LOX-1 was significantly increased in oxLDL group and A+O group,especially in A+O group(P0.05),and the mRNA and protein expression of LOX-1 in P+A+O group was significantly decreased as compared with that in A+O group(P0.05);②The NF-κB activity in both A+O group and oxLDL group was obvionsly higher than that in control group,especially in A+O group(P0.05),and the NF-κB activity in P+A+O group was significantly lower than that in A+O group(P0.05);③Correlation analysis revealed that the activity of NF-κB was positively correlated with the mRNA and protein expression of LOX-1(r=0.82,P0.05).Conclusion ADMA upregulated the LOX-1 expression through the NF-κB activity pathway in rat peritoneal macrophages,which might be one of mechanisms of foam cell formation and atherosclerosis.
出处
《华中科技大学学报(医学版)》
CAS
CSCD
北大核心
2010年第6期771-774,779,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong