NF-κB对冠心病患者外周血单核细胞LOX-1 mRNA和蛋白表达的影响

目的探讨核因子-κB（NF-κB）对冠心病患者外周血单核细胞凝集素样氧化低密度脂蛋白受体-1（LOX-1）mRNA表达的影响。方法分离收集冠心病患者外周血单核细胞,分为3组:对照组在不含血清的RPMI-1640中孵育24h;ox-LDL组在不含血清的RPMI-1640中加ox-LDL（40μg/ml）孵育24h;PDTC（NF-κB抑制剂）组:先加PDTC(10^-5mol/L)培育1h之后加ox-LDL（40μg/ml）继续孵育24h,之后收集单核细胞及细胞上清液,采用异硫氰酸胍—酚—氯仿抽提法提取总RNA,扩增目的基因LOX-l,并采用ELISA法测定上清液中sLOX-1蛋白浓度。结果与对照组比较,ox-LDL组单核细胞LOX-1 mRNA表达增加（0.304±0.047vs0.813±0.131,P<0.05）,细胞上清液中sLOX-1蛋白含量（ng/ml）增加（7.277±1.979 vs 16.517±2.064,P<0.05）;PDTC组单核细胞LOX-1 mRNA表达及细胞上清液中sLOX-1蛋白含量均增高（分别为0.502±0.140和11.997±1.757,P<0.05）。与ox-LDL组相比,PDTC组单核细胞LOX-1 mRNA表达减少,细胞上清液中sLOX-1蛋白含量显著减少（P<0.05）。结论 ox-LDL可诱导人单核细胞LOX-1表达增加,NF-κB也参与了ox-LDL对LOX-1表达并有促进作用。

The influence of nuclear factor-kappaB on the expression of LOX-1 mRNA and protein in monocytes

Objective To investigate the influence of nuclear factor-kappaB（NF-κB） on the expression of LOX-1 in monocytes in patients with coronary heart disease（CHD）.Methods Circulating monocytes of patients with CHD were separated, identified,collected,divided into 3 groups,control group were incubated in RPMI-1640 culture fluid,ox-LDL group were incubated in RPMI-1640 with ox-LDL（40μg/ml）and PDTC group were pretreaf with PDTC(10^-5mol/L).Then these cells were collected to isolate the total RNA by the acid guanidium-thiocyanate-phenol-chloroform method and examined the expression of LOX-1 mRNA by RT-PCR.The supernatant liquid of cultured cells was also collected and used to determine the levels of LOX-1 protein by ELISA.Results Compared with control group,the monocyte expression of LOX-1 mRNA increased significantly in ox-LDL group（0.304±0.047 vs 0.813±0.131,P<0.05）,sLOX-1 protein（ng/ml） in supernatant liquid increased significantly（7.277±1.979 vs 16.517±2.064,P<0.05）,the LOX-1 mRNA and sLOX-1 protein increased in PDTC group too（0.502±0.140,11.997±1.757,P<0.05）.Compared with those in ox-LDL group,the expression of LOX- 1 mRNA and content of sLOX-1 protein in supernatant decreased significantly in PDTC group（P<0.05）.Conclusion ox-LDL can induce the expression of LOX-1 in monocytes,NF-κB mediates the increasing expression of LOX-1 which is induced by ox-LDL in the procedure.