【摘要】 目的探讨核因子-κB(NFκ-B)对外周血单核细胞基质金属蛋白酶9(MMP-9)mRNA表达的影响,以及二者与冠状动脉粥样硬化斑块不稳定性的关系。方法分离收集冠心病患者的外周血单核细胞,分别在不含血清的RPM I-1640中单独(空白对照组)或者加入氧化低密度脂蛋白(ox-LDL)(ox-LDL组)孵育24 h或先加PDTC(NF-κB抑制剂)(PDTC组)(10-5mol/L)培育1 h后加ox-LDL(40μg/m l)继续孵育24 h,收集单核细胞提取总RNA,并用RT-PCR方法测定MMP-9、组织金属蛋白酶抑制物1(TIMP-1)mRNA的表达,采用ELISA法测定上清液中MMP-9、TIMP-1浓度。结果细胞上清液中MMP-9蛋白及单核细胞MMP-9 mRNA表达增加(P<0.01)。与空白对照组比较,ox-LDL组PDTC表达减少(P<0.05);TIMP-1 mRNA表达及细胞上清液中TIMP-1蛋白含量在各组间无统计学差异。结论ox-LDL可诱导人单核细胞MMP-9表达增加,并且NFκ-B也参与了ox-LDL对MMP-9表达的促进作用,但对TIMP-1表达无影响。更多还原

【Abstract】 Objective To investigate the the influence of nuclear factor-kappaB on the expression of MMP-9 in monocytes and the relationship between the unstability of coronary atherosclerotic score and MMP-9 and nuclear factor-kappaB.Methods Circulating monocytes of patients with coronary heart disease were separated,identified,collected,divided into control group which were incubated in RPMI-1640 culture fluid,ox-LDL group which were incubated in RPMI-1640 with ox-LDL(40 μg/ml)and PDTC group which were pretreated with PDTC(10-5 mol/L).Then these cells were collected to isolate the total RNAs by the acid guanidium-thiocyanate-phenol-chloroform method and examined the expression of MMP-9 and TIMP-1 mRNA by RT-PCR.The supernatant liquid of cultured cells was also collected and used to determine the levels of MMP-9 and TIMP-1 by ELISA.Results Incubation of monocytes with ox-LDL for 24 hours markedly increased the expression MMP-9 mRNA(RT-PCR) and protein(ELISA)(P<0.01).Pretreatment of monocytes with PDTC prevented the expression MMP-9 mRNA in response to ox-LDL(P<0.05) and decreased the levels of MMP-9 protein(P<0.05).There was no significant deviation for the expression of TIMP-1 and protein in these groups.Conclusion ox-LDL can induce the expression of MMP-9 in monocytes.Nuclear factor-kappaB mediates the increasing expression of MMP-9 which is induced by ox-LDL in the procedure.While there is no effect on the expression of TIMP-1.更多还原