NF-κB对外周血单核细胞MMP-9 mRNA表达的影响  

Influences of nuclear factor-kappaB on the expression of MMP-9 in monocytes

作　　者：李如意[1] 李秀芳[1] 李英肖[1] 

机构地区：[1]河北省人民医院心脏中心,石家庄050051

出　　处：《山东医药》2010年第1期20-21,共2页Shandong Medical Journal

基　　金：河北省卫生厅医学科学研究重点课题(08016)

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。Objective To investigate the the influence of nuclear faetor-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 monocytcs 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 ceils 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 monocytcs 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.

关 键 词：核因子-ΚB 明胶酶B 组织金属蛋白酶抑制物-1 脂蛋白类 LDL 

分 类 号：R541.4[医药卫生—心血管疾病]