髓过氧化物酶对THP-1巨噬细胞活性及MMP-9、TIMP-1表达的影响  

Effect of myeloperoxidase on activity of THP-1 macrophage and expression of MMP-9 and TIMP-1

作　　者：孙定军[1] 邢波[1] 陈漠水[1] 林德洪[1] 张福伟 SUN Dingjun;XING Bo;CHEN Moshui;LIN Dehong;ZHANG Fuwei(Department of Cardiology,Haikou People′s Hospital and Haikou Hospital Affiliated to Xiangya Medical College, Haikou 570208, China)

机构地区：[1]海口市人民医院暨湘雅医学院附属海口医院心内科,海口570208

出　　处：《新疆医科大学学报》2020年第4期425-429,共5页Journal of Xinjiang Medical University

基　　金：海南省卫生计生行业科研项目(18A200159)。

摘　　要：目的探讨髓过氧化物酶(MPO)对单核巨噬细胞(THP-1)、活性及基质金属蛋白酶-9(MMP-9)、组织金属蛋白酶抑制剂1(TIMP-1)表达的影响。方法2018年12月-2019年3月于海口市人民医院暨湘雅医学院附属海口医院中心实验室进行实验,设THP-1为对照组、MPO低、中、高剂量组(MPO终浓度分别为1.5、3.0、6.0μ/L),各组每孔设6个平行样,培养72 h。试验结束后,MTT法测定细胞活力,流式细胞仪测定细胞凋亡水平,实时荧光逆转录法(RT-PCR)及蛋白印迹法(WESTERN-BLOT)测定细胞MMP-9、TIMP-1基因和蛋白水平。结果与对照组比较,MPO低、中、高剂量组OD值、存活率水平明显升高(P<0.05),且随着MPO剂量逐渐增加,各组OD值、存活率逐渐升高(P<0.05)。与对照组比较,MPO低、中、高剂量组凋亡率水平降低(P<0.05),且随着MPO剂量逐渐增加,各组凋亡率逐渐降低(P<0.05)。与对照组比较,MPO低、中、高剂量组MMP-9 mRNA和蛋白表达水平明显升高(P<0.05),TIMP-1mRNA和蛋白表达水平明显降低(P<0.05);且随着MPO剂量逐渐增加,各组MMP-9 mRNA和蛋白表达水平逐渐升高(P<0.05),而TIMP-1mRNA和蛋白表达水平逐渐降低(P<0.05)。结论MPO可促进巨噬细胞增殖,抑制其凋亡,加重动脉粥样硬化病变进展;其机制与MPO可促进巨噬细胞MMP-9 mRNA和蛋白表达,抑制TIMP-1mRNA和蛋白表达有关。Objective To investigate the effect of myeloperoxidase on activity of THP-1 macrophage and expression of matrix metalloproteinase-9(MMP-9)and tissue metalloproteinase inhibitor-1(TIMP-1).MethodsSet up THP-1 macrophage control group,low(1.5μ/L),medium(3.0μ/L)and high(6.0μ/L)dose groups of myeloperoxidase(MPO).Six parallel samples were collected in every group for 72 hours.After the experiment,cell viability was measured by MTT,apoptotic level was measured by flow cytometry,and MMP-9,TIMP-1 gene and protein levels were determined by PCR and Western Blotting.Res-ultsCompared with the control group,the OD value and survival rate of the low,medium and high dose groups of MPO were significantly increased dose-dependently(P<0.05).Compared with the control group,the levels of apoptotic rate in the low,medium,and high dose groups of MPO decreased dose-dependently(P<0.05).Compared with the control group,the expression levels of MMP-9 mRNA and protein in the low,medium and high dose groups of MPO were significantly increased dose-dependently(P<0.05),while the expression levels of TIMP-1 mRNA and protein were significantly decreased dose-dependently(P<0.05).ConclusionMPO can promote macrophage proliferation,inhibit apoptosis and aggravate atherosclerosis.The mechanism of MPO is related to promote the expression of MMP-9 mRNA and protein in macrophages,and inhibit the expression of TIMP-1 mRNA and protein.

关 键 词：MPO THP-1巨噬细胞 基质金属蛋白酶-9 组织金属蛋白酶抑制剂-1 

分 类 号：R543[医药卫生—心血管疾病]