一氧化氮在白细胞介素-1β诱导的基质金属蛋白酶-2活性上调中的作用  

Effect of nitric oxide on up-regulation of matrix metalloproteinase-2 activity induced by IL-1β

作　　者：黄进宇[1] 程刚[2] 翁少翔[2] 单江[2] 

机构地区：[1]杭州市红十字会医院心内科,浙江省杭州310003 [2]浙江大学医学院附属第二医院

出　　处：《中华急诊医学杂志》2005年第5期394-397,共4页Chinese Journal of Emergency Medicine

摘　　要：目的研究一氧化氮(NO)在白细胞介素-1β(IL-1β)诱导的血管平滑肌细胞(VSMC)分泌的基质金属蛋白酶2(MMP2)活性上调中的作用,探讨IL1β调节MMP2活性的可能机制。方法组织贴块法培养VSMC。Zymography检测MMP2活性,Western印迹法分析MMP2蛋白表达水平,硝酸还原酶法及黄嘌呤氧化酶法分别测定细胞培养上清中NO及丙二醛(MDA)水平。结果IL-1β刺激后,细胞培养液的MMP2活性上升为对照组的(5.67±1.09)倍(P<0.05),而IL-1β氨基胍(AG)组MMP2活性则为对照组的(7.76±1.24)倍(P<0.05);IL-1β刺激后,细胞培养上清中MMP2蛋白表达量上升为对照组的(3.26±0.32)倍(P<0.05),而IL-1β+AG组MMP2蛋白表达量则为对照组的(4.35±0.48)倍(P<0.05);对照组细胞培养上清中NO、MDA水平分别为(2.89±0.81)μmol/g.protein、(8.85±2.80)μmol/g.protein,II-1β刺激后细胞培养上清中N0、MDA水平上升为(116.85±20.29)μmol/g.protein、(24.91±6.24)μmol/g.protein,IL1β+AG组NO水平[(36.78±7.18)μmol/g.protein]较Il-1β组降低(P<0.05),而MDA水平[(55.75±17.49)μmol/g.protein]则较IL1β组上升(P<0.05)。结论IL1β刺激VSMC后释放的NO可能通过抑制IL1β诱导的超氧化物的大量产生而对MMP2活性起抑制作用。Objective To investigate the effect of nitric oxide on up-regulation of matrix metalloproteinase-2 activity induced by IL-1β.Methods The activity of MMP-2 was measured by gelatin zymography,the protein levels of MMP-2 were examined by using Western blot technique,and the concentrations of NO and MDA were measured according to the kit reference.Results Stimulation of vascular smooth muscle cell(VSMC) with IL-1β significantly increased MMP-2 activity and protein expression of media(P<0.05),pretreatment with IL-1β and aminoguanidine,MMP-2 activity and protein expression of media were lower(P<0.05).Stimulated with IL-1β significantly increased NO and MDA content of media(P<0.05);pretreatment with IL-1β and Aminoguanidine,NO content was lower(P<0.05),but MDA level was higher(P<0.05).Conclusion After stimulated by IL-1β,NO has the inhibitory effect on MMP-2.

关 键 词：白细胞介素-1Β 一氧化氮 基质金属蛋白酶-2(MMP-2) 活性 protein 上调 Western印迹法 MMP-2蛋白 IL-1β 丙二醛(MDA) 血管平滑肌细胞 白细胞介素1β 黄嘌呤氧化酶法 培养上清 蛋白表达水平 硝酸还原酶法 VSMC 组织贴块法 

分 类 号：R543.3[医药卫生—心血管疾病]