沙库巴曲缬沙坦钠抑制血管紧张素Ⅱ诱导THP-1巨噬细胞基质金属蛋白酶-9的表达

Sacubitril valsartan sodium inhibits AngⅡ-induced MMP-9 expression in THP-1 macrophages via NF-κB pathway

目的探讨沙库巴曲缬沙坦钠(LCZ696)抑制血管紧张素Ⅱ(AngⅡ)诱导(THP)-1巨噬细胞基质金属蛋白酶(MMP)-9表达的机制。方法0.1μmol/L佛波酯(PMA)加至THP-1单核细胞48 h,将其诱导分化为THP-1巨噬细胞,随机分为4组:①PMA组,即对照组;②PMA+AngⅡ组(100μmol/L,1 h);③PMA+AngⅡ+LCZ696组(10μmol/L,1 h);④PMA+AngⅡ+二硫氨基甲酸肽吡咯烷(PDTC)组(100μmol/L,1 h)。用反转录-聚合酶链反应(RT-PCR)法检测MMP-9 mRNA的变化,用蛋白质印迹法检测胞质MMP-9蛋白的表达。结果与对照组相比,AngⅡ诱导的THP-1巨噬细胞MMP-9 mRNA表达上调(P<0.05),MMP-9蛋白量也增加(P<0.05),而使用LCZ696及PDTC后显著抑制MMP-9 mRNA的转录(P<0.05),MMP-9蛋白量也减少(P<0.05)。结论沙库巴曲缬沙坦钠可能通过核因子(NF)-κB信号途径抑制THP-1巨噬细胞MMP-9的表达。

Objective To investigate the mechanism underlying the inhibitory action of sacubitril valsartan sodium(LCZ696)against AngⅡ-induced MMP-9 expression in THP-1 macrophages.Methods 0.1μmol/L phorbol ester(PMA)was added to THP-1 monocytes for 48 h.The THP-1 monocytes were induced to differentiate into THP-1 macrophages,which were randomly divided into four groups:①PMA group,namely the control group;②PMA+AngⅡgroup(100μmol/L,1 h);③PMA+AngⅡ+LCZ696 group(10μmol/L,1 h);④PMA+AngⅡ+PDTC group(100μmol/L,1 h).RT-PCR was used to determine the change of MMP-9 mRNA expression,and Western blotting was used to determine the protein expression of cytoplasmic MMP-9.Results Compared with the control group,the mRNA expression level of AngⅡ-induced MMP-9 in THP-1 macrophages was up-regulated(P<0.05),and the protein expression level of MMP-9 also increased(P<0.05).The use of LCZ696 and PDTC significantly inhibited the mRNA transcription of MMP-9(P<0.05),and the protein expression level of MMP-9 decreased(P<0.05).Conclusion LCZ696 may inhibit the expression of MMP-9 in THP-1 macrophages via NF-κB pathway.