Inhibition of superoxide dismutase induces collagen production in cardiac fibroblasts
Inhibition of superoxide dismutase induces collagen production in cardiac fibroblasts
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DOI：
10.1038/ajh.2008.242
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发表时间：
2008-10-01
影响因子：
3.2
通讯作者：
Fagard, Robert
中科院分区：
文献类型：
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作者：
Lijnen, Paul;Petrov, Victor;Fagard, Robert
本研究的目的是确定用二乙基二硫代氨基甲酸（DETC）抑制超氧化物歧化酶（SOD）是否会影响对照和血管紧张素II（ANG II）处理的心脏成纤维细胞中胶原的产生、I型和III型胶原以及纤维连接蛋白的mRNA和蛋白表达。方法将心肌成纤维细胞培养至汇合，在无血清Dulbecco改良Eagle培养基中孵育24 h，与（外）测试抑制剂预孵育1小时，并与（外）ANG II进一步孵育（1 μ mol/L）作用24 h，丹参总黄酮呈剂量依赖性地抑制心肌成纤维细胞CuZn-SOD活性。超氧阴离子的产生增加DETC和减少tempol在控制和ANG II处理的成纤维细胞。DETC也减少了细胞内产生的活性氧（ROS）（如H2 O2，羟基自由基，氢过氧化物）在控制和ANG II处理的成纤维细胞，而tempol减少ROS生产仅在ANG II处理的成纤维细胞。ANG Ⅱ和DETC刺激成纤维细胞胶原合成及Ⅰ型胶原和纤维连接蛋白含量。SOD模拟物tempol和EUK-8以及PEG-SOD减少胶原的产生。ANG II和DETC刺激金属蛋白酶组织抑制剂-I（TIMP-1）和TIMP-2的水平，而tempol降低TIMP-2的含量在控制和ANG II处理的成纤维细胞。基质金属蛋白酶（MMP）-1水平在ANG Ⅱ和DETC作用下降低，在Tempol作用下升高。结论SOD和ROS在心肌成纤维细胞胶原积累中起重要作用。
BACKGROUNDThe aim of this study was to determine whether inhibition of superoxide dismutase (SOD) with diethyldithiocarbamic acid (DETC) could affect the collagen production, the mRNA and protein expression of collagen types I and III, and fibronectin in control and angiotensin II (ANG II)-treated cardiac fibroblasts. Its effect was compared with the SOD mimetics tempol and EUK-8 and with polyethyleneglycol (PEG)-SOD.METHODSCardiac fibroblasts were cultured to confluence, incubated in serum-free Dulbecco's modified Eagle's medium for 24 h, preincubated with(out) the tested inhibitors for 1 h and further incubated with(out) ANG II (1 mu mol/l) for 24 h.RESULTSDETC dose-dependently inhibited the activity of CuZn-SOD in cardiac fibroblasts. Superoxide anion production was increased by DETC and decreased by tempol in control and ANG II-treated fibroblasts. DETC also reduced the intracellular generation of reactive oxygen species (ROS) (such as H2O2, hydroxyl radicals, hydroperoxides) in control and ANG II-treated fibroblasts, whereas tempol reduced the ROS production only in ANG II-treated fibroblasts. ANG II and DETC stimulated the collagen production and the collagen I and fibronectin content in fibroblasts. The SOD mimetics tempol and EUK-8 as well as PEG-SOD reduced the Collagen production. ANG II and DETC stimulated the tissue inhibitor of metalloproteinase-I (TIMP-1) and TIMP-2 levels, whereas tempol decreased the TIMP-2 content in control and ANG II-treated fibroblasts. Matrix metalloproteinase (MMP)-1 level was reduced by ANG II and DETC and increased by tempol.CONCLUSIONThese data suggest a vital role of SOD and the formed ROS in the accumulation of Collagen in cardiac fibroblasts.