The role of chemically induced glutathione and glutathione S-transferase in protecting against 4-hydroxy-2-nonenal-mediated cytotoxicity in vascular smooth muscle cells.
The role of chemically induced glutathione and glutathione S-transferase in protecting against 4-hydroxy-2-nonenal-mediated cytotoxicity in vascular smooth muscle cells.
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DOI：
10.1385/ct:3:2:165
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发表时间：
2003-01-01
影响因子：
3.2
通讯作者：
Li, Yunbo
中科院分区：
文献类型：
--
作者：
Cao, Zhuoxiao;Hardej, Diane;Li, Yunbo
4-羟基-2-壬烯醛(HNE)被认为与动脉粥样硬化的发病机制有关。HNE的主要代谢途径之一是在谷胱甘肽S转移酶(GST)的催化下与谷胱甘肽(GSH)结合。在本研究中，我们研究了~3H-1，2-二硫醇-3-硫酮(D3T)对大鼠主动脉血管A10细胞GSH和GST的诱导以及D3T增强的细胞防御对HNE介导的毒性的保护作用。A10细胞与D3T孵育后，GSH和GST的诱导均呈明显的浓度依赖性。D3T对细胞GST的诱导也表现出时间依赖性反应。用四甲基偶氮唑盐(3-[4，5-dimethylthiazol-2-yl]-2，5-diphenyltetrazolium)还原试验和扫描电子显微镜检测，D3T可显著降低HNE对A10细胞的细胞毒作用。HNE与A10细胞孵育0.5h和1h后，细胞内GSH显著减少，细胞存活率下降。为进一步证实GSH和GST对HNE诱导的细胞毒性的保护作用，分别用丁硫氨酸亚磺胺(BSO)和柳氮磺吡啶抑制细胞内GSH的合成和GST的活性。BSO耗竭GSH或柳氮磺胺吡啶抑制GST均可显著增强HNE介导的细胞毒作用。此外，BSO与A10细胞共处理可完全阻断D3T介导的GSH诱导，并在很大程度上逆转D3T对HNE诱导的毒性的细胞保护作用。综上所述，本研究表明，D3T可诱导主动脉平滑肌细胞GSH和GST，并且D3T增强的细胞防御对HNE诱导的血管细胞损伤具有明显的保护作用。
4-Hydroxy-2-nonenal (HNE) has been suggested to contribute to the pathogenesis of atherosclerosis. One of the major metabolic transformation pathways of HNE involves conjugation with glutathione (GSH) catalyzed by GSH S-transferase (GST). In this study, we have characterized the induction of GSH and GST by 3H-1,2-dithiole-3-thione (D3T) and the protective effects of the D3T-elevated cellular defenses on HNE-mediated toxicity in rat aortic smooth muscle A10 cells. Incubation of A10 cells with D3T resulted in a marked concentration- dependent induction of both GSH and GST. The induction of cellular GST by D3T also exhibited a time-dependent response. Pretreatment of A10 cells with D3T led to a dramatic decrease of HNE-induced cytotoxicity, as assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) reduction assay and scanning electron microscopy. Incubation of A10 cells with HNE for 0.5 h and 1 h resulted in a significant depletion of cellular GSH, which preceded the decrease of cell viability. To further demonstrate the involvement of GSH and GST in protecting against HNE-induced cytotoxicity, buthionine sulfoximine (BSO) and sulfasalazine were used to inhibit cellular GSH biosynthesis and GST activity, respectively. Either depletion of GSH by BSO or inhibition of GST by sulfasalazine caused great potentiation of HNE-mediated cytotoxicity. Moreover, cotreatment of A10 cells with BSO was found to completely block the D3T-mediated GSH induction and to largely reverse the cytoprotective effects of D3T on HNE-induced toxicity. Taken together, this study demonstrates that D3T can induce both GSH and GST in aortic smooth muscle cells, and that the D3T-augmented cellular defenses afford a marked protection against HNE-induced vascular cell injury.