Adenoviral-mediated overexpression of catalase inhibits endothelial cell proliferation.
Adenoviral-mediated overexpression of catalase inhibits endothelial cell proliferation.
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DOI：
10.1152/ajpheart.00358.2001
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发表时间：
2002-12
期刊：
影响因子：
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通讯作者：
M. Zanetti;Z. Katusic;T. O’Brien
中科院分区：
文献类型：
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作者：
M. Zanetti;Z. Katusic;T. O’Brien
尽管过氧化氢(H(2)O(2))可诱导血管平滑肌细胞增殖，但其对内皮细胞增殖的作用尚不清楚。我们的目的是通过过表达过氧化氢酶来研究过氧化氢在内皮细胞增殖中的作用。将编码β-半乳糖苷酶(Adbetagal)或过氧化氢酶(AdCat)的腺病毒载体转导人主动脉内皮细胞，或单独暴露于稀释液中(对照组)。经β-半乳糖苷酶染色和Western分析证实转基因表达，并显著提高了AdCat转导细胞的酶活性。与对照组和Adbetagal转导细胞相比，过表达过氧化氢酶使AdCat细胞的DNA合成减少(分别为536.8+/-31比1,875.1+/-132.9比1,347.5+/-93.7DPM/孔；P<；0.05比对照和Adbetagal)。AdCat(感染复数=50)转导6天后，细胞数显著减少(AdCat：对照组细胞数的38+/-1.8%，P<；0.05；Adbetagal组细胞数的45+/-2%，P<；0.05)。用10 mmoL/L的过氧化氢酶抑制剂氨基三氮唑孵育可阻断这一作用。AdCat转导后2d和4d，细胞凋亡数分别增加1倍和3倍。外源性低浓度H(2)O(2)(50微米)显著促进细胞增殖，高浓度则抑制细胞增殖。这些结果表明，H(2O)O(2)是内皮细胞增殖的重要调节因子。
Although hydrogen peroxide (H(2)O(2)) induces proliferation of vascular smooth muscle cells, its role in endothelial cell proliferation is unclear. Our aim was to study the role of hydrogen peroxide in endothelial cell proliferation by overexpressing catalase. Human aortic endothelial cells were transduced with adenoviral vectors encoding beta-galactosidase (Adbetagal) or catalase (AdCat) or were exposed to diluent alone (control). Transgene expression was demonstrated by beta-galactosidase staining, Western analysis, and significantly increased enzyme activity in AdCat-transduced cells. Overexpression of catalase decreased DNA synthesis in AdCat compared with control and Adbetagal-transduced cells (536.8 +/- 31 vs. 1,875.1 +/- 132.9 vs. 1,347.5 +/- 93.7 dpm/well, respectively; P < 0.05 vs. control and Adbetagal). Six days after transduction with AdCat (multiplicity of infection = 50), cell numbers were significantly reduced (AdCat: 38 +/- 1.8% of cell counts in control, P < 0.05; and 45 +/- 2% of cell count in Adbetagal, P < 0.05). Incubation with aminotriazole 10 mmol/l, an inhibitor of catalase, prevented this effect. The number of apoptotic cells was increased one- and threefold 2 and 4 days, respectively, after transduction with AdCat. Exogenous administration of low concentrations of H(2)O(2) (50 microM) significantly increased cell proliferation, whereas it was inhibited by higher concentrations. These results suggest that H(2)O(2) is an important modulator of endothelial cell proliferation.