N-Acetylcysteine Prevents Nitric Oxide-Induced Chondrocyte Apoptosis and Cartilage Degeneration in an Experimental Model of Osteoarthritis
N-Acetylcysteine Prevents Nitric Oxide-Induced Chondrocyte Apoptosis and Cartilage Degeneration in an Experimental Model of Osteoarthritis
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DOI：
10.1002/jor.20976
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发表时间：
2010-02-01
影响因子：
2.8
通讯作者：
Kubo, Toshikazu
中科院分区：
文献类型：
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作者：
Nakagawa, Shuji;Arai, Yuji;Kubo, Toshikazu
我们研究了谷胱甘肽前体 N-乙酰半胱氨酸 (NAC) 是否可以保护兔关节软骨细胞免受一氧化氮 (NO) 诱导的细胞凋亡，并在大鼠骨关节炎 (OA) 实验模型中防止软骨破坏。用不同浓度的 NAC (0-2 mM) 处理分离的软骨细胞。 0.75 mM 硝普钠 (SNP) 脱水物诱导细胞凋亡，产生 NO。通过MTT测定评估细胞活力，通过Hoechst 33342和TUNEL染色评估细胞凋亡。测量细胞内活性氧（ROS）和谷胱甘肽水平，并通过蛋白质印迹法测定p53和caspase-3的表达。为了确定关节内注射 NAC 是否可以防止体内软骨破坏，对 OA 模型的软骨样本进行了 H&E、Safranin 0 和 TUNEL 染色。 NAC 可防止 NO 诱导的细胞凋亡、ROS 过量产生、p53 上调和 caspase-3 激活。 NAC的保护作用被谷胱甘肽合成酶抑制剂丁硫氨酸亚砜亚胺显着阻断，表明NAC的细胞凋亡预防活性是由谷胱甘肽介导的。使用实验诱导的 CA 大鼠模型，我们发现 NAC 还可以显着防止体内软骨破坏和软骨细胞凋亡。这些结果表明，NAC在体外通过谷胱甘肽抑制NO诱导的软骨细胞凋亡，在体内抑制软骨细胞凋亡和关节软骨退变。 (C) 2009 年骨科研究学会。出版者：Wiley periodicals, Inc. J Orthop Res 28:156-163, 2010
We investigated whether N-acetyleysteine (NAC), a precursor of glutathione, could protect rabbit articular chondrocytes against nitric oxide (NO)-induced apoptosis and could prevent cartilage destruction in an experimental model of osteoarthritis (OA) in rats. Isolated chondrocytes were treated with various concentrations of NAC (0-2 mM). Apoptosis was induced by 0.75 mM sodium nitroprusside (SNP) dehydrate, which produces NO. Cell viability was assessed by MTT assay, while apoptosis was evaluated by Hoechst 33342 and TUNEL staining. Intracellular reactive oxygen species (ROS) and glutathione levels were measured, and expression of p53 and caspase-3 were determined by Western blotting. To determine whether intraarticular injection of NAC prevents cartilage destruction in vivo, cartilage samples of an OA model were subjected to H&E, Safranin 0, and TUNEL staining. NAC prevented NO-induced apoptosis, ROS overproduction, p53 up-regulation, and caspase-3 activation. The protective effects of NAC were significantly blocked by buthionine sulfoximine, a glutathione synthetase inhibitor, indicating that the apoptosis-preventing activity of NAC was mediated by glutathione. Using a rat model of experimentally induced CA, we found that NAC also significantly prevented cartilage destruction and chondrocyte apoptosis in vivo. These results indicate that NAC inhibits NO-induced apoptosis of chondrocytes through glutathione in vitro, and inhibits chondrocyte apoptosis and articular cartilage degeneration in vivo. (C) 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:156-163, 2010