Evaluating the Protective Effects and Mechanisms of Diallyl Disulfide on Interlukin-1β-Induced Oxidative Stress and Mitochondrial Apoptotic Signaling Pathways in Cultured Chondrocytes
Evaluating the Protective Effects and Mechanisms of Diallyl Disulfide on Interlukin-1β-Induced Oxidative Stress and Mitochondrial Apoptotic Signaling Pathways in Cultured Chondrocytes
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DOI：
10.1002/jcb.25907
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发表时间：
2017-07-01
影响因子：
4
通讯作者：
Sharifi, Ali M.
中科院分区：
文献类型：
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作者：
Hosseinzadeh, Azam;Jafari, Davood;Sharifi, Ali M.
在 C28I2 人软骨细胞中研究了 DADS 对 IL-1 β 介导的氧化应激和线粒体凋亡的保护作用和机制。在不同时间（2、4、8、16和24小时）评估不同浓度的DADS（1、5、10、25、50和100μM）对C28I2细胞活力的影响，通过MTT测定获得药物的非细胞毒性浓度。评估了无毒浓度的 DADS 对 C28I2 中 IL-1β 实验诱导的氧化应激和细胞凋亡的保护作用。检测DADS对IL-1β诱导的细胞内ROS产生和脂质过氧化的影响，并通过Western blotting分析Nrf2、Bax、Bcl-2、caspase-3、总JNK和磷酸化JNK以及P38 MAPK的蛋白表达。通过逆转录聚合酶链式反应评估解毒II相/抗氧化酶包括血红素加氧酶-1、NAD(P)H奎宁氧化还原酶、谷胱甘肽S-转移酶-P1、过氧化氢酶、超氧化物歧化酶-1、谷胱甘肽过氧化物酶-1、-3、-4的mRNA表达。 1、5、10和25μM浓度的DADS在24小时后没有细胞毒性作用。 DADS 预处理显着增加 Nrf2 核转位以及解毒 II 相/抗氧化酶的基因表达，并减少 IL-1b 诱导的 ROS 升高、脂质过氧化、Bax/Bcl-2 比率、caspase-3 激活以及 JNK 和 P38 磷酸化。 DADS 可以显着减少 IL-1b 诱导的氧化应激和随之而来的线粒体凋亡，这是骨关节炎实验模型中软骨细胞死亡的主要机制。它可被视为在临床环境中保护 OA 引起的软骨损伤的天然产物。 (C) 2017 年 Wiley 期刊公司。
The protective effects and mechanisms of DADS on IL-1 beta-mediated oxidative stress and mitochondrial apoptosis were investigated in C28I2 human chondrocytes. The effect of various concentrations of DADS (1, 5 10, 25, 50, and 100 mu M) on C28I2 cell viability was evaluated in different times (2, 4, 8, 16, and 24 h) to obtain the non-cytotoxic concentrations of drug by MTT-assay. The protective effect of non-toxic concentrations of DADS on experimentally induced oxidative stress and apoptosis by IL-1 beta in C28I2 was evaluated. The effects of DADS on IL-1 beta-induced intracellular ROS production and lipid peroxidation were detected and the proteins expression of Nrf2, Bax, Bcl-2, caspase-3, total and phosphorylated JNK, and P38 MAPKs were analyzed by Western blotting. The mRNA expression of detoxifying phase II/antioxidant enzymes including heme oxygenase-1, NAD(P) H quinine oxidoreductase, glutathione S-transferase-P1, catalase, superoxide dismutase-1, glutathione peroxidase-1, -3, -4 were evaluated by reverse transcriptionpolymerase chain reaction. DADS in 1, 5, 10, and 25 mu M concentrations had no cytotoxic effect after 24 h. Pretreatment with DADS remarkably increased Nrf2 nuclear translocation as well as the genes expression of detoxifying phase II/antioxidant enzymes and reduced IL-1b-induced elevation of ROS, lipid peroxidation, Bax/Bcl-2 ratio, caspase-3 activation, and JNK and P38 phosphorylation. DADS could considerably reduce IL-1b-induced oxidative stress and consequent mitochondrial apoptosis, as the major mechanisms of chondrocyte cell death in an experimental model of osteoarthritis. It may be considered as natural product in protecting OA-induced cartilage damage in clinical setting. (C) 2017 Wiley Periodicals, Inc.