Inhibition of Rho Kinase Induces Antioxidative Molecules and Suppresses Reactive Oxidative Species in Trabecular Meshwork Cells.
Inhibition of Rho Kinase Induces Antioxidative Molecules and Suppresses Reactive Oxidative Species in Trabecular Meshwork Cells.
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DOI：
10.1155/2017/7598140
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发表时间：
2017
影响因子：
1.9
通讯作者：
Tanihara H
中科院分区：
文献类型：
--
作者：
Fujimoto T;Inoue T;Ohira S;Awai-Kasaoka N;Kameda T;Inoue-Mochita M;Tanihara H
探讨rho激酶抑制剂对小梁网（TM）细胞氧化应激的影响。从食蟹猴眼中分离TM细胞。Y-27632和menadione分别抑制rho激酶和诱导活性氧（ROS）的产生。采用食蟹猴阵列和12613个探针进行DNA微阵列分析，采用基因本体分析对影响基因进行分类。通过实时RT-PCR检测靶基因mRNA表达水平。用对活性氧敏感的荧光试剂检测细胞内氧化应激。采用WST-8法测定细胞活力。基因本体分析显示抗氧化活性相关基因上调，Y-27632处理30分钟后，实时RT-PCR证实过氧化氢酶上调。甲萘醌增加了ROS的产生，Y-27632预处理部分抑制了ROS的产生。在较低剂量的甲萘酮下，Y-27632刺激了TM细胞，与对照细胞相比，甲萘酮处理后，Y-27632显著提高了TM细胞的活力。通过微阵列分析，Y-27632被证明可以上调过氧化氢酶等抗氧化基因，并部分减少甲萘醌引起的氧化应激引起的ROS产生和细胞死亡。
To investigate the effect of rho kinase inhibitors on oxidative stress in trabecular meshwork (TM) cells. TM cells were isolated from the eyes of cynomolgus monkeys. Y-27632 and menadione were used to inhibit rho kinase and induce production of reactive oxygen species (ROS), respectively. The cynomolgus monkey array and 12,613 probes were used in DNA microarray analysis, and the affected genes were categorized using gene ontology analysis. The mRNA levels of the target genes were confirmed by real-time RT-PCR. Intracellular oxidative stress was detected using a fluorescent reagent sensitive to ROS. Cell viability was assessed by the WST-8 assay. Gene ontology analysis revealed upregulation of genes involved in antioxidant activity, and upregulation of catalase was confirmed by real-time RT-PCR after 30 min treatment with Y-27632. Production of ROS was increased by menadione, and the effect was partly suppressed by pretreatment with Y-27632. At a lower dose of menadione, Y-27632 stimulated TM cells and significantly increased their viability following menadione treatment compared to control cells. Using microarray analysis, Y-27632 was shown to upregulate antioxidative genes including catalase and partially reduce ROS production and cell death by oxidative stress caused by menadione.
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影响因子：
3.7
作者：
Tanito M;Kaidzu S;Takai Y;Ohira A
通讯作者：
Ohira A
影响因子：
5.9
作者：
Izzotti, A;Saccà, SC;De Flora, S
通讯作者：
De Flora, S
影响因子：
20.3
作者：
Tang, Yixin;Scheef, Elizabeth A.;Sheibani, Nader
通讯作者：
Sheibani, Nader
影响因子：
9
作者：
通讯作者：
--
影响因子：
3.7
作者：
Inoue-Mochita M;Inoue T;Fujimoto T;Kameda T;Awai-Kasaoka N;Ohtsu N;Kimoto K;Tanihara H
通讯作者：
Tanihara H