L-carnitine alleviates oxidative stress-related damage via MAPK signaling in human lens epithelial cells exposed to H2O2
L-carnitine alleviates oxidative stress-related damage via MAPK signaling in human lens epithelial cells exposed to H2O2
复制标题
左旋肉碱通过暴露于 H2O2 的人晶状体上皮细胞中的 MAPK 信号传导减轻氧化应激相关损伤
DOI：
10.3892/ijmm.2019.4283
复制
发表时间：
2019-10-01
影响因子：
5.4
通讯作者：
Yuan, Xiaoyong
中科院分区：
文献类型：
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作者：
Li, Xiaoxia;Meng, Fanlan;Yuan, Xiaoyong
L-肉碱（LC）是众所周知的抗氧化性能。本研究旨在探讨LC对人透镜上皮细胞（HLECs）的影响及其在白内障发生中的调控机制。HLE B-3细胞用过氧化氢（H2 O2）培养，并用或不用LC预处理。细胞计数试剂盒-8测定用于测定细胞活力。采用活性氧（ROS）测定试剂盒检测H2 O2和LC诱导的细胞ROS产生。采用逆转录-定量PCR和western blot检测氧化损伤标志物和抗氧化酶的表达水平。值得注意的是，ROS过量生产后，暴露于H2 O2，而LC补充显着降低ROS水平，通过激活的抗氧化酶叉头盒O 1，过氧化物酶4和过氧化氢酶。此外，LC还抑制了炎症相关基因[白细胞介素（IL）1、IL 6、IL 8和环氧合酶-2]和凋亡相关基因（caspase-3）的表达。相反，LC促进增殖细胞核抗原，细胞周期蛋白依赖性激酶（CDK）2和CDK 4的表达，这可能会增加H2 O2孵育的HLEC的增殖。此外，当活性氧积累时，发生上皮-间充质转化，而H2 O2对AQP 1和波形蛋白表达的影响在补充LC后被逆转。值得注意的是，这项研究表明，LC恢复氧化剂/抗氧化剂的平衡，并通过丝裂原活化蛋白激酶信号通路保护细胞免受损伤。总之，LC可以起到抑制氧化损伤的保护作用，因此可以被认为是治疗白内障的潜在治疗剂。
L-carnitine (LC) is well known for its antioxidative properties. The present study aimed to evaluate the effects of LC on human lens epithelial cells (HLECs) and to analyze its regulatory mechanism in cataractogenesis. HLE B-3 cells were cultured with hydrogen peroxide (H2O2) and were pretreated with or without LC. The Cell Counting kit-8 assay was used to determine cell viability. Reactive oxygen species (ROS) assay kit was used to measure the cellular ROS production induced by H2O2 and LC. In addition, reverse transcription-quantitative PCR and western blot analysis were performed to detect the expression levels of oxidative damage markers and antioxidant enzymes. Notably, ROS overproduction was observed upon exposure to H2O2, whereas LC supplementation markedly decreased ROS levels through activation of the antioxidant enzymes forkhead box O1, peroxiredoxin 4 and catalase. Furthermore, LC suppressed the expression of apoptosis-associated genes (caspase-3) and inflammation-associated genes [interleukin (IL)1, IL6, IL8 and cyclooxygenase-2]. Conversely, LC promoted proliferating cell nuclear antigen, cyclin-dependent kinase (CDK)2 and CDK4 expression, which may increase proliferation of HLECs that were incubated with H2O2. In addition, epithelial-mesenchymal transition occurred upon ROS accumulation, whereas the effects of H2O2 on AQP1 and vimentin expression were reversed upon LC supplementation. Notably, this study revealed that LC restored the oxidant/antioxidant balance and protected against cell damage through the mitogen-activated protein kinase signaling pathway. In conclusion, LC may serve a protective role in curbing oxidative damage and therefore may be considered a potential therapeutic agent for the treatment of cataracts.