PKCδ contributes to oxidative stress-induced apoptosis in porcine ovarian granulosa cells via activating JNK
PKCδ contributes to oxidative stress-induced apoptosis in porcine ovarian granulosa cells via activating JNK
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PKC δ 通过激活 JNK 促进氧化应激诱导的猪卵巢颗粒细胞凋亡
DOI：
10.1016/j.theriogenology.2019.03.023
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发表时间：
2019-06-01
期刊：
影响因子：
2.8
通讯作者：
Liu, Honglin
中科院分区：
文献类型：
--
作者：
Liu, Shuo;Shen, Ming;Liu, Honglin
氧化应激诱导的颗粒细胞凋亡被认为是导致卵泡闭锁的重要原因。我们之前的工作表明，c-Jun n末端激酶（也称为JNK）可能促进氧化应激过程中GCs的凋亡。本研究的目的是研究氧化应激过程中jnk介导的GCs凋亡所需的上游信号。由于PKC δ和ASK1被认为在某些类型的细胞中调节JNK活性，我们假设PKC δ和ASK1可能有助于氧化刺激下GCs中JNK依赖性的凋亡。为了验证这一假设，从健康卵泡中获得的猪GCs分别用H2O2单独处理，或与PKC δ和JNK抑制剂一起处理，然后收集细胞活力测定，TUNEL染色，免疫沉淀，western blotting或体外JNK活性检测。目前的研究结果表明，h2o2处理的猪GCs细胞活力丧失、DNA断裂、形态收缩和核凝聚与JNK活化增强有关。虽然ASK1被认为是JNK的激活剂，但我们没有发现ASK1在氧化应激中JNK诱导的GCs凋亡中的明确作用。进一步的研究表明，h2o2介导的PKC δ激活是猪GCs凋亡的必要条件。PKC δ对猪GCs的促凋亡作用可能是通过激活线粒体途径实现的。重要的是，我们发现p-PKC δ在h2o2处理的猪GCs中充当JNK的上游激活剂。然而，JNK对PKC δ活性没有调节作用。综上所述，我们的研究结果提供了一个氧化应激过程中涉及PKC δ /JNK/线粒体凋亡轴激活的GCs凋亡新模型。(C) 2019由爱思唯尔公司出版。
Oxidative stress-induced apoptosis of granulosa cells (GCs) is believed to be an important cause of follicular atresia. Our previous work showed that the c-Jun N-terminal kinase (also known as JNK) might promote apoptosis in GCs during oxidative stress. The aim of this study was to investigate the upstream signaling required for JNK-mediated GCs apoptosis during oxidative stress. Since PKC delta, and ASK1 have been suggested to regulate JNK activity in some types of cells, we hypothesized that PKC delta and ASK1 might contribute to JNK-dependent apoptosis in GCs suffering oxidative stimulation. To test this assumption, porcine GCs obtained from healthy follicles were treated with H2O2 alone, or together with inhibitors against PKC delta and JNK, and then collected for cell viability assay, TUNEL staining, immunoprecipitation, western blotting, or JNK activity detection in vitro. The current results showed that the cell viability loss, DNA fragmentation, morphological shrinkage, and nuclear condensation in H2O2-treated porcine GCs was correlated with enhanced activation of JNK. Although ASK1 was supposed to be a JNK activator, we found no definite role of ASK1 in JNK-induced GCs apoptosis during oxidative stress. Further investigations revealed that H2O2-mediated PKC delta activation was required for the apoptotic death of porcine GCs. Particularly, the pro-apoptotic effects of PKC delta on porcine GCs might be achieved by activating the mitochondrial pathway. Importantly, we found that p-PKC delta acts as an upstream activator of JNK in H2O2-treated porcine GCs. However, JNK has no regulatory effect on PKC delta activity. Taken together, our findings provided a novel model of GCs apoptosis involving the activation of PKC delta/JNK/mitochondrial apoptosis axis during oxidative stress. (C) 2019 Published by Elsevier Inc.