The role of ANXA5 in DBP-induced oxidative stress through ERK/Nrf2 pathway
The role of ANXA5 in DBP-induced oxidative stress through ERK/Nrf2 pathway
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ANXA5 通过 ERK/Nrf2 通路在 DBP 诱导的氧化应激中的作用
DOI：
10.1016/j.etap.2019.103236
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发表时间：
2019
影响因子：
4.3
通讯作者：
Zhang Wei
中科院分区：
文献类型：
--
作者：
Zhang Lei;Qin Zhiqiang;Li Ran;Wang Shangqian;Wang Wei;Tang Min;Zhang Wei
邻苯二甲酸二丁酯（DBP）因其对男性生殖系统独特的内分泌毒性而引起越来越多的关注。本课题组前期研究表明，抗氧化Nrf 2（nuclear factor erythroid related factor 2）通路在DBP诱导的氧化应激损伤中起重要作用。ANXA 5（膜联蛋白A5），这是高度表达在睾丸Leydig和Sertoli细胞，被发现上调DBP刺激后。用不同浓度的DBP处理小鼠睾丸间质细胞和睾丸支持细胞24 h，观察细胞内活性氧（ROS）的变化（活性氧），MDA（丙二醛），SOD（超氧化物歧化酶）水平和ANXA 5、Nrf 2、NQO 1（NAD（P）H-醌氧化还原酶1）、HO-1（血红素加氧酶1）和ERK/P-ERK蛋白表达（Dihydroethidium）染色、ELISA（酶联免疫吸附试验）和Western blot。首先，重新验证了DBP诱导的氧化应激损伤。然后，我们证实了DBP暴露于睾丸细胞后Nrf 2通路和ANXA 5水平的变化。此外，过表达ANXA 5可以激活Nrf 2/HO-1/NQO 1抗氧化途径，显著减轻DBP诱导的氧化应激。最终，我们证明ANXA 5可以增加ERK磷酸化水平，并且ANXA 5对ERK/Nrf 2通路的激活作用可以被ERK抑制剂逆转。总之，本研究表明ANXA 5可以通过ERK/Nrf 2途径保护睾丸Leydig和Sertoli细胞免受DBP诱导的氧化应激损伤。
Di-N-butylphthalate (DBP) have given rise to more and more attention due to its unique endocrine toxicity to male reproductive system. Our previous studies have demonstrated antioxidative Nrf2 (nuclear factor erythroid related factor 2) pathway play a vital role in DBP induced oxidative stress injury. ANXA5 (annexin A5), which is highly expressed in testicular Leydig and Sertoli cells, was found upregulated after DBP stimulation. Mouse Leydig and Sertoli cells were exposed to different concentration of DBP for 24 h to examine the ROS (Reactive oxygen species), MDA (Malondialdehyde), SOD (superoxide dismutase) level and ANXA5, Nrf2, NQO1 (NAD(P)H-quinone oxidoreductase 1), HO-1 (heme oxygenase 1) and ERK/P-ERK protein expression by DHE (Dihydroethidium) staining, ELISA (enzyme-linked immunosorbent assay) and Western blot respectively. Firstly, the oxidative stress injury induced by DBP was re-validated. Then, we confirmed the change of Nrf2 pathway and ANXA5 level after DBP exposure to testicular cells. Additionally, overexpressed ANXA5 could activate Nrf2/HO-1/NQO1 antioxidant pathway and significantly attenuate DBP-induced oxidative stress. Ultimately, we demonstrated ANXA5 could increase ERK phosphorylated level and the activated role of ANXA5 on ERK/Nrf2 pathway could be reversed by ERK inhibitor. Overall, this study illuminated that ANXA5 could defend testicle Leydig and Sertoli cells against DBP-induced oxidative stress injury through ERK/Nrf2 pathway.