MicroRNA-140-5p attenuated oxidative stress in Cisplatin induced acute kidney injury by activating Nrf2/ARE pathway through a Keap1-independent mechanism
MicroRNA-140-5p attenuated oxidative stress in Cisplatin induced acute kidney injury by activating Nrf2/ARE pathway through a Keap1-independent mechanism
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DOI：
10.1016/j.yexcr.2017.09.019
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发表时间：
2017-11-15
影响因子：
3.7
通讯作者：
Zhang, Wen
中科院分区：
文献类型：
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作者：
Liao, Weitang;Fu, Zongjie;Zhang, Wen
氧化应激主要参与急性肾损伤(AKI)的发病机制。最近的研究报道了特定的microRNAs(MiRNAs)对氧化应激的保护作用。因此，我们研究了miR140-5p的水平及其在顺铂诱导的MU发病机制中的作用。建立顺铂诱导的小鼠急性心肌梗死模型。我们发现miR-140-5p在小鼠肾脏中的表达明显增加。生物信息学分析表明，核因子红系2相关因子(Nrf2)是miR-140-5p的潜在靶点，我们证明miR-140-5p不影响Kelch样ECH相关蛋白1(Keap1)的水平，但直接靶向Nrf2基因的3‘-UTR，并通过荧光素酶活性测定和蛋白质印迹证实了其在Nrf2表达调控中的积极作用。此外，与miR140-5p的表达一致，Nrf2的mRNA和蛋白水平以及抗氧化反应元件(ARE)驱动的基因血红素氧合酶-1(HO-1)和NAD(P)H：苯醌氧化还原酶1(NQO1)在小鼠肾脏组织中显著增加。体外研究表明，在HK2细胞中，miR-140-5p的增强表达通过降低ROS水平和增加锰超氧化物歧化酶(MnSOD)的表达而显著减轻氧化应激。同时，miR-140-5p可减少顺铂诱导的HK2细胞乳酸脱氢酶(LDH)漏出，提高细胞活力。然而，转染针对Nrf2的siRNA的HK2细胞取消了miR-140-5p对氧化应激的保护作用。这些结果表明miR-140-5p可能通过靶向Nrf2发挥其抗氧化应激作用。我们的发现表明miR140-5p在Nrf2表达中具有新的转录作用，miR-140-5p通过激活Nrf2依赖的抗氧化途径来保护Nrf2和miR-140-5p免受顺铂诱导的氧化应激，为急性肾损伤提供了潜在的治疗靶点。
Oxidative stress was predominantly involved in the pathogenesis of acute kidney injury (AKI). Recent studies had reported the protective role of specific microRNAs (miRNAs) against oxidative stress. Hence, we investigated the levels of miR140-5p and its functional role in the pathogenesis of Cisplatin induced MU. A mice Cisplatin induced-AKI model was established. We found that miR-140-5p expression was markedly increased in mice kidney. Bioinformatics analysis revealed nuclear factor erythroid 2-related factor (Nrf2) was a potential target of miR-140-5p, We demonstrated that miR-140-5p did not affect Kelch-like ECH-associated protein 1 (Keap1) level but directly targeted the 3'-UTR of Nrf2 mRNA and played a positive role in the regulation of Nrf2 expression which was confirmed by luciferase activity assay and western blot. What was more, consistent with miR140-5p expression, the mRNA and protein levels of Nrf2, as well as antioxidant response element (ARE)-driven genes Heme Oxygenase-1 (HO-1) and NAD(P)H:quinone oxidoreductase 1 (NQO1) were significantly increased in mice kidney tissues. In vitro study, Enforced expression of miR-140-5p in HK2 cells significantly attenuated oxidative stress by decreasing ROS level and increasing the expression of manganese superoxide dismutase (MnSOD). Simultaneously, miR-140-5p decreased lactate dehydrogenase (LDH) leakage and improved cell vitality in HK2 cells under Cisplatin-induced oxidative stress. However, HK2 cells transfected with a siRNA targeting Nrf2 abrogated the protective effects of miR-140-5p against oxidative stress. These results indicated that miR-140-5p might exert its anti-oxidative stress function via targeting Nrf2. Our findings showed the novel transcriptional role of miR140-5p in the expression of Nrf2 and miR-140-5p protected against Cisplatin induced oxidative stress by activating Nrf2-dependent antioxidant pathway, providing a potentially therapeutic target in acute kidney injury.