Overexpression of miR-506-3p Aggravates DBP-Induced Testicular Oxidative Stress in Rats by Downregulating ANXA5 via Nrf2/HO-1 Signaling Pathway.
Overexpression of miR-506-3p Aggravates DBP-Induced Testicular Oxidative Stress in Rats by Downregulating ANXA5 via Nrf2/HO-1 Signaling Pathway.
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miR-506-3p 过表达通过 Nrf2/HO-1 信号通路下调 ANXA5，加重 DBP 诱导的大鼠睾丸氧化应激
DOI：
10.1155/2020/4640605
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发表时间：
2020
影响因子：
--
通讯作者：
Zhang W
中科院分区：
文献类型：
--
作者：
Tang M;Zhang L;Zhu Z;Li R;Wang S;Wang W;Qin Z;Zhang W
背景邻苯二甲酸二丁酯(DBP)是一种独特的内分泌毒性，与影响男性生殖系统的激素紊乱有关，已引起越来越多的关注。然而，DBP致睾丸损伤的机制尚不清楚。本研究旨在通过ANXA5(Annexin A5)/Nrf2/HO-1信号通路，探讨miR-506-3p在DBP诱导的大鼠睾丸氧化应激损伤中的可能分子机制。方法雄性青春期大鼠40只，2周后给予DBP 80 0 mg/kg/d和玉米油1 m L/kg/d灌胃。其中部分大鼠皮下注射2 阿莫尔-506-3P和/或10 重组大鼠ANXA5。通过组织学检查评价睾丸组织的病理形态变化，并评价抗氧化因子。用Western blotting或免疫组织化学方法检测ANXA5、Nrf2及其依赖的抗氧化酶HO-1、NQO1和GST的表达。体外用TM3细胞(间质细胞)检测CCK-8对细胞活性的影响及DBP处理组细胞活性的变化。结果分析了DBP组与正常组大鼠睾丸组织中miRNAs的表达差异，qRT-PCR结果显示，DBP组大鼠睾丸组织中miR-506-3p表达水平较高。DBP处理组大鼠出现严重的炎性细胞浸润，异常生殖细胞增多，生精小管内常有缺失细胞层，导致氧化应激，睾丸功能下降。同时，miR-506-3p的上调加剧了上述变化。此外，miR-506-3p直接与ANXA5结合，过表达miR-506-3p可降低ANXA5的表达，并降低Nrf2/HO-1信号通路的蛋白水平。此外，我们还发现，重组大鼠ANXA5逆转了DBP对miR-506-3p所致大鼠睾丸氧化应激损伤的促进作用。体内实验结果在体外实验中重现。结论miR-506-3p可通过抑制ANXA5的表达，下调Nrf2/HO-1信号通路，加重DBP所致的体内、外睾丸氧化应激损伤，为DBP致睾丸损伤的治疗提供了新的认识。
Background Di-N-butylphthalate (DBP) is a kind of unique endocrine toxicity linked to hormonal disruptions that affects the male reproductive system and has given rise to more and more attention. However, the mechanism of DBP-induced testicular injury remains unclear. Here, the objective of this study was to investigate the potential molecular mechanism of miR-506-3p in DBP-induced rat testicular oxidative stress injury via ANXA5 (Annexin A5)/Nrf2/HO-1 signaling pathway. Methods In vivo, a total of 40 adolescent male rats were treated from 2 weeks with 800 mg/kg/day of DBP in 1 mL/kg corn oil administered daily by oral gavage. Among them, some rats were also injected subcutaneously with 2 nmol agomir-506-3p and/or 10 nmol recombinant rat ANXA5. The pathomorphological changes of testicular tissue were assessed by histological examination, and the antioxidant factors were evaluated. Subsequently, ANXA5, Nrf2, and its dependent antioxidant enzymes, such as HO-1, NQO1, and GST, were detected by Western blotting or immunohistochemical staining. In vitro, TM3 cells (Leydig cells) were used to detect the cell activity by CCK-8 and the transfection in the DBP-treated group. Results Differentially expressed miRNAs between the DBP-treated and normal rats were analyzed, and qRT-PCR showed miR-506-3p was highly expressed in testicular tissues of the DBP-treated rats. DBP-treated rats presented severe inflammatory infiltration, increased abnormal germ cells, and missed cell layers frequently existed in seminiferous tubules, resulted in oxidative stress and decreased testicular function. Meanwhile, upregulation of miR-506-3p aggravated the above changes. In addition, miR-506-3p directly bound to ANXA5, and overexpression of miR-506-3p could reduce the ANXA5 expression and also decrease the protein levels of Nrf2/HO-1 signaling pathway. Additionally, we found that recombinant rat ANXA5 reversed the DBP-treated testicular oxidative stress promoting injury of miR-506-3p in rats. In vivo results were reproduced in in vitro experiments. Conclusions This study provided evidence that miR-506-3p could aggravate the DBP-treated testicular oxidative stress injury in vivo and in vitro by inhibiting ANXA5 expression and downregulating Nrf2/HO-1 signaling pathway, which might provide novel understanding in DBP-induced testicular injury therapy.
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影响因子：
2.9
作者：
He Z;Sun Q;Liang YJ;Chen L;Ge YF;Yun SF;Yao B
通讯作者：
Yao B
影响因子：
4.3
作者：
Foster, PMD;Cattley, RC;Mylchreest, E
通讯作者：
Mylchreest, E
影响因子：
3.8
作者：
Bouter, A.;Carmeille, R.;Brisson, A. R.
通讯作者：
Brisson, A. R.
影响因子：
3.8
作者：
Virtanen, H E;Rajpert-De Meyts, E;Toppari, J
通讯作者：
Toppari, J
影响因子：
4.3
作者：
Wang, Xianliang;Yan, Xu;Yan, Biao
通讯作者：
Yan, Biao