miR-26a is Involved in Glycometabolism and Affects Boar Sperm Viability by Targeting PDHX
miR-26a is Involved in Glycometabolism and Affects Boar Sperm Viability by Targeting PDHX
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miR-26a 参与糖代谢并通过靶向 PDHX 影响公猪精子活力。
DOI：
10.3390/cells9010146
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发表时间：
2020-01-01
期刊：
影响因子：
6
通讯作者：
Zeng, Changjun
中科院分区：
文献类型：
--
作者：
Wang, Wencan;Liang, Kai;Zeng, Changjun
miR-26 a与精子代谢相关，可影响精子活力和凋亡。然而，miR-26 a如何影响精子活力在很大程度上仍然未知。我们的前期研究表明PDHX基因被预测为miR-26 a的潜在靶点，其负责丙酮酸氧化脱羧，这被认为是连接糖酵解和氧化磷酸化的关键步骤。在这项研究中，我们首次报道了miR-26 a和PDHX及其在新鲜、冻融和附睾精子中的表达之间的潜在关系。然后，在转染miR-26 a后测定精子活力和存活率。采用RT-qPCR和Western Blot（WB）方法检测PDHX基因在猪精液中的mRNA和蛋白表达水平。结果表明，在精子从附睾头向体、尾转运过程中，PDHX的表达水平显著增加。新鲜精子中PDHX的表达显著高于附睾尾和冻融精子（P < 0.05）。附睾体精子中miR-26 a的表达显著高于头、尾精子（P < 0.05）。此外，在液体储存条件下，miR-26 a模拟物和抑制剂转染猪精子后，分别观察到miR-26 a模拟物和抑制剂处理的精子活力最低和最高（P < 0.05）。转染miR-26 a模拟物和抑制剂24和48 h后，PDHX蛋白水平分别显著低于和高于阴性对照组（P < 0.05）。总之，我们的研究的新颖和诱人的发现提供了合理的证据，即miR-26 a通过PDHX，糖酵解和氧化磷酸化之间的联系，可以调节糖代谢途径，最终影响公猪精子的活力和存活。
miR-26a is associated with sperm metabolism and can affect sperm motility and apoptosis. However, how miR-26a affects sperm motility remains largely unknown. Our previous study indicated that the PDHX gene is predicted to be a potential target of miR-26a, which is responsible for pyruvate oxidative decarboxylation which is considered as a key step for connecting glycolysis with oxidative phosphorylation. In this study, we first reported a potential relationship between miR-26a and PDHX and their expressions in fresh, frozen-thawed, and epididymal boar sperm. Then, sperm viability and survival were determined after transfection of miR-26a. mRNA and protein expression level of PDHX in the liquid-preserved boar sperm after transfection were also determined by RT-qPCR and Western Blot (WB). Our results showed that expression level of PDHX was significantly increased during sperm transit from epididymal caput to corpus and cauda. Similarly, expression of PDHX was significantly higher (P < 0.05) in fresh sperm as compared to epididymal cauda and frozen-thawed sperm. However, the expression of miR-26a in epididymal corpus sperm was significantly higher (P < 0.05) than that of caput and cauda sperm. Furthermore, after transfection of boar sperm with miR-26a mimic and inhibitor under liquid storage, the lowest and highest sperm viability was observed in miR-26a mimic and inhibitor treatment (P < 0.05), respectively. The protein levels of PDHX, after 24 and 48 h of transfection of miR-26a mimics and inhibitor, were notably decreased and increased (P < 0.05), respectively, as compared to negative control (NC) group. In conclusion, the novel and enticing findings of our study provide a reasonable evidence that miR-26a via PDHX, a link between glycolysis and oxidative phosphorylation, could regulate the glycometabolic pathway which eventually affect boar sperm viability and survival.