"KLF9 regulates PRDX6 expression in hyperglycemia-aggravated bupivacaine neurotoxicity
"KLF9 regulates PRDX6 expression in hyperglycemia-aggravated bupivacaine neurotoxicity
复制标题
“KLF9 在高血糖加重的布比卡因神经毒性中调节 PRDX6 的表达
DOI：
10.1007/s11010-021-04059-8
复制
发表时间：
2021
影响因子：
4.3
通讯作者：
李乐
中科院分区：
文献类型：
--
作者：
李慧;翁雅倩;赖露颖;雷洪伊;徐世元;张洋;李乐
背景局麻药的神经毒性是一种潜在的危及生命的药物，尤其是对糖尿病等基础疾病患者。麻醉剂布比卡因（Bup）已被报道诱导由活性氧（ROS）介导的神经毒性，高血糖会加重这种毒性。Krüppel样因子9（KLF 9）是一种轴突生长抑制转录因子，在神经元成熟和促进氧化应激中起关键作用。本研究旨在探讨KLF 9是否以及如何调节高血糖条件下LA神经毒性相关的ROS水平。在正常或高糖（HG）培养条件下培养KLF 9缺陷细胞和正常细胞，然后暴露于Bup。检测细胞活力、细胞内和线粒体ROS以及线粒体膜电位（Δ Δ μ m）以检查KLF 9的作用。此后，KLF 9缺陷和正常细胞进行预处理与小干扰RNA靶向peroxiredoxin 6（siRNA-Prdx 6），以确定是否PRDX 6是目标蛋白在HG加重Bup neurotoxicity.ResultsThe mRNA和蛋白水平的KLF 9增加后Bup和高血糖治疗。此外，与Sh-Ctrl相比，Sh-Klf 9处理后细胞存活率和线粒体功能显著改善，ROS产生减少。此外，PRDX 6的表达在高血糖培养中被Bup抑制，并且在Sh-Klf 9组中被上调。结论高血糖可通过上调KLF 9的表达，抑制抗氧化剂PRDX 6的表达，导致线粒体功能紊乱，ROS爆发，细胞死亡，从而加重Bup的神经毒性。因此，了解这一机制可能为预防和治疗LAs诱导的神经毒性，特别是糖尿病患者的神经毒性提供有价值的见解。
BackgroundNeurotoxicity induced by local anesthetics (LAs) is potentially life threatening, especially for patients with underlying diseases like diabetes. The anesthetic bupivacaine (Bup) has been reported to induce neurotoxicity mediated by reactive oxygen species (ROS), which is aggravated by hyperglycemia. Krüppel-like factor 9 (KLF9), an axon growth-suppressing transcription factor, plays a key role in neuronal maturation and promotes oxidative stress. This study was designed to investigate whether and how KLF9 regulates ROS levels related to LA neurotoxicity under hyperglycemic conditions.MethodsKlf9/GFP ShRNA (LV Sh-Klf9) was used to achieve stableKlf9knockdown in the SH-SY5Y cell line. KLF9-deficient and normal cells were cultured under normal or high-glucose (HG) culture conditions and then exposed to Bup. Cell viability, intracellular and mitochondrial ROS, and mitochondrial membrane potential (ΔΨm) were detected to examine the role of KLF9. Thereafter, KLF9-deficient and normal cells were pretreated with small-interfering RNA targeting peroxiredoxin 6 (siRNA-Prdx6) to determine if PRDX6 was the target protein in HG-aggravated Bup neurotoxicity.ResultsThe mRNA and protein levels of KLF9 were increased after Bup and hyperglycemia treatment. In addition, cell survival and mitochondrial function were significantly improved, and ROS production was decreased after Sh-Klf9treatment compared with Sh-Ctrl. Furthermore, the expression of PRDX6 was suppressed by Bup in hyperglycemic cultures and was upregulated in theSh-Klf9group. Moreover, the protection provided by KLF9 deficiency for cell survival, the increase in ROS production in cells and mitochondria, and the disruption of mitochondrial function were abolished byPrdx6knockdown.ConclusionsThe results of this study demonstrated that hyperglycemia aggravated Bup neurotoxicity by upregulating KLF9 expression, which repressed the antioxidant PRDX6 and led to mitochondrial dysfunction, ROS burst, and cell death. Understanding this mechanism may, thus, offer valuable insights for the prevention and treatment of neurotoxicity induced by LAs, especially in diabetic patients.