Inhibition of Protein arginine methyltransferase 6 reduces reactive oxygen species production and attenuates aminoglycoside- and cisplatin-induced hair cell death
Inhibition of Protein arginine methyltransferase 6 reduces reactive oxygen species production and attenuates aminoglycoside- and cisplatin-induced hair cell death
复制标题
抑制蛋白质精氨酸甲基转移酶 6 可减少活性氧的产生并减弱氨基糖苷类和顺铂诱导的毛细胞死亡
DOI：
10.7150/thno.37362
复制
发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Li, Huawei
中科院分区：
文献类型：
--
作者：
He, Yingzi;Li, Wen;Li, Huawei
内耳中的毛细胞已被证明对一些有益的药物如氨基糖苷类和顺铂的耳毒性敏感。因此，人们对发现保护毛细胞免受这些耳毒性药物影响的新靶点或化合物非常感兴趣。表观遗传调控与内耳发育密切相关，然而，关于表观遗传调控在耳毒性药物引起的听力损失过程中的作用知之甚少。研究方法：在这项研究中，我们研究了蛋白质精氨酸甲基转移酶6（PRMT 6）在氨基糖苷类和顺铂诱导的毛细胞损失的作用，通过使用EPZ 020411，一种选择性的小分子PRMT 6抑制剂，在体外新生小鼠耳蜗外植体和C57 BL/6小鼠体内。我们还利用HEI-OC 1细胞系来评估PRMT 6敲低对顺铂诱导的耳毒性的抗凋亡作用。使用切割的半胱氨酸蛋白酶-3染色和原位末端标记法鉴定凋亡细胞。DCFH-DA和cellROX绿色染色检测细胞内活性氧（ROS）水平。用JC-1、TMRM和罗丹明123染色法测定线粒体膜电位（Δ Km）。结果：我们发现EPZ 020411显着减轻新霉素和顺铂诱导的细胞凋亡，增加毛细胞存活。此外，EPZ 020411预处理可以减轻体内新霉素和顺铂诱导的听力损失。机制研究表明，抑制PRMT 6可以逆转顺铂损伤后caspase-3和细胞色素c易位的表达增加，线粒体功能障碍，ROS积累增加，以及细胞凋亡的激活。结论：我们的研究结果表明，PRMT 6可能作为一个新的治疗靶点，以防止由氨基糖苷类和顺铂诱导的耳毒性引起的听力损失，通过防止ROS的形成和调节与听力损伤和细胞凋亡有关。
Hair cells in the inner ear have been shown to be susceptible to ototoxicity from some beneficial pharmaceutical drugs, such as aminoglycosides and cisplatin. Thus, there is great interest in discovering new targets or compounds that protect hair cells from these ototoxic drugs. Epigenetic regulation is closely related to inner ear development; however, little is known about epigenetic regulation in the process of ototoxic drugs-induced hearing loss. Methods: In this study, we investigated the role of protein arginine methyltransferase 6 (PRMT6) in aminoglycoside- and cisplatin-induced hair cell loss by using EPZ020411, a selective small molecule PRMT6 inhibitor, in vitro in neonatal mouse cochlear explants and in vivo in C57BL/6 mice. We also took advantage of the HEI-OC1 cell line to evaluate the anti-apoptosis effects of PRMT6 knockdown on cisplatin-induced ototoxicity. Apoptotic cells were identified using cleaved caspase-3 staining and TUNEL assay. The levels of reactive oxygen species (ROS) were evaluated by DCFH-DA and cellROX green staining. The mitochondrial membrane potential (ΔΨm) were determined by JC-1, TMRM, and rhodamine 123 staining. Results: We found that EPZ020411 significantly alleviated neomycin- and cisplatin-induced cell apoptosis and increased hair cell survival. Moreover, pretreatment with EPZ020411 could attenuate neomycin- and cisplatin-induced hearing loss in vivo. Mechanistic studies revealed that inhibition of PRMT6 could reverse the increased expression of caspase-3 and cytochrome c translocation, mitochondrial dysfunction, increased accumulation of ROS, and activation of cell apoptosis after cisplatin injury. Conclusions: Our findings suggested that PRMT6 might serve as a new therapeutic target to prevent hearing loss caused by aminoglycoside- and cisplatin-induced ototoxicity by preventing ROS formation and modulating the mitochondria-related damage and apoptosis.