LLY-283在顺铂诱导的内耳毛细胞损伤中的保护作用研究

LLY-283 Antagonizes Cisplatin-induced Damage to Auditory Hair Cells

为了探究蛋白精氨酸甲基转移酶5(protein arginine methyltransferase 5,PRMT5)选择性抑制剂LLY-283对经顺铂诱导的内耳毛细胞损伤作用的影响,本研究首先建立顺铂诱导的HEI-OC1细胞损伤模型,确定合适损伤浓度后,设置对照组、顺铂损伤组及LLY-283预处理实验组(给予40~140μmol/L梯度浓度的LLY-283预处理2 h后,再加入顺铂损伤24 h),通过CCK-8实验检测各组细胞存活率,并采用流式细胞术观察凋亡情况。随后,建立新生鼠基底膜体外培养模型和斑马鱼模型,进一步验证LLY-283在顺铂诱导的毛细胞损伤中的保护作用。结果显示,与顺铂损伤组相比,LLY-283可显著降低顺铂的细胞毒性,抑制顺铂诱导的HEI-OC1细胞凋亡,增加HEI-OC1细胞的存活率;同时,LLY-283能够降低顺铂诱导的小鼠基底膜毛细胞死亡率,并减轻顺铂对斑马鱼侧线毛细胞的损伤作用。实验初步表明,LLY-283可拮抗顺铂的毛细胞毒性并保护听觉毛细胞,有望成为防治顺铂诱发药物性耳聋的潜在药物。

To investigate the effect of protein arginine methyltransferase 5(PRMT5)selective inhibitor LLY-283 on cisplatin-induced inner ear hair cell injury,a cisplatin-induced HEI-OC1 cell injury model was first established.After the appropriate injury concentration was determined,the control group,cisplatin injury group and LLY-283 pretreatment group were set up.For the LLY-283 pretreatment group,the cells were pretreated with gradient concentrations of LLY-283(40~140μmol/L)for 2 h,and then treated with cisplatin for 24 h.The cell viability was detected by CCK-8 assay,and the apoptosis was observed by flow cytometry.Subsequently,an in vitro culture system of neonatal rat basement membrane was established and a zebrafish model was introduced to further verify the protective effect of LLY-283 on cisplatin-induced auditory hair cell damage.The results showed that,compared with the cisplatin injury group,the LLY-283 pretreatment group showed a significant reduction in cytotoxic effects of cisplatin,an increased survival rate of HEI-OC1 cells,and significant inhibition of the apoptosis induced by cisplatin.At the same time,LLY-283 reduced cisplatin-induced mortality of hair cells in the basement membrane of mice and also reduced cisplatin-induced damage to zebrafish lateral line hair cells.The preliminary experiments revealed that LLY-283 could antagonize the toxicity of cisplatin and protect auditory hair cells,indicating that the PRMT5 inhibitor may be a potential drug for the prevention and treatment of cisplatin-induced hearing loss.