LLY-283在顺铂诱导的内耳毛细胞损伤中的保护作用研究

张悦, 韩京宏, 聂国辉

生命科学研究 ›› 2023, Vol. 27 ›› Issue (4) : 298 -304.

细胞与生理学

LLY-283在顺铂诱导的内耳毛细胞损伤中的保护作用研究

张悦, 韩京宏, 聂国辉

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

ZHANG Yue, HAN Jinghong, NIE Guohui

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摘要

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40~140 μmol/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。

Abstract

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 in-jury 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 (40~140 μmol/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-in-duced 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.