Wnt/β-Catenin signaling is required for the development and differentiation of cochlear hair cells. Total of 80 natural compounds derived from the FDA-approved Drug Library of Selleck were screened by T-cell factor Reporter Plasmid (TOP)-Flash assay to identify the activation of Wnt/β-Catenin signaling. The mouse cochlear hair cells (HEI-OC1) were treated with cisplatin with or without Guaiacin, and the relative expression of β-Catenin and TRIM33 were detected by qRT-PCR and Western blots. The viability of HEI-OC1 was assayed by MTT method, and mouse cochlear cultures were utilized to detect the Ex vivo survival of cochlear hair cells. Guaiacin was testified to have the most vigorous ability to promote Wnt/β-Catenin signaling among 80 compounds detected, and it can also improve the β-Catenin signaling in mouse cochlear hair cells with up-regulated β-Catenin protein expression, unchanged β-Catenin mRNA expression, and down-regulated TRIM33 expression. Guaiacin increased the viability of HEI-OC1 cells cultured with or without cisplatin, and such a protective effect was also testified in mouse cochlear cultures. Our data indicate that Guaiacin could increase Wnt/β-Catenin signaling by regulating TRIM33/β-Catenin axis, which contributes to the improved survival of cochlear hair cells.

Wnt /β-Catenin信号传导是耳蜗毛细胞发育和分化所必需的。通过T细胞因子报告基因质粒（TOP）-Flash分析法筛选了FDA批准的Selleck药物库中的80种天然化合物，以鉴定Wnt /β-Catenin信号传导的激活。在有或没有愈创霉素的情况下，用顺铂处理小鼠的耳蜗毛细胞（HEI-OC1），并通过qRT-PCR和Western blot检测β-Catenin和TRIM33的相对表达。用MTT法检测HEI-OC1的活力，并利用小鼠耳蜗培养物检测离体耳蜗毛细胞的存活。经验证，愈创霉素在80种化合物中具有最强的Wnt /β-Catenin信号传导能力，还可以改善小鼠耳蜗毛细胞的β-Catenin信号传导，其中β-Catenin蛋白表达上调，而β-Catenin不变。连环蛋白mRNA表达和下调TRIM33表达。愈创霉素增加了在有或没有顺铂的情况下培养的HEI-OC1细胞的活力，在小鼠耳蜗培养物中也证明了这种保护作用。我们的数据表明，愈创霉素可以通过调节TRIM33 /β-Catenin轴来增加Wnt /β-Catenin信号传导，从而有助于改善耳蜗毛细胞的存活率。