We aimed to investigate the beneficial effect of Celastrol on inner ear stem cells and potential therapeutic value for hearing loss. The inner ear stem cells were isolated and characterized from utricular sensory epithelium of adult mice. The stemness was evaluated by sphere formation assay. The relative expressions of Atoh1, MAP-2 and Myosin VI were measured by RT-PCR and immunoblotting. The up-regulation of MAP-2 was also analysed with immunofluorescence. The in vitro neuronal excitability was interrogated by calcium oscillation. The electrophysiological property was determined by inward current recorded on patch clamp. Our results demonstrated that Celastrol treatment significantly improved the viability and proliferation of mouse inner ear stem cells, and facilitated sphere formation. Moreover, Celastrol stimulated differentiation of mouse inner ear stem cells to neuronal-like cells and enhanced neural excitability. Celastrol also enhanced neuronal-like cell identity in the inner ear stem cell derived neurons, as well as their electrophysiological function. Most notably, these effects were apparently associated with the upregulation of Atoh1 in response to Celastrol treatment. Celastrol showed beneficial effect on inner ear stem cells and held therapeutic promise against hearing loss.

我们旨在研究Celastrol对内耳干细胞的有益作用以及对听力损失的潜在治疗价值。从成年小鼠的子宫感觉上皮中分离并鉴定了内耳干细胞。通过球形成试验评价茎干。通过RT-PCR和免疫印迹法检测Atoh1，MAP-2和肌球蛋白VI的相对表达。还用免疫荧光分析了MAP-2的上调。钙振荡询问了体外神经元兴奋性。电生理特性由膜片钳上记录的内向电流确定。我们的结果表明，Celastrol处理可显着改善小鼠内耳干细胞的活力和增殖，并促进球的形成。此外，Celastrol刺激小鼠内耳干细胞分化为神经元样细胞并增强神经兴奋性。Celastrol还增强了内耳干细胞衍生神经元中神经元样细胞的身份以及它们的电生理功能。最明显的是，这些作用显然与Celastrol治疗反应中Atoh1的上调有关。Celastrol对内耳干细胞显示出有益的作用，并有望预防听力损失。