He Qiang, Jia Zhanwei, Zhang Ying, Ren Xiumin
E.N.T Department, The Second Hospital of Hebei Medical University, Shijiazhuang, China.
J Cell Mol Med. 2017 Mar;21(3):600-608. doi: 10.1111/jcmm.13005. Epub 2016 Oct 6.
We aimed to investigate the effect of morin hydrate on neural stem cells (NSCs) isolated from mouse inner ear and its potential in protecting neuronal hearing loss. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and bromodeoxyuridine incorporation assays were employed to assess the effect of morin hydrate on the viability and proliferation of in vitro NSC culture. The NSCs were then differentiated into neurons, in which neurosphere formation and differentiation were evaluated, followed by neurite outgrowth and neural excitability measurements in the subsequent in vitro neuronal network. Mechanotransduction of cochlea ex vivo culture and auditory brainstem responses threshold and distortion product optoacoustic emissions amplitude in mouse ototoxicity model were also measured following gentamicin treatment to investigate the protective role of morin hydrate against neuronal hearing loss. Morin hydrate improved viability and proliferation, neurosphere formation and neuronal differentiation of inner ear NSCs, and promoted in vitro neuronal network functions. In both ex vivo and in vivo ototoxicity models, morin hydrate prevented gentamicin-induced neuronal hearing loss. Morin hydrate exhibited potent properties in promoting growth and differentiation of inner ear NSCs into functional neurons and protecting from gentamicin ototoxicity. Our study supports its clinical potential in treating neuronal hearing loss.
我们旨在研究水合桑色素对从小鼠内耳分离的神经干细胞(NSCs)的作用及其在保护神经元性听力损失方面的潜力。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-溴化四氮唑(MTT)和溴脱氧尿苷掺入试验来评估水合桑色素对体外NSC培养物活力和增殖的影响。然后将NSCs分化为神经元,评估神经球的形成和分化,随后在体外神经元网络中测量神经突生长和神经兴奋性。在庆大霉素处理后,还测量了离体耳蜗培养物的机械转导以及小鼠耳毒性模型中的听觉脑干反应阈值和畸变产物耳声发射幅度,以研究水合桑色素对神经元性听力损失的保护作用。水合桑色素改善了内耳NSCs的活力和增殖、神经球形成和神经元分化,并促进了体外神经元网络功能。在离体和体内耳毒性模型中,水合桑色素均预防了庆大霉素诱导的神经元性听力损失。水合桑色素在促进内耳NSCs生长和分化为功能性神经元以及保护免受庆大霉素耳毒性方面表现出强大的特性。我们的研究支持其在治疗神经元性听力损失方面的临床潜力。