Guan Ming, Zhang Yuhua, Huang Qiusheng, He Li, Fang Qiaojun, Zhang Jie, Gao Song, Fang Jia, Ma Yongming, Su Kaiming, Gao Xia
Department of Otolaryngology, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine Hangzhou 310006, China.
Department of Otolaryngology, The Affiliated Hangzhou Hospital of Nanjing Medical University Hangzhou 310006, China.
Am J Transl Res. 2019 Mar 15;11(3):1343-1358. eCollection 2019.
Aging and exposure to noise or ototoxic drugs are major causes of hair cell death leading to human hearing loss, and many agents have been developed to protect hair cells from apoptosis. Fetal bovine serum (FBS) is a fundamental ingredient in the culture medium of hair cell-like House Ear Institute Organ of Corti 1 (HEI-OC-1) cells, but there have been no reports about the function of FBS in HEI-OC-1 cell apoptosis. In this study, we found that FBS deprivation alone significantly increased HEI-OC-1 cell apoptosis in the absence of neomycin exposure and that the presence of FBS significantly inhibited HEI-OC-1 cell apoptosis after neomycin exposure compared to FBS-deprived cells. Further, we found that the protective effect of FBS was dose dependent and more effective than the growth factors B27, N2, EGF, bFGF, IGF-1, and heparan sulfate. We also found that FBS deprivation significantly disrupted the expression level of mitochondrial proteins, increased pro-apoptotic gene expression, decreased the mitochondrial membrane potential, and increased reactive oxygen species accumulation in HEI-OC-1 cells after neomycin exposure. These findings indicate that FBS is involved in maintaining the level of mitochondrial proteins, maintaining the balance of oxidant gene expression, and preventing the accumulation of ROS, and thus FBS maintains normal mitochondrial function and inhibits apoptosis in HEI-OC-1 cells after neomycin exposure.
衰老以及暴露于噪音或耳毒性药物是导致人类听力丧失的毛细胞死亡的主要原因,并且已经开发出许多药物来保护毛细胞免于凋亡。胎牛血清(FBS)是类毛细胞House Ear Institute Corti器官1(HEI-OC-1)细胞培养基中的一种基本成分,但关于FBS在HEI-OC-1细胞凋亡中的功能尚无报道。在本研究中,我们发现单独剥夺FBS在未暴露于新霉素的情况下会显著增加HEI-OC-1细胞凋亡,并且与剥夺FBS的细胞相比,FBS的存在显著抑制了新霉素暴露后HEI-OC-1细胞的凋亡。此外,我们发现FBS的保护作用呈剂量依赖性,并且比生长因子B27、N2、表皮生长因子(EGF)、碱性成纤维细胞生长因子(bFGF)、胰岛素样生长因子-1(IGF-1)和硫酸乙酰肝素更有效。我们还发现,剥夺FBS会显著破坏新霉素暴露后HEI-OC-1细胞中线粒体蛋白的表达水平,增加促凋亡基因的表达,降低线粒体膜电位,并增加活性氧的积累。这些发现表明,FBS参与维持线粒体蛋白水平、维持氧化基因表达的平衡以及防止活性氧的积累,因此FBS维持正常的线粒体功能并抑制新霉素暴露后HEI-OC-1细胞的凋亡。