细胞凋亡
免疫印迹
衰老
耳蜗
流式细胞术
化学
污渍
男科
免疫荧光
内耳
细胞生物学
分子生物学
生物
免疫学
医学
解剖
生物化学
抗体
基因
作者
Ziyi Feng,Xuerui Li,Long Chen,Ying Zhou,Yue-Chen Chang,Ketao Ma,Jun‐Qiang Si,Li Li
出处
期刊:Chinese journal of applied physiology
[Chinese Medical Association]
日期:2020-09-01
卷期号:36 (5): 385-389
标识
DOI:10.12047/j.cjap.5959.2020.082
摘要
Objective: Primary cultured cochlear stria vascularis endothelial cells (ECs) of guinea pig were used to investigate the expression changes of TMEM16A and its effect on apoptosis and senescence of ECs in the cochlear stria vascularis. Methods: Primary cultured ECs in the cochlear stria vascularis were used to establish aging models according to CCK-8 and SA-β-galactosidase. Senescent cells were randomly divided into senescent group (P12), DMSO group (P12+DMSO), T16Ainh-A01 group (P12+T16Ainh-A01). Immunofluorescence and Western blot were used to detect the expression of TMEM16A in ECs. Flow cytometry was used to detect the apoptotic rate. Western blot was used to detect the protein expressions of Bax, Bcl-2 and cleaved casepase-3 in each group. Results: The positive rate of primary cultured cochlear stria vascularis ECs was above 95%, and the 12th generation cochlear stria vascularis ECs were determined as the senescence group, and the expression of TMEM16A in protein and fluorescence was increased (P<0.05). After intervention with T16Ainh-A01 for 24 h, the protein expressions of Bax and cleaved casepase-3 were down-regulated (P<0.01), the protein expression of Bcl-2 was increased (P<0.05), the apoptotic rate and the positive rate of SA-β-gal were down-regulated (P<0.01). Conclusion: It was found that apoptosis and TMEM16A expression were increased in cochlear stria vascularis senescent ECs, TMEM16A specific blocker T16Ainh-A01 could reduce the apoptosis and senescence in ECs of the cochlear stria vascularis. These results suggest that TMEM16A may participate in apoptosis and senescence of ECs in the cochlear stria vascularis.
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