miR-26a-5p 调控SLC26A4挽救听力减退在耳聋中的作用

丛林海; 汤勇; 殷家志; 邓苙; 高慧芳; 方红丽; 李昕

doi:10.12259/j.issn.2095-610X.S20230603

miR-26a-5p 调控SLC26A4挽救听力减退在耳聋中的作用

doi: 10.12259/j.issn.2095-610X.S20230603

丛林海^1,,
汤勇^1, ,,
殷家志¹,
邓苙²,
高慧芳²,
方红丽²,
李昕³

1.
昆明医科大学第一附属医院耳鼻喉科
2.
健康管理中心
3.
检验科，云南昆明　650032

基金项目: 云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目（202001AY070001-211）

详细信息

作者简介:
丛林海（1972～），男，黑龙江伊春人，医学博士，主任医师，主要从事耳鼻咽喉科临床工作

通讯作者:
汤勇， E-mail：yongtang0929@sina.com

中图分类号: R764.431
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出版历程
- 收稿日期: 2023-03-11
- 网络出版日期: 2023-06-16
- 刊出日期: 2023-06-25

Effect of MiR-26a-5p Regulating SLC26A4 to Ameliorate Hearing Loss in Deafness

1.
Dept. of Otolaryngology
2.
Health Administration Center
3.
Clinical Laboratory，The 1st Affiliated Hospital of Kunming Medical University，Kunming Yunnan 650032，China

摘要

摘要: 目的检测miR-26a-5p和SLC26A4在耳聋小鼠和听力损失小鼠中的表达变化，以此研究miR-26a-5p 调控SLC26A4挽救听力减退的作用。方法通过构建小鼠听力损失模型和小鼠耳聋模型，使用听性脑干反应测试检测小鼠右耳的听力来鉴定模型的构建是否成功。使用qPCR实验检测miR-26a-5p和SLC26A4在小鼠听力损失和耳聋后的相对表达量，通过TargetScan网站预测miR-26a-5p靶向结合SLC26A4的具体位点，对小鼠注射miR-26a-5p和SLC26A4的干扰和过表达载体，继续检测miR-26a-5p和SLC26A4的表达变化。结果在小鼠听力损失和耳聋后，miR-26a-5p表达显著降低（P < 0.05），而SLC26A4表达明显升高（P < 0.05），在小鼠注射antagomir后，miR-26a-5p明显下调（P = 0.047），同时SLC26A4显著上调（P = 0.014），同时小鼠听力阈值明显降低（P < 0.05），而在注射miR-26a-5p agomir后结果恰恰相反。结论 miR-26a-5p可以通过SLC26A4从而降低听力减退以此抑制耳聋的发生。
- 听力减退 /
- 耳聋 /
- miR-26a-5p /
- SLC26A4
Abstract: Objective To explore the effect of miR-26a-5p in regulating SLC26A4 to amiliorate hearing loss, and to detect the expression changes of miR-26a-5p and SLC26A4 in deaf mice and hearing loss mice. Methods A mouse hearing loss model and a mouse deafness model were constructed, and the auditory brainstem response test was used to detect the hearing in the right ear of mice to determine whether the models were successfully constructed. qPCR test was used to detect the relative expression of miR-26a-5p and SLC26A4 after hearing loss and deafness in mice were established. The specific site of miR-26a-5p targeting binding to SLC26A4 was predicted at TargetScan website. The interference and overexpression vector of miR-26a-5p and SLC26A4 were injected, and the expression changes of miR-26a-5p and SLC26A4 were detected. Results After hearing loss and deafness were established, the expression of miR-26a-5p was significantly decreased （P < 0.05）, while the expression of SLC26A4 was significantly increased （P < 0.05）. After antagomir injection, miR-26a-5p was significantly down-regulated （P = 0.047）, while SLC26A4 was significantly up-regulated （P = 0.014）; and the hearing threshold of mice decreased significantly （P < 0.05）, but the result was just the opposite after injection of miR-26a-5p agomir. Conclusion miR-26a-5p can ameliorate hearing loss through SLC26A4 and thus decelerate the development of deafness.
- Hearing loss /
- Deafness /
- miR-26a-5p /
- SLC26A4

HTML全文

图 1 听力损伤和耳聋小鼠模型的鉴定

ABR检测听力损伤和耳聋小鼠的听力阈值（n = 6），^*P < 0.05，^**P < 0.01，^***P < 0.001。

Figure 1. Identification of mouse model of hearing loss and deafness

下载: 全尺寸图片幻灯片

图 2 miR-26a-5p和SLC26A4在听力损伤和耳聋小鼠耳蜗中的表达

A：miR-26a-5p在听力损伤和耳聋小鼠耳蜗中的相对表达量（n = 6）；B：SLC26A4在听力损伤和耳聋小鼠耳蜗中的相对表达量（n = 6）。^*P < 0.05，^**P < 0.01。

Figure 2. Expression of miR-26a-5p and SLC26A4 in the cochlea of mice with hearing impairment and deafness

下载: 全尺寸图片幻灯片

图 3 miR-26a-5p可靶向SLC26A4调控其表达

A：miR-26a-5p和SLC26A4的结合位点预测；B：荧光素酶报告基因实验；C：miR-26a-5p的antagomir和agomir注射小鼠后miR-26a-5p的相对表达量（n = 6）；D：miR-26a-5p的antagomir和agomir注射小鼠后SLC26A4的相对表达量（n = 6）。^*P < 0.05，^**P < 0.01，^***P < 0.001。

Figure 3. miR-26a-5p targets SLC26A4 and regulates its expression

下载: 全尺寸图片幻灯片

图 4 miR-26a-5p调控SLC26A4达到改善小鼠听力减退

A： SLC26A4干扰和过表达载体注射听力损失小鼠后的听力阈值（n = 6）；B：miR-26a-5p干扰和过表达载体注射听力损失小鼠后的听力阈值（n = 6）。^*P < 0.05，^**P < 0.01。

Figure 4. miR-26a-5p regulates SLC26A4 to improve hearing loss in mice

下载: 全尺寸图片幻灯片

表 1 引物序列

Table 1. Primer sequence

基因名称	上游引物序列（5′- 3′）	下游引物序列（5′- 3′）
β-actin	GTGGGGCGCCCCAGGCACCA	CTCCTTAATGTCACGCACGATTT
SLC26A4	GCATCCTCTCCATTATCTACA	TCCTTAACAGCCATACAGAC
miR-26a-5p	通用引物	AGCCAGCGTTCAAGTAATCCAG
U6	通用引物	ATGGACTATCATATGCTTACCGTA

下载: 导出CSV

参考文献(16)

[1]	杨慧波,虞闰六,缪勤,等. 新生儿耳聋基因检测结果分析[J]. 昆明医科大学学报,2020,41(7):144-148. doi: 10.3969/j.issn.1003-4706.2020.07.029
[2]	Harris C,Hemer S,Chur-Hansen A. Emotion as motivator: Parents,professionals and diagnosing childhood deafness[J]. Medical Anthropology,2021,40(3):254-266. doi: 10.1080/01459740.2020.1796659
[3]	Aifa-Hmani M,Ayadi H. Deafness genes and the mechanism of hearing[J]. Archives de L'institut Pasteur de Tunis,2000,77(1-4):17-21.
[4]	Knipper M,Van Dijk P,Schulze H,et al. The neural bases of tinnitus:lessons from deafness and cochlear implants[J]. Journal of Neuroscience,2020,40(38):7190-7202. doi: 10.1523/JNEUROSCI.1314-19.2020
[5]	涂云贵,杜琼,余映辉,等. 云南省安宁地区184例耳聋基因筛查的结果[J]. 昆明医科大学学报,2019,40(12):44-48. doi: 10.3969/j.issn.1003-4706.2019.12.010
[6]	郝帅, 于飞, 杨博, 等. D-半乳糖对仔鼠脂质过氧化、听力和耳蜗毛细胞的损伤及其机制. 吉林大学学报（医学版）, 2012, 38（3）: 419-422.
[7]	Romano D R,Hashino E,Nelson R F. Deafness-in-a-dish: modeling hereditary deafness with inner ear organoids[J]. Human Genetics,2022,141(3-4):347-362. doi: 10.1007/s00439-021-02325-9
[8]	Hardelin J P,Denoyelle F,Levilliers J,et al. Hereditary deafness: Molecular genetics[J]. Medecine Sciences:M/S,2004,20(3):311-316. doi: 10.1051/medsci/2004203311
[9]	Ho P T B,Clark I M,Le L T T. MicroRNA-based diagnosis and therapy[J]. International Journal of Molecular Sciences,2022,23(13):7167. doi: 10.3390/ijms23137167
[10]	Krol J,Loedige I,Filipowicz W. The widespread regulation of microRNA biogenesis,function and decay[J]. Nature reviews genetics,2010,11(9):597-610. doi: 10.1038/nrg2843
[11]	Pritchard C C,Cheng H H,Tewari M. MicroRNA profiling: approaches and considerations[J]. Nature Reviews Genetics,2012,13(5):358-369. doi: 10.1038/nrg3198
[12]	Friedman L M,Avraham K B. MicroRNAs and epigenetic regulation in the mammalian inner ear: implications for deafness[J]. Mammalian Genome,2009,20(9-10):581-603. doi: 10.1007/s00335-009-9230-5
[13]	Mahmoodian-Sani M R,Mehri-Ghahfarrokhi A. The potential of miR-183 family expression in inner ear for regeneration,treatment,diagnosis and prognosis of hearing loss[J]. Journal of Otology,2017,12(2):55-61. doi: 10.1016/j.joto.2017.03.003
[14]	Tawalbeh M,Aburizeg D,Abu Alragheb B O,et al. SLC26A4 phenotypic variability influences intra- and inter-familial diagnosis and management[J]. Genes,2022,13(12):2192. doi: 10.3390/genes13122192
[15]	Hilgert N,Smith R J H,Van Camp G. Forty-six genes causing nonsyndromic hearing impairment: which ones should be analyzed in DNA diagnostics?[J]. Mutation Research/Reviews in Mutation Research,2009,681(2-3):189-196. doi: 10.1016/j.mrrev.2008.08.002
[16]	Park H J,Shaukat S,Liu X Z,et al. Origins and frequencies of SLC26A4 (PDS) mutations in east and south Asians:global implications for the epidemiology of deafness[J]. Journal of Medical Genetics,2003,40(4):242-248. doi: 10.1136/jmg.40.4.242

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