Cx30基因的慢病毒质粒构建及其对高糖环境耳蜗毛细胞活力影响
Journal: Basic Medical Theory Research DOI: 10.32629/bmtr.v8i2.18917
Abstract
目的：构建过表达Cx30基因的慢病毒质粒，研究Cx30对高糖诱导耳蜗毛细胞凋亡、增殖影响。方法：本研究通过慢病毒载体构建了携带Flag标签过表达CX30的复合质粒，利用瞬时转染技术过表达CX30、通过CCK-8实验检测细胞活力，流式细胞术检测细胞线粒体膜电位与细胞凋亡。结果：成功构建CX30真核表达质粒。在HEI-OC1细胞中建立NC组和过表达CX30组，在25mmol/L条件下，NC和CX30组细胞数差异显著（P<0.01），CCK-8实验显示，在25mmol/L浓度时，CX30组OD490nm高于NC组（P<0.05）。流式细胞检测细胞凋亡：在25mmol/L浓度时，NC组晚期凋亡率3.4%；CX30组晚期凋亡率4.72%。激光共聚焦显微镜成像显示CX30与PLVAP空间共定位于细胞质及细胞膜。结论：过表达CX30可能通过调控PLVAP抑制凋亡，减轻高糖诱导耳蜗毛细胞损伤。
Keywords
CX30；慢病毒包装；细胞凋亡；增殖；HEI-OC1
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[3] ALADAG I, EYIBILEN A, GüVEN M, et al. Role of oxidative stress in hearing impairment in patients with type two diabetes mellitus [J]. J Laryngol Otol, 2009, 123(9): 957-63.
[4] YADAV M K, YADAV K S. Etiology of Noise-Induced Hearing Loss (NIHL) and its Symptomatic Correlation with Audiometry Observations in Type II Diabetes [J]. Indian J Otolaryngol Head Neck Surg, 2018, 70(1): 137-44.
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[9] 张初琴, 陈波蓓, 伊松, 等. 非综合征型耳聋的不同听力学表型与常见致聋基因的相关性[J]. 温州医科大学学报, 2020, 50(11): 873-7.
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[12] XU J, NICHOLSON B J. Divergence between Hemichannel and Gap Junction Permeabilities of Connexin 30 and 26 [J]. Life (Basel), 2023, 13(2).
[13] LEVIN M, MERCOLA M. Expression of connexin 30 in Xenopus embryos and its involvement in hatching gland function [J]. Dev Dyn, 2000, 219(1): 96-101.
[14] MCCULLOCH F, CHAMBREY R, ELADARI D, et al. Localization of connexin 30 in the luminal membrane of cells in the distal nephron [J]. Am J Physiol Renal Physiol, 2005, 289(6): F1304-12.
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[19] PACIELLO F, ZORZI V, RASPA M, et al. Connexin 30 deletion exacerbates cochlear senescence and age-related hearing loss [J]. Front Cell Dev Biol, 2022, 10: 950837.
[20] CHEN J, CHEN P, HE B, et al. Connexin30-Deficiency Causes Mild Hearing Loss With the Reduction of Endocochlear Potential and ATP Release [J]. Front Cell Neurosci, 2021, 15: 819194.
[21] CHEN B, XU H, MI Y, et al. Mechanisms of hearing loss and cell death in the cochlea of connexin mutant mice [J]. Am J Physiol Cell Physiol, 2020, 319(3): C569-c78.
[22] LERMAN-GARBER I, CUEVAS-RAMOS D, VALDéS S, et al. Sensorineural hearing loss--a common finding in early-onset type 2 diabetes mellitus [J]. Endocr Pract, 2012, 18(4): 549-57.
[23] 李晋莹. 2型糖尿病患者的血糖在目标范围内的时间与听力损害的相关性研究[D]. 暨南大学, 2023.
[24] DENG Y, CHEN S, HU J. Diabetes mellitus and hearing loss [J]. Mol Med, 2023, 29(1): 141.
[25] GIRAUDET F, MULLIEZ A, DE RESENDE L M, et al. Impaired auditory neural performance, another dimension of hearing loss in type-2 diabetes mellitus [J]. Diabetes Metab, 2022, 48(6): 101360.
[26] LIU L, XU Q, ZHU M, et al. An Umbrella Review of Diabetes Mellitus and Hearing Impairment [J]. J Evid Based Med, 2025, 18(2): e70030.
[27] XIA A, KATORI Y, OSHIMA T, et al. Expression of connexin 30 in the developing mouse cochlea [J]. Brain Res, 2001, 898(2): 364-7.
[28] SCHüTZ M, SCIMEMI P, MAJUMDER P, et al. The human deafness-associated connexin 30 T5M mutation causes mild hearing loss and reduces biochemical coupling among cochlear non-sensory cells in knock-in mice [J]. Hum Mol Genet, 2010, 19(24): 4759-73.
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