甲苯对大鼠耳蜗外毛细胞蛋白表达谱的影响

职业与健康 ›› 2024, Vol. 40 ›› Issue (23): 3188-3193.

甲苯对大鼠耳蜗外毛细胞蛋白表达谱的影响

张岚^1,2, 廖慧², 黎宗男³, 袁建辉^{1, 2, 3}

1.南方医科大学公共卫生学院,广东广州 510515;
2.深圳市南山区疾病预防控制中心职业卫生科,广东深圳 518054;
3.广东医科大学公共卫生学院,广东东莞 523000

收稿日期:2024-04-11 修回日期:2024-09-23 出版日期:2024-12-01 发布日期:2026-04-09
通信作者: 袁建辉,主任技师,E-mail：jianhui_yuan@126.com
作者简介:张岚,女,在读硕士研究生,研究方向为职业卫生与中毒控制。
基金资助:
深圳市南山区科技计划项目（NS2021034）

Effects of toluene on the protein expression profiles of rat cochlear outer hair cells

ZHANG Lan^1,2, LIAO Hui², LI Zongnan³, YUAN Jianhui^{1, 2, 3}

1. School of Public Health,Southern Medical University,Guangzhou,Guangdong 510515,China;
2. Department of Occupational Health,Shenzhen Nanshan District Center for Disease Control and Prevention,Shenzhen,Guangdong 518054,China;
3. School of Public Health,Guangdong Medical University,Dongguan,Guangdong 523000,China

Received:2024-04-11 Revised:2024-09-23 Online:2024-12-01 Published:2026-04-09
Contact: YUAN Jianhui,Chief technician,E-mail：jianhui_yuan@126.com

摘要/Abstract

摘要： 目的筛选甲苯暴露后大鼠耳蜗外毛细胞中的差异表达蛋白,分析差异表达蛋白的功能及相互作用。方法用含不同剂量,浓度分别为0、5、10、20、40、50、60、80和100 mmol/L甲苯的1640基础培养基处理大鼠耳蜗外毛细胞,CCK-8检测细胞活性,通过QE质谱仪检测染毒组与对照组大鼠耳蜗外毛细胞的蛋白表达谱变化,Perseus软件分析差异表达蛋白,基因本体（gene ontology,GO）分析和京都基因与基因组百科全书（kyoto encyclopedia of genes and genomes,KEGG）富集分析差异表达蛋白的功能,蛋白免疫印迹法（western blot,WB）验证hub蛋白的表达。结果共筛选到233个差异表达蛋白,其中上调蛋白157个,下调蛋白76个,GO分析显示,差异蛋白的功能主要与糖酵解、糖异生和糖蛋白结合过程有关;KEGG富集分析主要涉及糖酵解/糖异生、细胞外泌体途径的失调。通过对失调通路糖酵解/糖异生中的蛋白进行蛋白质相互作用（protein-protein interaction,PPI）分析筛选得到磷酸丙糖异构酶1（triosephosphate isomerase 1,TPI1）、磷酸甘油酸激酶1（phosphoglycerate kinase 1,PGK1）和果糖二磷酸醛缩酶A（fructose-bisphosphate aldolase A,ALDOA）3个关键蛋白。经甲苯染毒处理后大鼠耳蜗外毛细胞中TPI1、PGK1和ALDOA蛋白表达量均降低。结论甲苯暴露可能通过抑制TPI1、PGK1和ALDOA蛋白的表达而引起糖代谢途径失调,导致大鼠耳蜗细胞损伤。

关键词: 甲苯, 大鼠耳蜗外毛细胞, 差异表达蛋白

Abstract: Objective To filter the differentially expressed proteins（DEPs） in outer hair cells（OHCs） of rat cochlear after toluene exposure,and analyze the functions and interactions of DEPs. Methods Rat cochlear OHCs were cultured in the 1640 basal medium containing toluene at different doses（concentrations：0,5,10,20,40,50,60,80,and 100 mmol/L）. Cell activity was detected by CCK-8,and the changes of protein expression profile of rat cochlear OHCs in the exposed group and the control group were detected by Q Extractive mass spectrometry. DEPs were analyzed by Perseus software and their biological functions were analyzed by Gene Ontology（GO） and Kyoto Encyclopedia of Genes and Genomes（KEGG） enrichment. The expression of hub proteins was detected by Western Blot（WB）. Results A total of 233 DEPs were screened,including 157 up-regulated proteins and 76 down-regulated proteins. GO analysis results showed that the functions of DEPs were mainly involved in glycolytic,gluconeogenesis and glycoprotein binding processes. KEGG enrichment analysis showed that DEPs were mainly related to dysregulation of glycolysis/gluconeogenesis and extracellular exosome pathways. Three key proteins,namely triosephosphate isomerase 1（TPI1）,phosphoglycerate kinase 1（PGK1） and fructose-bisphosphate Aldolase A（ALDOA）,were identified through protein-protein interaction（PPI） analysis and screening of proteins involved in the dysregulated pathway of glycolysis/gluconeogenesis. WB results showed that TPI1,PGK1 and ALDOA protein expression levels in rat cochlear OHCs were decreased after toluene exposure. Conclusion Exposure to toluene may cause dysregulation of glucose metabolism pathways and lead to cochlear cell damage in rats by inhibiting the expression of TPI1,PGK1,and ALDOA proteins.

Key words: Toluene, Outer hair cells of rat cochlear, Differentially expressed proteins

中图分类号:

R-332

张岚, 廖慧, 黎宗男, 袁建辉. 甲苯对大鼠耳蜗外毛细胞蛋白表达谱的影响. [J]职业与健康, 2024, 40(23): 3188-3193.

ZHANG Lan, LIAO Hui, LI Zongnan, YUAN Jianhui. Effects of toluene on the protein expression profiles of rat cochlear outer hair cells. [J]OCCUPATION AND HEALTH, 2024, 40(23): 3188-3193.

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