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  • 毛睿,徐晶莹,吕立夏,等.代谢组学方法分析香烟烟雾暴露对大鼠视网膜的影响[J].同济大学学报(医学版),2021,42(1):19-27,34.    [点击复制]
  • MAO Rui,XU Jing-ying,L Li-xia,et al.Effects of exposure to cigarette smoke on retina in rats: a metabolomics study[J].同济大学学报(医学版),2021,42(1):19-27,34.   [点击复制]
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代谢组学方法分析香烟烟雾暴露对大鼠视网膜的影响
毛睿,徐晶莹,吕立夏,崔婷婷,刘彩莹,徐国彤
0
(同济大学医学院,上海200092;同济大学医学院,上海200093;同济大学医学院,上海200094;同济大学医学院,上海200095;同济大学医学院,上海200096;同济大学医学院,上海200097)
摘要:
目的通过构建香烟暴露大鼠模型,并利用代谢组学的方法分析香烟暴露对大鼠视网膜的影响,寻找香烟烟雾中引起视网膜功能下降的潜在毒物。方法使用代谢组学方法分析大鼠经香烟暴露后血浆、房水和视网膜中的差异代谢物。使用LC-MS明确香烟暴露大鼠视网膜中萘代谢产物的存在。使用细胞衰老β-半乳糖苷酶染色(senescence-associated β-galactosidase, SA-β-gal)法检测了香烟暴露大鼠视网膜切片细胞衰老情况。使用闪光视网膜电图(flicker electroretinograms, F-ERG)实验评价大鼠视网膜功能。采用CCK-8法检测1,2-二羟基萘酚对视网膜细胞的毒性作用。结果模型大鼠经香烟暴露后,在其视网膜内发现了萘代谢物1,2-二羟基萘酚与1,2-萘醌。随着暴露的进行,模型大鼠视网膜组织内丙二醛水平升高(P<0.05),总抗氧化能力下降(P<0.05)。香烟暴露大鼠视网膜在4个月时F-ERG检查b波值下降(P<0.05)。约10μmol/L毒物1,2-二羟基萘在体外作用48h可以导致半数aRPE19细胞与r28细胞发生死亡。结论香烟暴露会导致SD大鼠视网膜中氧化应激水平升高,加速视网膜细胞发生衰老,进而引起视网膜功能下降。其中香烟烟雾中的萘是引起大鼠视网膜功能下降的关键物质之一,其体内代谢物1,2-二羟基萘会对视网膜细胞产生很强的细胞毒性作用。
关键词:  香烟暴露  视网膜  萘代谢物  代谢组学  年龄相关性黄斑变性
DOI:10.12289/j.issn.1008-0392.20458
投稿时间:2020-10-27
基金项目:上海市(新增)科学技术委员会自然基金(2017YFA0104100);国家重大科学研究计划(2016YFA0101302)
Effects of exposure to cigarette smoke on retina in rats: a metabolomics study
MAO Rui,XU Jing-ying,L Li-xia,CUI Ting-ting,LIU Cai-ying,LIU Cai-ying
(Tongji University School of Medicine, Shanghai 200092, China;Tongji University School of Medicine, Shanghai 200093, China;Tongji University School of Medicine, Shanghai 200094, China;Tongji University School of Medicine, Shanghai 200095, China;Tongji University School of Medicine, Shanghai 200096, China;Tongji University School of Medicine, Shanghai 200097, China)
Abstract:
ObjectiveTo investigate the effects of cigarette smoke exposure on the retina of rats with metabolomics study. MethodsCigarette smoke exposure model was established in SD rats. Metabolomics was used to analyze the differential metabolites in plasma, aqueous humor and retina of rats after cigarette smoke exposure. LC-MS was used to clarify the presence of naphthalene metabolites in the retina of rats exposed to cigarette smoke. The SA-β-gal method was used to detect the senescence of retinal slices in rats exposed to cigarettes. Flash electroretinogram (F-ERG) experiment was performed to evaluate rat retinal function. The CCK-8 method was used to detect the toxicity of 1,2-dihydroxynaphthol on retinal cells. ResultsAfter the model rats were exposed to cigarette smoke, the naphthalene metabolites 1,2-dihydroxynaphthol and 1,2-naphthoquinone were detected in the retinas. Cigarette exposure accelerates the senescence of rat retinal cells. As the exposure progressed, the level of malondialdehyde in the retina of the model rats increased (P<0.05), and the total antioxidant capacity decreased (P<0.05). The b-wave value of F-ERG in the retina of rats decreased 4 months after cigarette smoke exposure (P<0.05). About 10μmol/L poison 1,2-dihydroxynaphthalene induced half of aRPE19 cells and r28 cells to die in vitro after exposure for 48h. ConclusionCigarette smoke exposure will increase the level of oxidative stress in the retina of SD rats, accelerate the senescence of retinal cells, and cause the decline of retinal function. Among them, naphthalene in cigarette smoke is one of the key substances that cause the decline of retinal function in rats, and its metabolite 1,2-dihydroxynaphthalene has a strong cytotoxic effect on retinal cells.
Key words:  cigarette exposure  retina  naphthalene metabolites  metabolomics  age-related macular degeneration

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