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引用:姚国敏,古丽努尔·买买提,李蓉,邓颖,焦聪.脂联素对视网膜血管内皮细胞缺氧损伤的保护作用及机制研究.国际眼科杂志 2020;20(8):1320-1324,doi:10.3980/j.issn.1672-5123.2020.8.05
脂联素对视网膜血管内皮细胞缺氧损伤的保护作用及机制研究
Study on the protective effect and mechanism of adiponectin on hypoxic injury of retinal vascular endothelial cells
投稿时间:2020-03-04  修订日期:2020-06-29
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DOI:10.3980/j.issn.1672-5123.2020.8.05
关键词:  脂联素  RF/6A细胞  缺氧  氧化应激  细胞活力  凋亡
Key Words:  adiponectin  RF/6A cell  hypoxia  oxidative stress  cell viability  apoptosis
基金项目:陕西省卫生健康科研基金项目(No.2018D074); 陕西省科技厅自然科学基金配套基金(No.XYFYPT-2020-07)
Fund Project:Health Research Foundation of Shaanxi Province(No.2018D074); Matching Foundation of Shaanxi Provincial Science and Technology Department(No.XYFYPT-2020-07)
              
作者单位
姚国敏 中国陕西省西安市,西安医学院第一附属医院眼科
古丽努尔·买买提 中国新疆维吾尔自治区乌鲁木齐市,新疆维吾尔自治区人民医院眼科
李蓉 中国陕西省西安市,西安医学院第一附属医院眼科
邓颖 中国陕西省西安市,西安医学院第一附属医院眼科
焦聪 中国陕西省西安市,西安医学院第一附属医院眼科
              
AuthorInstitution
Guo-Min Yao Department of Ophthalmology, the First Affiliated Hospital of Xi'an Medical University, Xi'an , Shaanxi Province, China
Gulinuer Maimaiti Department of Ophthalmology, People's Hospital of Xinjiang Uygur Autonomous Region, Urumchi , Xinjiang Uygur Autonomous Region, China
Rong Li Department of Ophthalmology, the First Affiliated Hospital of Xi'an Medical University, Xi'an , Shaanxi Province, China
Ying Deng Department of Ophthalmology, the First Affiliated Hospital of Xi'an Medical University, Xi'an , Shaanxi Province, China
Cong Jiao Department of Ophthalmology, the First Affiliated Hospital of Xi'an Medical University, Xi'an , Shaanxi Province, China
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目的:研究脂联素对缺氧损伤的恒河猴脉络膜/视网膜内皮(RF/6A)细胞的保护作用及相关机制。

     方法:将体外培养的RF/6A细胞随机分为对照组、缺氧损伤(CoCl2刺激诱导24h)组和缺氧损伤+脂联素(5、50、100μmol/L脂联素预处理6h)组。采用MTT法检测细胞活力,选择合适的脂联素内皮细胞保护作用浓度,并采用Western blot检测凋亡相关因子Bax、Bcl-2的表达情况,同时测定细胞内活力氧簇(ROS)的表达水平。

     结果:与对照组相比,缺氧损伤组和各浓度脂联素预处理的RF/6A细胞活力均下降(均P<0.01); 与缺氧损伤组相比,各浓度脂联素预处理后细胞活力均显著增加(均P<0.05),其中50μmol/L脂联素是较为合适的保护作用浓度。与对照组相比,缺氧损伤组细胞活力降低,Bax蛋白表达量升高,Bcl-2蛋白表达量降低,ROS生成增加(P<0.01); 与缺氧损伤组相比,脂联素预处理后细胞活力增加,Bax蛋白表达量降低,Bcl-2蛋白表达量增加,ROS生成减少(P<0.01)。

     结论:脂联素可明显减轻缺氧造成的视网膜血管内皮细胞损伤和凋亡,其机制可能与脂联素减轻氧化应激有关。

Abstract:
      AIM:To investigate the protective effect of adiponectin on hypoxia-damaged rhesus monkey choroid /retinal vascular endothelial cells(RF/6A)and related mechanisms.

     METHODS:In vitro cultured RF/6A cells were randomly divided into the control group, hypoxic injury(induced by CoCl2 stimulation)group and hypoxic injury + adiponectin(5μmol/L, 50μmol/L and 100μmol/L)group. Cell viability was assessed using the MTT assay and optimal concentration of adiponectin was selected. Western blot was used to detect the expression of Bax and Bcl-2 in RF/6A cells. Reactive oxygen species(ROS)detection kit was used to detect the content of ROS in RF/6A cells.

     RESULTS: Compared with the control group, the cell viability of RF/6A cells in the hypoxic injury group and each adiponectin pretreatment group decreased(all P<0.01). Compared with the hypoxic injury group, the cell viability of RF/6A cells in each adiponectin pretreatment group was significantly increased(all P<0.05), and adiponectin of 50μmol/L was the appropriate protective concentration. Compared with the control group, the viability of RF/6A cells decreased, the protein expression level of Bax increased, the protein expression level of Bcl-2 decreased, and the content of ROS increased in the hypoxic injury group(all P<0.01). Compared with the hypoxic injury group, the viability RF/6A cells increased, the expression level of Bax decreased, the expression level of Bcl-2 increased, and the content of ROS decreased in the adiponectin pretreatment group(all P<0.01).

     CONCLUSION: Our findings suggest that adiponectin can significantly alleviate retinal vascular endothelial cell damage and apoptosis caused by hypoxia, and the mechanism may be related to the inhibition of oxidative stress by adiponectin.

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