Benzo(a)pyrene blocking up the autophagic flux in human umbilical vein endothelial cell by down-regulating the expressions of syntaxin 17 and lysosomal associated membrane protein 2
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1.School of Public Health of University of South China & Hengyang Key Laboratory of New Technology for Inspection and Quarantine of Health Hazard Factors, Hengyang, Hunan 421001, China;2.Institute of Preventive Medicine, School of Public Health, Guilin Medical University, Guilin, Guangxi 541199, China;3.Guangzhou Institute of Geochemistry, Chinese Academy of Sciences & State Key Laboratory of Organic Geochemistry & Guangdong Key Laboratory of Environmental Resources Utilization and Protection, Guangzhou, Guangdong 510640, China)

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R54

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    Abstract:

    Aim To study the mechanism of benzo(a)pyrene (BaP) affecting autophagy in human umbilical vein endothelial cell (HUVEC). Methods HUVECs were treated with BaP (2.5,5, 10 μmol/L) for 24 h. The degradation of autophagosome and its contents, the expressions of target proteins microtubule-associated protein 1 light chain 3 (LC3), sequestosome 1 (p62), Beclin-1, autophagy-related protein 5 (Atg5), Atg7, Atg12, cathepsin B (CTSB), cathepsin D (CTSD), syntaxin 17 (STX17), lysosomal associated membrane protein 2 (LAMP2), the number and function of lysosomes, and the phosphorylation levels of related upstream key regulatory proteins serine-threonine protein kinase (Akt), extracellular regulated protein kinase (ERK) and transcription factor EB (TFEB) were detected respectively by indirect immunofluorescence, Western blot, acridine orange staining and monodansyl cadaverine staining.Results In HUVEC of BaP exposure group, the test results showed that:(1)The level of LC3 puncta and the ratio of LC3Ⅱ/LC3Ⅰ increased, the expressions of key autophagy initiation proteins (Beclin-1, Atg5, Atg7 and Atg12) increased, and the phosphorylation of Akt protein decreased significantly; (2)The levels of p62 puncta and p62 protein increased significantly; (3)The number of lysosomes increased, accompanied by the increased expressions of lysosomal characteristic hydrolases (CTSB, CTSD), and the phosphorylation levels of ERK and TFEB increased accordingly; (4)The key proteins STX17 and LAMP2 regulating the fusion of autophagy and lysosome decreased. Conclusion BaP inhibits the normal autophagic flux of HUVEC by reducing the expression levels of STX17 and LAMP2.

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ZHAO Dongting, PENG Wenyi, XUE Panpan, JIANG Xiaojun, CHEN Shuting, GAO Huiqian, FENG Shaolong. Benzo(a)pyrene blocking up the autophagic flux in human umbilical vein endothelial cell by down-regulating the expressions of syntaxin 17 and lysosomal associated membrane protein 2[J]. Editorial Office of Chinese Journal of Arteriosclerosis,2021,29(11):934-940.

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  • Received:December 25,2020
  • Revised:April 08,2021
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  • Online: November 18,2021
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