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Full Length Article | Open Access

Beclin-1/LC3-Ⅱ dependent macroautophagy was uninfluenced in ischemia-challenged vascular endothelial cells

Yaping Maa,1, Chaofan Lib,1, Yan Hec,d,1, Tiwei Fue, Li Songa, Qingsong Yed,f, Fugui Zhangb,d()
Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing 401147, PR China
Department of Oral and Maxillofacial Surgery, Stomatological Hospital of Chongqing Medical University, Chongqing 401147, PR China
Tianyou Hospital, Wuhan University of Science & Technology, Wuhan, Hubei 430064, PR China
Department of Oral and Maxillofacial Surgery, Massachusetts General Hospital, Boston, MA 02114, USA
Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, PR China
Center for Regenerative Medicine, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, PR China

Peer review under responsibility of Chongqing Medical University.

1 These authors made equal contributions to this study.

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Abstract

Autophagy has been extensively studied and occurs in many biological settings. However, a question remains as to whether ischemia enhances Beclin-1/LC3-Ⅱ-dependent macroautophagy in vascular endothelial cells, as has been previously thought. Furthermore, the effect of the level of autophagy on cell or skin flap survival still requires elucidation. We created a lethal ischemia model in human umbilical vascular endothelial cells (HUVECs), performed quantitative proteomics and bioinformatics analyses, and verified the autophagic status and effect both in vitro and in vivo. The significantly upregulated proteins encoded by autophagy-related genes (ATGs) included ATG2A, ATG3, ATG4B, ATG5, ATG7, ATG9A, ATG12, ATG16, and ATG101. The significantly enhanced lysosomal proteins were cathepsin B, cathepsin D, lysosome-associated membrane protein 1 (LAMP1), and LAMP2. However, the differentially expressed proteins excluded Beclin-1, microtubule-associated protein light chain 3 (LC3)-Ⅰ, and LC3-Ⅱ. Western blot analyses verified that the protein expression levels of Beclin-1, LC3-Ⅰ, and LC3-Ⅱ were neither upregulated nor downregulated in ischemia-challenged HUVECs. The autophagic status was not enhanced by rapamycin in ischemic HUVECs but appeared to be inhibited by chloroquine. Our in vivo study on rats showed that a downregulation in autophagic status jeopardized skin flap survival. In conclusion, Ischemia neither enhanced nor inhibited Beclin-1/LC3-Ⅱ-dependent canonical macroautophagy both in vitro and in vivo, in contradiction to previous studies. An appropriate autophagic homeostasis can minimize cell or skin flap damage.

Keywords

AutophagyBioinformatics analysisHuman umbilical vein endothelial cellQuantitative proteomics studySkin flap

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Genes & Diseases
Volume 9 Issue 2,
March 2022
Pages 549-561
DOI: 10.1016/j.gendis.2021.02.010
Cite this article:
Ma Y, Li C, He Y, et al. Beclin-1/LC3-Ⅱ dependent macroautophagy was uninfluenced in ischemia-challenged vascular endothelial cells. Genes & Diseases, 2022, 9(2): 549-561. https://doi.org/10.1016/j.gendis.2021.02.010
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