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Loss of HMBOX1 promotes LPS-induced apoptosis and inhibits LPS-induced autophagy of vascular endothelial cells in mouse

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Abstract

Our previous study revealed that Homeobox containing 1 (HMBOX1), essential for the survival of vascular endothelial cells (VECs), was involved in the progression of atherosclerosis. Knockdown of HMBOX1 promoted apoptosis and inhibited autophagy through regulating intracellular free zinc level in cultured VECs. In current study, in order to investigate the roles of HMBOX1 in vivo and in endothelium, we generated a knockout (KO) mouse for HMBOX1 by using transcription activator-like effector nucleases (TALENs) technology. Herein, we reported that the protein level of HMBOX1 was gradually increased during mouse development. The HMBOX1 KO mouse was viable and fertile. There existed no differences in apoptosis and autophagy of aortic endothelial cells between wild type and KO mouse. Whereas, loss of HMBOX1 promoted apoptosis and inhibited autophagy of aortic endothelial cells under lipopolysaccharide (LPS) stimulation in mouse. We also demonstrated that HMBOX1 deletion had no influence on the secretion of inflammatory cytokines TNF-α and IL-6. Moreover, overexpression or knockdown of HMBOX1 failed to regulate multiple pro-apoptotic genes expression in vitro. In conclusion, HMBOX1 participated in the functional maintenance of mouse aortic endothelial cells, the aortic endothelial cells of HMBOX1 KO mouse showed increased apoptosis and decreased autophagy with LPS treatment.

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Abbreviations

ALT:

Alternative lengthening of telomeres

BMSC:

Bone marrow mesenchymal stem cell

ESC:

Embryonic stem cells

HMBOX1:

Homeobox containing 1

HNF:

Hepatocyte nuclear factor

HUVEC:

Human umbilical vascular endothelial cell

LPS:

Lipopolysaccharide

MT2A:

Metallothionein 2A

TALEN:

Transcription activator-like effector nuclease

VEC:

Vascular endothelial cell

WT:

Wild type

KO:

Knockout

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Acknowledgements

This work was financially supported by National Natural Science Foundation of China (Grant Nos. 31870831, 91539105, 81321061, 81902656), and the Shandong Provincial Natural Science Foundation (Grant No. ZR2019BH059).

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Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, Qingdao, 266237, People’s Republic of China

    HanLin Ma, Le Su, XiaoYing He & JunYing Miao

  2. Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, Jinan, 250012, People’s Republic of China

    HanLin Ma

  3. The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Shandong University Qilu Hospital, Jinan, 250012, People’s Republic of China

    JunYing Miao

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  1. HanLin Ma
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  2. Le Su
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Contributions

MH conceived of the presented idea, designed and performed the experiments, analyzed the data, carried out data interpretation, designed the figures, wrote the paper. SL performed the experiments, analyzed the data, carried out data interpretation. HX performed the experiments. MJ conceived and designed the experiments, supervised the work. All authors read and approved the final version of this manuscript.

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Correspondence to JunYing Miao.

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All experimental procedures and animal care were performed in accordance with the ARRIVE guidelines [38] and approved by the ethics committee of Shandong University.

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Ma, H., Su, L., He, X. et al. Loss of HMBOX1 promotes LPS-induced apoptosis and inhibits LPS-induced autophagy of vascular endothelial cells in mouse. Apoptosis 24, 946–957 (2019). https://doi.org/10.1007/s10495-019-01572-6

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