Adverse Effect of Circadian Rhythm Disorder on Reparative Angiogenesis in Hind Limb Ischemia

• Type of Material: Article
• Authors: Kazuhito Tsuzuki, Nagoya University; Yuuki Shimizu, Nagoya University; Junya Suzuki, Nagoya University; Zhongyue Pu, Nagoya University; Shukuro Yamaguchi, Nagoya University; Yusuke Fujikawa, Nagoya University; Katsuhiro Kato, Nagoya University; Koji Ohashi, Nagoya University; Mikito Takefuji, Nagoya University; Yasuko K Bando, Nagoya University; Noriyuki Ouchi, Nagoya University; John Calvert, Emory University; Rei Shibata, Nagoya University; Toyoaki Murohara, Nagoya University
• Language: English
• Date: 2021-08-17
• Publisher: WILEY
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2021 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.
• License: Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1161/JAHA.121.020896
• Title of Journal or Parent Work: JOURNAL OF THE AMERICAN HEART ASSOCIATION
• Volume: 10
• Issue: 16
• Start Page: e020896
• End Page: e020896
• Grant/Funding Information: This work was supported by a grant (No. 17H06745 to Dr Shimizu) from the Ministry of Education, Culture, Sports, Science and Technology of Japan; and a grant from Japan Foundation for Applied Enzymology (No. 14T020 to Dr Shimizu).
• Keywords: angiogenesis; endothelial progenitor cell; CORONARY-HEART-DISEASE; ENDOTHELIAL GROWTH-FACTOR; Cardiac & Cardiovascular Systems; Life Sciences & Biomedicine; cryptochrome; CLOCK; PHYSIOLOGY; hind limb ischemia; RISK; MISALIGNMENT; vascular endothelial growth factor; SHIFT WORK; CELLS; GENE-EXPRESSION; circadian rhythm; Cardiovascular System & Cardiology; Science & Technology; CRYPTOCHROME
• Research Categories: Health Sciences, Medicine and Surgery

Abstract

BACKGROUND: Circadian rhythm disorders, often seen in modern lifestyles, are a major social health concern. The aim of this study was to examine whether circadian rhythm disorders would influence angiogenesis and blood perfusion recovery in a mouse model of hind limb ischemia. METHODS AND RESULTS: A jet-lag model was established in C57BL/6J mice using a light-controlled isolation box. Control mice were kept at a light/dark 12:12 (12-hour light and 12-hour dark) condition. Concentrations of plasma vascular endothelial growth factor and circulating endothelial progenitor cells in control mice formed a circadian rhythm, which was diminished in the jet-lag model (P<0.05). The jet-lag condition deteriorated tissue capillary formation (P<0.001) and tissue blood perfusion recovery (P<0.01) in hind limb ischemia, which was associated with downregulation of vascular endothelial growth factor expression in local ischemic tissue and in the plasma. Although the expression of clock genes (ie, Clock, Bmal1, and Cry) in local tissues was upregulated after ischemic injury, the expression levels of cryptochrome (Cry) 1 and Cry2 were inhibited by the jet-lag condition. Next, Cry1 and Cry2 double-knockout mice were examined for blood perfusion recoveries and a reparative angiogenesis. Cry1 and Cry2 double-knockout mice revealed suppressed capillary density (P<0.001) and suppressed tissue blood perfusion recovery (P<0.05) in the hind limb ischemia model. Moreover, knockdown of CRY1/2 in human umbilical vein endothelial cells was accompanied by increased expression of WEE1 and decreased expression of HOXC5. This was associated with decreased proliferative capacity, migration ability, and tube formation ability of human umbilical vein endothelial cells, respectively, leading to impairment of angiogenesis. CONCLUSIONS: Our data suggest that circadian rhythm disorder deteriorates reparative ischemia-induced angiogenesis and that maintenance of circadian rhythm plays an important role in angiogenesis.