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

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.

Therapeutic angiogenesis for patients with no-option critical limb ischemia by adipose-derived regenerative cells: TACT-ADRC multicenter trial

“…The Graph-Pad Prism software version 8.0 (GraphPad Software Inc) was used. Values of p < 0.05 were considered statistically significant [20].…”

Hydrogen Sulfide Attenuates Lymphedema Via the Induction of Lymphangiogenesis Through a PI3K/Akt‐Dependent Mechanism

“…After overnight starvation, the cells were reseeded onto a 6‐well plate coated with Matrigel (Becton Dickinson, Bedford, MA) at 2×10 4 cells per well in EBM‐2 with 0.5% FBS medium. 22 The cells were incubated at 37 °C for 6 hours with DATS or vehicle, and morphological changes of LECs were microscopically analyzed in 3 different fields per well. Tube formation was assessed by the average of total tube lengths formed in the 3 fields per sample using Image J software (version 1.51).…”

New insight into ischemic stroke: Circadian rhythm in post-stroke angiogenesis

“…Following research explored the impact of Per2 in EPCs (endothelial progenitor cells); Per2 −/− EPCs displayed impaired proliferation, migration, tube formation, and adhesion ( Sun et al, 2014 ). Jet lag plays an antiangiogenic role and inhibits blood reperfusion after ischemic injury; further loss-of-function studies explore the relation between Cry and HUVEC (human umbilical vein endothelial cell) phenotypes, which confirmed that knockdown of Cry1 and Cry2 in HUVECs inhibited proliferation, migration, and tubular morphological features ( Tsuzuki et al, 2021 ).…”

“…The human VEGF promoter activity follows a 24-h circadian pattern under the 12 h light–dark (LD) cycle, and the core circadian gene Bmal1 plays a transcriptional role by targeting the region of the VEGF promoter ( Jensen et al, 2012 ). In addition, the fluctuation of the serum VEGF level displayed a rhythmic pattern peaking during noon, while ECD induced by jet lag impaired this fluctuation by decreasing the VEGF serum level ( Tsuzuki et al, 2021 ). Moreover, Per2 and Cry1 act as critical rhythmic regulators of the hypoxia-induced expression of the VEGF; these observations indicate the existence of a negative feedback loop that rhythmically suppresses the transcriptional upregulation of VEGF under hypoxic conditions, resulting in the circadian fluctuation of VEGF expression and angiogenesis ( Koyanagi et al, 2003 ; Figure 4 ).…”