Abstract 006: Liver-specific Bmal1 Deletion In Mice Increases Aortic Stiffness With Age-dependent Alterations In The Vasculature

Abstract

Circadian clock genes are important for vascular homeostasis. Loss of Bmal1 , a clock gene, impairs vascular function and blood pressure rhythm in mice. We previously reported that hepatocyte-specific Bmal1 deletion (HBK) in the liver alters perivascular adipose tissue-mediated vascular function in young adult mice, yet aortic collagen content and wall thickness in 4- to 6- month old HBK mice is similar to control genotype flox mice. To our knowledge, this is some of the first evidence that liver circadian clock disruption distally affects function in another tissue. We hypothesized that Bmal1 deletion in liver leads to vascular disease in older adult mice. Studies were performed in 8- to 11-month old male HBK and flox control mice. Aortic stiffness, measured by pulse wave velocity, was significantly higher in HBK mice compared to flox control mice (Flox: 1.93 ± 0.2 m/s; HBK: 3.3 ± 0.5 m/s; n = 7-8, p = 0.02). Light phase systolic blood pressure (tail-cuff) was similar in both flox control and HBK mice (Flox: 101 ± 1 mm Hg; HBK: 103 ± 2 mm Hg; n = 3-5, p = 0.35). Plasma and aortas were collected at ZT10 for metabolite measurements and histological analysis. Circulating plasminogen activator inhibitor-1 (PAI-1) was not different between genotypes. Picrosirius red (PSR)-stained aortic sections were examined under bright field or polarized light to assess collagen content with Metamorph software analysis. Aortic collagen content was not different between flox control and HBK mice under bright or polarized light (bright light, % area stained positive for PSR, Flox: 28.5 ± 2.4%; HBK: 23.6 ± 3.3%, p = 0.30; polarized, Flox: 16.7 ± 0.8%; HBK: 16.3 ± 1.3%; n = 4-5, p=0.82). TUNEL staining showed increased cellular apoptosis in aortas of HBK mice (Flox: 0.72 ± 0.3%; HBK: 2.73 ± 0.7%; n = 4, p = 0.04). Aortic wall thickness was measured as the difference between the external elastic lamina and the internal elastic lamina with CellSens software. Interestingly, aortic wall thickness was significantly lower in older HBK mice compared to flox control mice (Flox: 70.0 ± 2.3 μm; HBK: 58.8 ±2 .3 μm; n = 4-5, p =0. 01). Thus, liver circadian clock disruption in older adult mice increases aortic stiffness with aortic apoptosis and reduced wall thickness, which may result in cardiovascular disease.