Fasting Therapy Contributes to the Improvement of Endothelial Function and Decline in Vascular Injury‐Related Markers in Overweight and Obese Individuals via Activating Autophagy of Endothelial Progenitor Cells

Abstract

Background. High body mass index- (BMI-) related vascular injury contributes to the pathogenesis of the atherosclerotic cardiovascular disease (ASCVD). Rigorous calorie restriction is one of the major lifestyle interventions to reduce vascular risk in overweight or obese individuals. However, the effects of fasting therapy (FT) on vascular function and the mechanism are still unclear. This study was aimed to investigate the impacts of FT on endothelial function, arterial stiffness, and circulating arterial damage parameters in overweight and obese individuals and possible mechanism. Methods. Overweight and obese individuals participated in FT intervention (7-day very low calorie diet). Arterial function including brachial arterial flow-mediated dilation (FMD), brachial-ankle pulse wave velocity (baPWV), vascular injury-related markers including trimethylamine N-oxide (TMAO), and leptin and endothelial microparticles (EMPs) were assessed. Endothelial progenitor cells (EPCs) of these participants were isolated and cultured to investigate EPCs function. mRFP-GFP-LC3 confocal microscopy scanning and western blot were carried out to determine autophagy. Results. After FT, body weight and BMI significantly decreased (81.76 ± 12.04 vs. 77.51 ± 12.06 kg, P<0.01; 29.93 ± 2.82 vs. 28.47 ± 2.83 kg/m2, P<0.01). FT remarkably improved FMD (5.26 ± 1.34 vs. 6.25 ± 1.60%, P=0.01) while baPWV kept unchanged. TMAO and leptin levels decreased (3.96 ± 1.85 vs. 2.73 ± 1.33 μmol/L, P=0.044; 6814 ± 2639 vs. 3563 ± 2668 μmol/L, P<0.01). EMPs showed a decreased tendency. EPCs function was significantly improved, autophagy fluorescence intensity was enhanced, and the level of Beclin1, Atg5, LC3 II/I also increased after starvation in vitro, and the effects were blocked by autophagy inhibitor. Conclusions. Our present study demonstrated for the first time that FT markedly improves endothelial function and reduces the levels of arterial injury markers through improving EPCs function via activating autophagy. These findings provide a novel insight into FT as a lifestyle intervention strategy to promote the maintenance of vascular homeostasis in overweight or obese individuals. The trial was registered with ChiCTR1900024290.

Association of enhanced circulating trimethylamine N‐oxide with vascular endothelial dysfunction in periodontitis patients

“…Western blot analysis of EPCs was performed as described previously 23 . The following antibodies were used: anti‐N‐terminal GSDME antibody, anti‐cleaved caspase 3 antibody, and anti‐β actin antibody.…”

“…The plasma TMAO was assessed using high‐performance liquid chromatography with tandem mass spectrometry (HPLC‐MS/MS) 23 . Briefly, 10 μL d9‐TMAO was added to 100 uL plasma; then 300 μL of acetonitrile was added for protein precipitation, vortexed for 1 min, centrifugated at 1000 rpm, 4°C for 5 minutes; finally, we took 200 µL of the remaining supernatant for analysis.…”

“…Western blot analysis of EPCs was performed as described previously. 23 The following antibodies were used: anti-Nterminal GSDME antibody † † † , anti-cleaved caspase 3 antibody ‡ ‡ ‡ , and anti-β actin antibody § § § . Proteins were visualized with HRP-conjugated anti-rabbit IgG **** , followed by the use of the ECL chemiluminescence system † † † † .…”

Religious fasting and the vascular health

“…Numerous health benefits of fasting have been supported by experimental 15 and clinical studies. 16 , 17 Decreased inflammation, neuroendocrine activation, reduced mitochondrial oxidative stress, 11 and activation of autophagy in endothelial cells that accompany intermittent fasting prevent cell death 18 and promote proliferation, regeneration, and activation of endothelial progenitor cells. 18 Fasting has beneficial effects on cardiovascular health through modification of common risk factors such as better blood pressure control, 11 increased insulin sensitivity and better control of blood glucose level.…”

Fasting-Mimicking Diet Reduces Trimethylamine N-Oxide Levels and Improves Serum Biochemical Parameters in Healthy Volunteers

“…Although there is some evidence that TMAO levels could be targeted by some types of caloric restriction [ 30 , 31 , 32 ], reduced protein intake [ 33 ] or diets supplemented with sources of dietary fibers and unsaturated fatty acids [ 34 , 35 ], it has not been thoroughly investigated. As FMD has shown potential in reducing CVD risks, our objective is to investigate whether this short-term reduction in caloric intake by decreasing the consumption of animal-derived proteins, as in the case of FMD, could also serve as an effective dietary strategy to reduce plasma TMAO levels in healthy omnivorous volunteers.…”