Evaluating the role of orange juice, HESPERidin in vascular HEALTH benefits (HESPER-HEALTH study): protocol for a randomised controlled trial

Abstract

Introduction: Although epidemiological studies associate the consumption of sugary beverages with adverse health effects, human experimental studies have demonstrated substantially different metabolic responses when 100% fruit juices are compared with artificial beverages. Fruit juices do not just provide sugars and associated calories, but they are also rich in bioactive compounds. Flavanones are bioactives specifically and abundantly found in citrus foods, with hesperidin as the major representative in sweet oranges. Flavanone intake has been associated with a lower incidence of mortality from cardiovascular disease (CVD). However, clinical evidence are too scarce to confirm the vasculoprotective effects of 100% orange juice (OJ) presumably mediated by flavanones and thereby do not allow firm conclusions to be drawn about their efficacy.

Methods and analysis: The HESPER-HEALTH study aims to assess the efficacy of OJ in improving vascular function and the contribution of hesperidin to these effects. This double-blind, randomised, controlled, crossover study will be carried out in 42 volunteers predisposed to CVD, based on age and on overweight. It includes three 6-week periods of consumption of 330 mL/d of OJ versus control drinks with and without hesperidin at a dose in agreement with a daily OJ serving (approx. 200-215 mg). The primary outcome is endothelial function, assessed by flow mediated dilation, with measurements performed at fasting and postprandially in response to a challenge meal. The secondary outcomes include bioavailability and metabolism of flavanones, changes in other markers of vascular function, systemic biomarkers of cardiovascular risk, endothelial dysfunction and inflammation, vitamin C and carotenoids status, anthropometry and body composition, gut microbiota composition, nutrigenomic response and in oxylipin profiling.

Ethics and dissemination: This ongoing study was approved by the Ethics committee Sud-Est III, Bron, France on 17 November 2020. The trial is registered on ClinicalTrials.gov. The results will be disseminated in peer-reviewed journals.

Trial registration number: NCT04731987; Pre-results.

Keywords: molecular biology; nutrition & dietetics; vascular medicine.

Conflict of interest statement

Competing interests: None declared.

© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

Figures

Figure 1

General scheme of HESPER-HEALTH study progress. V: visits in PIC/CIC; Drink A: OJ naturally rich in hesperidin; Drink B: control beverage with sugar concentrations identical to (A); Drink C: control beverage identical to (B) but supplemented with hesperidin at the level of (A); FR: 3-day food record. OJ, orange juice; PIC/CIC, plate-forme d'investigation clinique/centre d'investigation clinique.

Figure 2

HPLC-DAD chromatogram of an OJ from a commercially available OJ concentrate naturally rich in hesperidin (A), a control beverage with an identical total sugar concentration (B) and a control beverage additionally supplemented with hesperidin (C). Flavonoids were extracted according to IFU and analysed by HPLC-DAD using a C18 column (250×4.6 mm, particle size 5.0 µm, Kinetex, Phenomenex, Aschaffenburg, Germany) and an acetonitrile-based elution gradient. HPLC-DAD, high-pressure liquid chromatography – diode array detection; OJ, orange juice.

Figure 3

Time table of the conduct and measurements of the HESPER-HEALTH study. The crosses indicate the visits and types of samples taken (X) and the recommendations and measurements made for each of them (x). ALAT, alanine amino transferase; ASAT, aspartate amino transferase; BP, blood pressure; DBP, diastolic blood pressure; FLD, flowmetry by laser Doppler; FMD, flow mediated dilatation; Gamma GT, gamma glutamyl transferase; HDL-chol, high-density lipoprotein cholesterol; hsCRP, high sensitivity C reactive protein; ICAM, intercellular adhesion molecule; IL-6, interleukin-6; PIC/CIC, plateforme d'investigation clinique/centre d'investigation clinique; PWV, pulse wave velocity; SBP, systolic blood pressure; TAG, triacylglycerol; TNFα, tumour necrosis factor alpha; TSH, thyroid stimulating hormone; VCAM, vascular cell adhesion molecule.