Plant Sterols on Cardiovascular Markers, Microbiota and Sterol Metabolism (Cardiofoodsterol)

Effect of Plant Sterols on Inflammatory, Endothelial Function and Oxidative Stress Markers, Microbiota and Sterol Metabolism in a Cardiovascular Risk Population

Potential cholesterol-lowering effect of a regular intake of a plant sterol (PS)-containing food supplement, in overweight/obese type 1 or 2, normoglycemic/pre-diabetic, with LDL-cholesterol values > 115 mg/dl and not pharmacologically treated participants treated with the PS-containing food supplement or placebo supplement.

Study Overview

• Status: Recruiting
• Conditions: Cardiovascular Diseases; Prediabetic State; Hypercholesterolemia; Overweight and Obesity
• Intervention / Treatment: Dietary supplement: Plant sterols 2g/day; Dietary supplement: Placebo

Detailed Description

Cardiovascular disease (CVD) is the leading cause of death worldwide, with hypercholesterolemia being one of the main risk factors for CVD. The deposition and oxidation of LDL-cholesterol particles triggers a series of molecular events favoring chronic low-grade inflammation, endothelial dysfunction and oxidative stress. This situation promotes atherogenesis thus increasing cardiovascular risk. Obesity favors the secretion of pro-inflammatory mediators and promotes the recruitment of macrophages to adipose tissue, insulin resistance, hyperglycemia and hyperlipidemia, thus increasing the risk of CVD. In addition, obesity has been associated with gut dysbiosis, which in turn is associated with atherosclerosis in some studies. Beneficial effects of PS on LDL-cholesterol and inflammatory, endothelial dysfunction and oxidative stress markers have been reported by several clinical trials. A meta-analysis suggests a lowering effect of PS on body mass index (BMI) in participants with BMI>25. Furthermore, the consumption of PS has been beneficially associated in in vitro studies with changes in intestinal microbial profile, sterol metabolism and short chain fatty acids (SCFA) production. Therefore, the hypothesis is if the consumption of PS as a food supplementation could reduce cardiovascular risk. The present study aims to evaluate the LDL-cholesterol serum levels after regular intake of a food supplement containing PS (2 g/day) in overweight/obese type 1 or 2 patients, normoglycemic /pre-diabetic and with LDL-cholesterol values > 115 mg/dl not pharmacologically treated. This is a crossover study with 21 participants (intake of a food supplement containing PS) and 21 participants (intake of excipient-based placebo), with a first intervention period of 8 weeks. After a 6-week washout period, the treatments are switched, with a second intervention period of 8 weeks. In addition, to the LDL-cholesterol lowering assessment, other biochemical, hematological, inflammatory, endothelial dysfunction and oxidative stress parameters are assessed in serum samples. Moreover, sterol and metabolite profiling in serum and feces, microbiota modulation, anthropometric measurements and body composition, bioimpedance, dietary intake and physical activity questionnaire are evaluated. All parameters are evaluated at the beginning (weeks 0 and 14) and at the end of each intervention period (weeks 8 and 22).

• Study Type: Interventional
• Enrollment (Estimated): 42
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Guadalupe Garcia Llatas, Professor; Phone Number: +34 963543766; Email: guadalupe.garcia@uv.es

Study Locations

• Spain
  • Valencia, Spain, 46026
    • Recruiting
    • Hospital Universitario y Politécnico La Fe de Valencia
    • Contact: Juan Francisco Merino Torres

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

Description

Inclusion Criteria:

• BMI: 27-29.9 or 30-39.9
• Plasmatic glucose: < 100mg/dl or 100-125mg/dl
• Glycosylated hemoglobin: < 5.7 or 5.7-6.4
• LDL cholesterol > 115mg/dL
• Serum levels of biochemical and hematological parameters and fat-soluble vitamins within reference ranges.

Exclusion Criteria:

• Subjects on cholesterol-lowering pharmacological treatment
• Smokers
• Alcohol consumption above 30 g/day
• Pregnant or lactating women
• Any infection, serious illness or co-morbidity that may affect the bioavailability of PS (e.g., malabsorption, celiac disease, allergies or food intolerances)
• Diseases of the gastrointestinal tract
• Antibiotic, hormonal or anabolic treatment
• Participants consuming foods enriched with PS or food supplements that contain PS
• Participants who follow specialist weight loss diets, vegans or vegetarians

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Basic Science
• Allocation: Randomized
• Interventional Model: Crossover Assignment
• Masking: Triple

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: Dietary Supplement; PS-containing dietary supplement Sachet containing a powdered microencapsulated free plant sterols (2 g ingredient/day)	Dietary supplement: Plant sterols 2g/day; PS-containing dietary supplement Sachet containing a powdered microencapsulated free plant sterols (2 g ingredient/day) during 8 weeks
Placebo Comparator: Placebo; Sachet containing the powdered excipients of the dietary supplement	Dietary supplement: Placebo; Sachet containing the powdered excipients of the dietary supplement during 8 weeks

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in plasmatic LDL-c	LDL-c, calculated by the Friedewald's formula, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Changes in plasmatic total cholesterol	Total cholesterol, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic HDL-c	HDL-c, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic non-HDL cholesterol	Non-HDL cholesterol, calculated by subtracting HDL-c from total cholesterol, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic triglycerides	Triglycerides, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic Apo A	Apo A, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic Apo B	Apo B, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention	0, 8, 14 and 22 weeks
Changes in plasmatic fibrinogen	Fibrinogen, assessed by Clauss coagulometric method, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic High-sensitivity C-reactive protein (hsCRP)	hs CRP, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic glucose	Glucose, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic insulin	Insulin, assessed by enzymatic-colorimetric methods, with repeated measures (at the beginning and at the end of each period the intervention	0, 8, 14 and 22 weeks
Changes in plasmatic Homeostatic Model Assessment for Insulin Resistance (HOMA-IR)	HOMA-IR, calculated by mathematical formula from the fasting insulin and fasting glucose levels, with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum levels of the sterols and metabolites profile	Profile sterols and metabolites, assessed by gas chromatography coupled with flame ionization detector (GC-FID), with repeated measures (at the beginning and at the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in feces levels of the sterols and metabolites profile	Profile sterols and metabolites, assessed by gas chromatography coupled with flame ionization detector (GC-FID), with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum interleukin 10 (IL-10)	IL-10, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum interleukin 12p70 (IL-12p70)	IL-12p70, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum interleukin 1β (IL-1β)	IL-1β, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum interleukin 6 (IL-6)	IL-6, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum interleukin 8 (IL-8)	IL-8, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum tumor necrosis factor alpha (TNFα)	TNFα, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum vascular cell adhesion molecule-1 (VCAM-1)	VCAM-1, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum E-Selectin	E- Selectin, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in serum endothelin (ET-1)	ET-1, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention	0, 8, 14 and 22 weeks
Changes in serum plasminogen activator inhibitor-1(PAI-1)	PAI-1, assessed by immunochemical techniques, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic LDL-cholesterol oxidation	LDL-cholesterol oxidation, assessed by enzyme kits, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic 8-isoprostane	8-isoprostane, assessed by enzyme kits, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic malondialdehyde (MDA)	MDA, assessed by ultra-performance liquid chromatography-MS (UPLC-MS), with repeated measures (at the beginning and the end of each period the intervention	0, 8, 14 and 22 weeks
Changes in reduced glutathiones (GSH)	GSH, assessed by ultra-performance liquid chromatography-MS (UPLC-MS), with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in oxidized glutathione (GSSG)	GSSG, assessed by ultra-performance liquid chromatography-MS (UPLC-MS), with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in the composition of microbiota in feces	Composition of microbiota, assessed by genetic sequencing and subsequent biostatistical analysis, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in feces of short-chain fatty acids (SCFAs)	SCFAs, assessed by high-performance liquid chromatography with an ultraviolet detector (HPCL-UV), with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in plasmatic levels of cholesterol oxidation products (COPs)	COPs, assessed by gas chromatography-mass spectrometry, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in body weight (WT)	WT, assessed by standard balance scale, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in body height (HT)	HT, assessed by tape measure, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in body circumferences	Circumferences, assessed by tape measure, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Changes in Bioelectrical impedance analysis (BIA)	BIA, assessed by bio-impedance meter, with repeated measures (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Assessment of dietary intake	Food frequency survey Questionnaire (FFQ) is used, consisting in 136 questions divided into the different food groups (dairy products; eggs, meat and fish; vegetables; fruit; pulses and cereals; oils and fats; bakery and confectionery; miscellaneous; drinks) indicating the average frequency of consumption during the past year, which can be scored as never or almost never, per month (1-3), per week (1, 2-4,5-6), per day (1, 2-3, 4-6, 6+), with repeated of questionnaire (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Evaluation of the physical activity to measure quality of life	International physical activity questionnaire-short form (IPAC-SF) is used, consisting in 7 questions. Intensity, frequency and duration of the exercise are evaluated through metabolic equivalent of task (METs). This allows to differentiate 3 levels of physical activity: Low: Not enough activity to achieve the next level Moderate: 3 or more days of vigorous physical activity for at least 20 minutes per day, 5 or more days of moderate physical activity and/or walking at least 30 minutes per day, or 5 or more days of any combination of walking, moderate or vigorous physical activity achieving at least a total of 600 METs. High: Vigorous physical activity at least 3 days per week achieving a total of a least 1500 METs, or 7 days of any combination of walking, with moderate and/or vigorous physical activity, achieving a total of a least 3000 METs. With repeated of questionnaire (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks
Evaluation of the Mediterranean diet adherence to measure quality of life	Mediterranean diet adherence screener (MEDAS) is used, consisting in 14 questions (each one 0 or 1 point, final score between 0 and 14). Results are ranged between 0-7 points (low adherence), 7-10 (moderate adherence), and 10-14 (high adherence) With repeated of questionnaire (at the beginning and the end of each period the intervention)	0, 8, 14 and 22 weeks

Collaborators and Investigators

• Sponsor: University of Valencia
• Collaborators: University of Bologna; Hospital Universitario La Fe
• Investigators: Principal Investigator: Guadalupe Garcia Llatas, Professor, University of Valencia

Publications and helpful links

General Publications

• Blanco-Morales V, Garcia-Llatas G, Yebra MJ, Sentandreu V, Lagarda MJ, Alegria A. Impact of a Plant Sterol- and Galactooligosaccharide-Enriched Beverage on Colonic Metabolism and Gut Microbiota Composition Using an In Vitro Dynamic Model. J Agric Food Chem. 2020 Feb 19;68(7):1884-1895. doi: 10.1021/acs.jafc.9b04796. Epub 2019 Sep 26.
• Blanco-Morales V, Silvestre RLA, Hernandez-Alvarez E, Donoso-Navarro E, Alegria A, Garcia-Llatas G. Influence of Galactooligosaccharides on the Positive Effect of Plant Sterol-Enriched Beverages on Cardiovascular Risk and Sterol Colon Metabolism. J Agric Food Chem. 2022 Jan 19;70(2):532-542. doi: 10.1021/acs.jafc.1c06120. Epub 2022 Jan 11.
• Caudet J, Trelis M, Cifre S, Tapia G, Soriano JM, Rodrigo R, Merino-Torres JF. Do Intestinal Unicellular Parasites Have a Role in the Inflammatory and Redox Status among the Severely Obese? Antioxidants (Basel). 2022 Oct 23;11(11):2090. doi: 10.3390/antiox11112090.
• Heggen E, Kirkhus B, Pedersen JI, Tonstad S. Effects of margarine enriched with plant sterol esters from rapeseed and tall oils on markers of endothelial function, inflammation and hemostasis. Scand J Clin Lab Invest. 2015 Apr;75(2):189-92. doi: 10.3109/00365513.2014.992040. Epub 2015 Jan 1.
• Oliveira Godoy Ilha A, Sutti Nunes V, Silva Afonso M, Regina Nakandakare E, da Silva Ferreira G, de Paula Assis Bombo R, Rodrigues Giorgi R, Marcondes Machado R, Carlos Rocha Quintao E, Lottenberg AM. Phytosterols Supplementation Reduces Endothelin-1 Plasma Concentration in Moderately Hypercholesterolemic Individuals Independently of Their Cholesterol-Lowering Properties. Nutrients. 2020 May 22;12(5):1507. doi: 10.3390/nu12051507.
• Mannarino E, Pirro M, Cortese C, Lupattelli G, Siepi D, Mezzetti A, Bertolini S, Parillo M, Fellin R, Pujia A, Averna M, Nicolle C, Notarbartolo A. Effects of a phytosterol-enriched dairy product on lipids, sterols and 8-isoprostane in hypercholesterolemic patients: a multicenter Italian study. Nutr Metab Cardiovasc Dis. 2009 Feb;19(2):84-90. doi: 10.1016/j.numecd.2008.03.012. Epub 2008 Aug 31.
• Menendez-Carreno M, Steenbergen H, Janssen HG. Development and validation of a comprehensive two-dimensional gas chromatography-mass spectrometry method for the analysis of phytosterol oxidation products in human plasma. Anal Bioanal Chem. 2012 Feb;402(6):2023-32. doi: 10.1007/s00216-011-5432-2. Epub 2011 Oct 5.

Study record dates

Study Major Dates

• Study Start (Actual): May 21, 2024
• Primary Completion (Estimated): December 1, 2024
• Study Completion (Estimated): February 1, 2025

Study Registration Dates

• First Submitted: June 12, 2024
• First Submitted That Met QC Criteria: June 26, 2024
• First Posted (Actual): July 1, 2024

Study Record Updates

• Last Update Posted (Actual): July 1, 2024
• Last Update Submitted That Met QC Criteria: June 26, 2024
• Last Verified: June 1, 2024

More Information

Terms related to this study

• Keywords: Inflammation; Physical Activity; Microbiota; Oxidative Stress; Endothelial function; Plant Sterols; Dietary Intake; Bioimpedance; Cholesterol LDL; Sterol Metabolites
• Additional Relevant MeSH Terms: Glucose Metabolism Disorders; Metabolic Diseases; Endocrine System Diseases; Diabetes Mellitus; Overnutrition; Nutrition Disorders; Body Weight; Lipid Metabolism Disorders; Hyperlipidemias; Dyslipidemias; Cardiovascular Diseases; Prediabetic State; Overweight; Hypercholesterolemia
• Other Study ID Numbers: BIONUTEST_2024-0358-1; PID2019-104167RB-I00 (Other Grant/Funding Number: MCIN/AEI/10.13039/501100011033)

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No