Cardioprotective Properties of Natural Anti-Platelet Activating Factor Extract From Winery By-products in Healthy Women

Study Overview
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Brief Summary
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The purpose of this study is to investigate the anti-platelet and anti-inflammatory properties of a winery by-products extract as well as to detect extract compounds and their metabolites in biological fluids. The study is a randomized, double-blind, crossover, placebo controlled postprandial study in healthy women.

Detailed Description
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The well-known cardioprotective and metabolic effects of wine consumption are mainly attributed to its micro-constituents. Grape pomace (GP) is a by-product of the winemaking process and consists mainly of skins and seeds. Winery by-products are a cheap and rich source of similar-to wine micro-constituents, which can be used either to enrich other foods or to be included in food supplements targeting the prevention or partially the therapy of cardiovascular diseases.In this line, our previous results revealed the potent in vitro anti-platelet effects of a specific ethanol-water extract rich in Platelet-Activating Factor inhibitors from winery by-products. The purpose of this study is to investigate the in vivo anti-platelet and anti-inflammatory properties of the specific winery by-products extract as well as to detect the extract compounds and their metabolites in biological fluids. Therefore a randomized double-blind, crossover, placebo controlled postprandial study in healthy women will be implemented. For this purpose, 15 healthy women will participate in the protocol. The two daily trials will take place during specific days based on their menstrual cycle. Three days before each blood collection the volunteers will be instructed to abstain from food and beverages rich in phenolic compounds and their dietary intake will be recorded (three 24h recalls and one food frequency questionnaire). The blood collections will be carried out after 8h fasting. At trial day, the volunteers will bring the first morning urine sample and anthropometric measurements will take place (weight, height, waist/hip circumference, bioelectrical impedance). Then a venous catheter will be placed and after 10 minutes the fasting blood will be collected. Volunteers will proceed to the consumption of a standardized meal (1131 kcal, 19.7% carbohydrates, 11.2% protein, 66.7% fat) along with the capsules (study extract or placebo). The type of the capsules consumed (study extract or placebo) will be randomized and blind during the two intervention days for both volunteers and investigators. Blood will be drown after the meal consumption and for the next 6h (every 30minutes for the first 4h and every 1h for the next 2h). Serum, plasma, platelet-rich plasma, leukocyte-rich plasma and urine samples will be isolated at certain time points during trial days so that the anti-platelet, anti-inflammatory and antioxidant effects of the study extract can be evaluated.

Official Title
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Investigation of in Vivo Anti-inflammatory and Anti-platelet Mechanisms of Natural Extract by Modulating PAF Metabolism and Actions

Conditions
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Platelet Aggregation and Inflammation

Intervention / Treatment
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* Behavioral: Supplement
* Behavioral: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy Females BMI: 22-32 kg/m2 Exclusion Criteria: Systematic medication Chronic disease conditions Specific dietary conditions (vegeterian, vegan...) Eating disorders

Ages Eligible for Study
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Minimum Age: 25 Years
Maximum Age: 45 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Supplement<br>Ethanol-water extract of winery by-products | Behavioral: Supplement<br>* A sufficient quantity of the winery by-products will be extracted on an industrial scale and capsules will be produced, each one containing 110mg of phenolic compounds (gallic acid equivalents).Volunteers will consume 6 capsules along with a standardized meal (1131 kcal, 19.7% carbohydrates, 11.2% protein, 66.7% fat) and blood will be drown before the meal consumption and at specific time points for the next 6h<br>|
| Placebo Comparator: Placebo<br>Maltodextrin-based placebo | Behavioral: Placebo<br>* A look-alike placebo containing maltodextrin will be prepared.Volunteers will consume 6 capsules along with a standardized meal (1131 kcal, 19.7% carbohydrates, 11.2% protein, 66.7% fat) and blood will be drown before the meal consumption and at specific time points for the next 6h<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Effect on platelet aggregation | % Change of EC50 value of platelet aggregation against PAF | Change between timepoints (before meal consumption, 30min, 90min, 150min, 210min, 300min) of the 6hour trial and between the two different interventions | 
| Effect on platelet aggregation | % Change of EC50 value of platelet aggregation against ADP | Change between timepoints (before meal consumption, 30min, 90min, 150min, 210min, 300min) of the 6hour trial and between the two different interventions | 
| Effect on platelet aggregation | % Change of EC50 value of platelet aggregation against Collagen | Change between timepoints (before meal consumption, 30min, 90min, 150min, 210min, 300min) of the 6hour trial and between the two different interventions | 
| Effect on inflammatory markers | Change in PAF biosynthetic enzymes activity (lyso-PAF AT) | Change between timepoints (before meal consumption, 60min, 120min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on inflammatory markers | Change in PAF biosynthetic enzymes activity (PAF-CPT) | Change between timepoints (before meal consumption, 60min, 120min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on inflammatory markers | Change in PAF levels Change in PAF degradation enzyme activity (Lp-PLA2) Change in PAF levels IL6 | Change between timepoints (before meal consumption, 60min, 120min, 180min, 240min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on inflammatory markers | Change in PAF degradation enzyme activity (Lp-PLA2) | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on inflammatory markers | IL-6 | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Effect on classical biochemical markers | Total serum cholesterol | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on classical biochemical markers | LDL-cholesterol | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on classical biochemical markers | HDL-cholesterol | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on classical biochemical markers | TAG | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on classical biochemical markers | uric acid | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on classical biochemical markers | Glucose | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Effect on classical biochemical markers | Insulin | Change between timepoints (before meal consumption, 0min, 30min, 60min, 90min, 120min, 150min, 180min, 210min, 240min, 300min, 360min) of the 6hour trial and between the two different interventions | 
| Detection and estimation of extract compounds metabolites |  | Change between timepoints (before meal consumption, 60min, 120min, 240min, 360min) of the 6hour trial and between the two different interventions | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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winery by-products, natural extract, postprandial study, platelet aggregation, anti-inflammatory, healthy women, Platelet-activating factor