Effect of a Specific Aquaporin-1 Inhibitor on Vascular Oxidative Stress in Healthy Volunteers

Study Overview
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Brief Summary
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Bacoxy_II study aims to evaluate the efficacy of a standardized Bacopa monnieri extract, KeenMind®, on vascular oxidative stress.

Detailed Description
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The food supplement Bacopa monnieri is a plant used in Ayuverda medicine, especially in the treatment of chronic neurological disease with cognitive impairment and memory disorders and for stress management. Bacopa monnieri contains several Bacosides including Bacopaside II, a specific inhibitor of aquaporin 1 (AQP1), the main water chanel found in mammalian cardiovacular tissues. AQP1, more than a water chanel, is a peroxiporin able to facilitate the passage of H2O2. AQP1 is present in myocyte, endothelial and red blood cells. Concerning the endothelial function, analyses in the FATH laboratory (IREC - UCLouvain), confirm the attenuation of H2O2 transport by AQP1 through Bacopaside II in red blood cells but also in endothelial cells. As H2O2 is involved in oxidative stress mechanisms and endothelial dysfunction, the investigators hypothezised that the oral intake of Bacopa monnieri containing the Bacopaside II could induce an inhibition of AQP1 and attenuate the passage of H2O2 leading to an attenuation of vascular oxidative stress and endothelial function. In order to answer to this question, the investigators set up the Bacoxy_II clinical study, that is a double-blind, prospective, interventional and controlled study. The study will last 4 months, including 3 months of treatment and 1 month of post-treatment follow-up. This study will include 2 groups of 20 volunteers: one group will receive a dose of 320 mg/d of Bacopa monnieri and the other group will receive the placebo treatment.

Official Title
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Randomized Controlled and Double-blinded Study of the Effect of a Specific Aquaporin-1 Inhibitor, Bacopaside II (KeenMind®), on Vascular Oxidative Stress in Healthy Volunteers

Conditions
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Cardiovascular Diseases, Oxidative Stress, Bacopaside II, Bacopa Monnieri, Endothelial Dysfunction, Aquaporin 1, AQP1

Intervention / Treatment
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* Other: KeenMind
* Other: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: - healthy volunteers Exclusion Criteria: any chronic disease use of chronic drugs or food supplements smoking

Ages Eligible for Study
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Minimum Age: 25 Years
Maximum Age: 55 Years

Sexes Eligible for Study
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All

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: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: 2 arms : placebo vs verum
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: KeenMind<br> | Other: KeenMind<br>* 320 mg/d (2 capsules/d) during 3 months<br>* Other names: Bacopa monnieri;|
| Placebo Comparator: Placebo<br> | Other: Placebo<br>* 320 mg/d (2 capsules/d) during 3 months<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Ex vivo DCFDA test on red blood cells (RBCs) | DCFA is a probe used to assess the presence of intracellular ROS. Red blood cells are incubated with DCFA and H2O2. In cells, DCFA is transformed into DCF in the presence of ROS such as H2O2 and the intensity of the emitted signal can be measured by FACS. This technique allows us to measure kinetically the entry of H2O2 by AQP1 in RBCs | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Nitrosylated hemoglobin (HbNO) | Oxidative stress is involved in the decreased bioavailability of nitric oxide (NO). HbNO a complexe used to assess NO bioavailability. HbNO can be quantify by electron paramagnetic resonance spectroscopy. | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Lipid peroxydes | Lipid peroxidation is oxidative damage that affects cellular membranes, lipoproteins, and other molecules that contain lipids in conditions with oxidative stress. Assessement of changes in lipid peroxides level during the study is a reflect of oxidative status. Lipid peroxydes are measured by an ELISA test. | Baseline (V0), 3 months (V1), 4 months (V2) | 
| EndoPAT | Endo-PAT is a non-invasive technique to assess peripheral arterial tone following a reactive hyperemia phenomenon. Using probes placed on the fingertips, EndoPAT measures changes in vascular tone mediated by the endothelium following reactive hyperemia induced by forearm occlusion using a cuff. | Baseline (V0), 3 months (V1), 4 months (V2) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| C-reactive protein (CRP) | C-reactive protein is a marker of inflammation. unit: mg/L The CRP assesses the inflammatory state of an individual. CRP is measured in the serum after a blood sampling. | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Blood count | The hemogram is a quantitative and qualitative analysis of blood constituents. This test is performed to reiterate the known safety of Bacopa monnieri on the systemic circulation after oral ingestion. The haemogram includes the following measurements: haemoglobin (g/L), haematocrit (g/L) and red blood cells (10^6/µL). | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Ion count | As Bacopa monnieri has a diuretic effect, an ionic count will help assess the concentration of electrolytes in the blood. The ion count includes the following measurements: Sodium (mM), potassium (mM), chloride (mM), bicarbonate (mM). | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Lipid count | To assess the effect of Bacopa monnieri on the lipid profile, we measure blood lipids. Lipids include total cholesterol (mg/dL), HDL cholesterol (mg/dL), LDL cholesterol (mg/dL) and triglycerides (mg/dL). | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Liver function | Liver function will be assessed to verify the known safety of Bacopa monnieri. Liver function includes: transaminase level (U/L) and gamma-glutamyl-transferase level (U/L). | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Kidney function | Renal function will be assessed to reiterate the known safety of Bacopa monnieri. Renal function includes the following measurements: creatinine (mg/dl), urea (mg/dl) and glomerular filtration rate (ml/min/m²). | Baseline (V0), 3 months (V1), 4 months (V2) | 
| HOMA index | The HOMA index will be used to assess changes in insulin sensitivity over the course of the study. The HOMA index is calculated as follows: Fasting blood glucose (mmol/L) * Fasting insulin (mui/mL)/22.5. To calculate this index, we measure fasting blood glucose (mg/dL) and fasting insulin (pmol/L). An index >2.4 is a diagnostic of insulin resistance. | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Body mass index, fat mass, lean mass, | To assess changes in body composition over the course of the study, we will be using impedance measurement technology. Impedance analysis is used to calculate lean mass (%), fat mass (%) and fluid volume (%). | Baseline (V0), 3 months (V1), 4 months (V2) | 
| Waist and hip circumference ratio | To assess changes in body composition over the course of the study, we measure waist and hip circumferences. We measure these circumferences using a tape measure in centimetres. | Baseline (V0), 3 months (V1), 4 months (V2) | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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hydrogen peroxide, cardiovascular diseases, Bacopa monnieri, Bacopaside II, endothelial dysfunction, vascular oxidative stress, aquaporin 1, AQP1, peroxiporin, H2O2