Alteration of Mitochondrial Membrane, Fatty Acid Deficiency, Type2 Diabetes
Conditions
Keywords
Transferrin Receptor, Type 2 Diabetes, Stearic Acid (C18:0), Mitochondrial Morphology, Crossover Study
Brief summary
The purpose of this crossover study is to determine whether nutritional supplementation of C18:0 in humans has mitochondrial effects as shown in Drosophila and human cell culture. We will compare a study cohort of patients with diagnosed type 2 diabetes with non-diabetics. Participants will undergo a 2-day low-fat vegan diet and will then be supplemented with a bolus of C18:0. Changes in the mitochondrial morphology and function of white blood cells will be scored by immunofluorescence and FACS analysis.
Detailed description
The purpose of this study is to determine whether nutritional supplementation of C18:0 in humans has mitochondrial effects as shown in Drosophila and human cell culture. We will compare a study cohort of patients with diagnosed type 2 diabetes with non-diabetics. Participants will undergo a 2-day low-fat vegan diet to reach baseline levels of C18:0 and will then be fed a milkshake supplemented with 24g of C18:0, which corresponds roughly to the C18:0 content of a fast-food meal. Blood samples will be taken at baseline and several hours after intake. We will look at changes in mitochondrial morphology of neutrophils by immunofluorescence, and score mitochondrial function via FACS analysis. Since this study is designed as a crossover study, participants will also receive a mock milk shake after another 2 days of low-fat vegan diet.
Interventions
DIETARY_SUPPLEMENTC18:0
Receives 24g of C18:0 in a low-fat banana milkshake.
DIETARY_SUPPLEMENTmock
Low fat banana milkshake without C18:0 supplement.
Sponsors
German Cancer Research Center
University Hospital Heidelberg
Study design
Allocation
RANDOMIZED
Intervention model
CROSSOVER
Primary purpose
BASIC_SCIENCE
Masking
DOUBLE (Subject, Investigator)
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
Yes
Inclusion criteria
* type 2 diabetes, either dietary treatment or oral medication * must be able to give consent
Exclusion criteria
* insulin treated diabetes mellitus * severe diseases inducing wasting (e.g. cancer, liver cirrhosis, renal failure) * conditions of malnourishment * severe anemia * pregnancy * alcohol abuse
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Changes in Mitochondrial Morphology | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | Mitochondria of neutrophils are stained and scored via immunofluorescence microcsopy, either as fragmented, intermediate or fused. Statistical calculations will be performed on changes in fragmentation status after treatment. |
| Changes in Mitochondrial Function | on the day of supplementation at 0, 3 and 6 h | Mitochondrial membrane potential and ROS production in neutrophils will be analyzed via FACS. Statistical calculations will be performed on changes in the respective levels after treatment. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| plasma fatty acid levels | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | Fatty acids along with other lipid parameters like triglycerides and cholesterol for normalization purposes will be measured. |
| plasma iron, transferrin, ferritin, ferroportin and hepcidin levels | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | Measurement of iron, transferrin, ferritin, hepcidin and ferroportin from serum at all timepoints via ELISA. Changes in plasma levels will be correlated to primary endpoints. |
| diabetic late complications | 2 days before supplementation | Patients with confirmed HbA1c \> 6,5% will be considered diabetic. Then albumin in urine will be measured and diabetic neuropathy will be assessed clinically via NDS/NSS scoring. |
| insulin resistance | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | Insulin and glucose levels will be measured at each time point, HOMA index will be calculated. |
| plasma methylglyoxal levels | 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h | Methylglyoxal levels in plasma analyzed via liquid chromatography-mass spectrometry. Changes will be correlated to primary endpoints. |
Countries
Germany
Outcome results
None listed