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.
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.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
DOUBLE
Enrollment
23
University of Heidelberg
Heidelberg, Baden-Wurttemberg, Germany
Changes in Mitochondrial Morphology
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.
Time frame: 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h
Changes in Mitochondrial Function
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.
Time frame: on the day of supplementation at 0, 3 and 6 h
plasma iron, transferrin, ferritin, ferroportin and hepcidin levels
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.
Time frame: 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h
plasma methylglyoxal levels
Methylglyoxal levels in plasma analyzed via liquid chromatography-mass spectrometry. Changes will be correlated to primary endpoints.
Time frame: 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h
plasma fatty acid levels
Fatty acids along with other lipid parameters like triglycerides and cholesterol for normalization purposes will be measured.
Time frame: 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h
insulin resistance
Insulin and glucose levels will be measured at each time point, HOMA index will be calculated.
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Time frame: 2 days before supplementation, on the day of supplementation at 0, 3 and 6 h
diabetic late complications
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.
Time frame: 2 days before supplementation