Our recent data in mice have demonstrated a key role of xanthine oxidase in hyperglycemia-induced by Reactive oxygen species production, and a preventive role of allopurinol (inhibitor of xanthine oxidase) on the keeping of mitochondria number and structure, in skeletal muscle of diabetic mice. The investigators want to initiate a clinical trial in order to evaluate the efficacy of allopurinol on the improvement of mitochondrial alterations, oxidative capacities and insulin sensitivity, in skeletal muscle of type 2 diabetic patients.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
DOUBLE
Enrollment
31
2 capsules of allopurinol 150 mg daily for 3 month
2 capsules of lactose daily for 3 month
CRNH Rhône Alpes
Lyon, France
muscle oxidative stress in diabetic patients
muscular protein carbonylation level (unit: reactive carbonyl derivates, arbitrary unit) by Western blot (oxyblot kit from Chemicon) and pro et antioxidant genes expression (unit: mRNA levels normalized by housekeeping gene, arbitrary ratio) by real time polymerase chain reaction (RT-PCR)
Time frame: At 3 months of treatment
- plasmatic oxidative stress by dosing plasmaticmarkers:
Dosing plasmatic malondialdéhyde, plasmatic H2O2, protein carbonylation of plasmatic protein and urinary isoprostans, and finally antioxidants (vitamins C and E, glutathione (unit: from µM to M)
Time frame: At 3 months of treatment
alterations in mitochondrial structure of skeletal muscle with transmission electron microscopy
Analysis of mitochondria area (in µm2) and density (in %)
Time frame: At 3 months of treatment
mitochondrial density by measuring the ratio mitochondrial Deoxyribonucleic acid (mtDNA)/nuclear DNA by real-time polymerase chain reaction (PCR) in skeletal muscle
Time frame: At 3 months of treatment
mitochondrial function
Expression of genes implicated in mitochondrial action (messenger Ribonucleic acid (mRNA) levels by Reverse transcription polymerase chain reaction (RT-PCR) and proteins levels by Western Blot)(unit: arbitrary ratio relative to housekeeping gene/protein).
Time frame: At 3 months of treatment
quantification of intramuscular lipids by histology (biopsy analysis)
Staining Oil Red O evaluate the intramuscular lipid accumulation using the software ImageJ(unit: % of labelling by field).
Time frame: At 3 months of treatment
sensitivity to insulin using a hyperinsulinemic euglycemic clamp
sensitivity to insulin will be expressed as the glucose infusion rate (GIR)/insulinemia ratio.
Time frame: At 3 months of treatment
uricemia and xanthine oxidase activity in sera and muscles(unit: mg/l for uricemia and mU/ml for XO activity)
Plasma concentrations of uric acid will be measured before and after treatment to assess patient compliance . The reduction of xanthine oxidase activity in serum and muscle protein lysates will be measured using the kit " Amplex Red xanthine / xanthine oxidase assay kit" from Molecular Probes
Time frame: At 3 months of treatment
Tolerance of the treatment measured by any adverse events during treatment and between each visit.
Any adverse events during treatment and between each visit.
Time frame: during the 3 months of treatment
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