Clinical Trial: A Study of Intracellular Signaling in Muscle and Fat Cells During Ketosis

Study Status: Completed
Recruit Status: Completed
Study Type: Interventional

Official Title: The Role of ATGL and G0/G1 Switch Gene Complex in Lipopolysaccaride (LPS) Induced Ketosis - a Controlled, Randomised, Clinical Experimental Study

Brief Summary:

Hypothesis

  1. To define whether stimulation of ATGL and suppression of G0/G1 switch gene occur in the initial phases of diabetic ketoacidosis and thus can be identified as the primary mechanisms behind this life threatening condition.
  2. Make a human model for studying ketoacidosis.

The investigators plan to reduce in their regular insulin over night. In the morning we administer endotoxin, which together with a relative lack of insulin will initiate ketogenesis - a state of ketoacidosis. On another occasion strict glycemic control is imposed by means of intravenous insulin. The testing is done two separate days with at least 3 weeks in between and patients are admitted to hospital the evening before the day of testing. The investigators use isotopic tracers to determine metabolic fluxes and analyse fat (ATGL, G0/G1 switch gene) and muscle biopsies.


Detailed Summary:
Sponsor: University of Aarhus

Current Primary Outcome: Insulin signaling expressed as a CHANGE in phosphorylation of intracellular target proteins and CHANGE in mRNA expression of target genes in muscle- and fat-tissue. [ Time Frame: Muscle and fat biopsies obtained on each study day (arm): t1= 6.45 (-75min) am t2=11.15 (195min) am t3= 12.30 pm (270min) ]

Change in phosphorylation of target proteins and messenger RNA (mRNA) expression of target genes assessed with western blotting technique.


Original Primary Outcome: Same as current

Current Secondary Outcome:

  • Change in Intracellular markers of lipid metabolism in muscle- and fat tissue biopsies [ Time Frame: Muscle and fat biopsies obtained on each study day (arm): t1= 6.45 am (-75min) t2=11.15 (195min) am t3= 12.30 pm (270min) ]
    Muscle and fat at t1 and t2. Muscle biopsy at t3. Intracellular markers are assessed by western blotting.
  • Metabolism [ Time Frame: Change in glucose, fat and protein metabolism between study days and during each study day ]
    Change in glucose, fat and protein metabolism assessed by tracer kinetics on every study day (specific times below) and by indirect calorimetry. [3H 3]Glucose tracer from t=0 - 360min. Palmitic acid tracer from t=165min - 360min. Urea tracer from 0min - 240min. amino acid tracer from 60 min - 360 min.
  • Cytokines and stress hormones [ Time Frame: In basal period t=0-240 minutes and in clamp period t=240-390 minutes ]
    Measurement of immune response to endotoxin and hypoinsulinaemia. Estimating the whole body stress during ketoacidosis and pre ketoacidosis.


Original Secondary Outcome: Same as current

Information By: University of Aarhus

Dates:
Date Received: May 23, 2014
Date Started: June 2014
Date Completion:
Last Updated: December 1, 2015
Last Verified: June 2014