Exercise Intervention Study
Objective: To identify pathways compromised in insulin resistance and test the effect of moderate exercise on whole-body and cellular insulin action in type 2 diabetic patients, obese nondiabetics, and lean controls
Gather these items before starting the experiment. Check off items as you prepare.
Equipment1
Not specified • Not specified • Not specified • N/A
Materials1
N/A • N/A • N/A • N/A
1 item available from ConductScience
Estimated: $46.00
As an Amazon Associate, we earn from qualifying purchases. Product links help support this free resource.
Protocol Steps
Subject recruitment and stratification
Recruit and stratify subjects into three groups: type 2 diabetic patients, obese nondiabetics, and lean controls
Note: Three distinct subject populations were studied
View evidence from paper
“We conducted euglycemic clamps and muscle biopsies on type 2 diabetic patients, obese nondiabetics and lean controls”
Exercise intervention
Administer a single bout of moderate exercise to subjects
Note: Exercise was performed 24 hours before the euglycemic clamp measurement
View evidence from paper
“Exercise 24 hours before the euglycemic clamp increased phosphorylation of insulin receptor and IRS-1”
Euglycemic clamp procedure
Conduct euglycemic clamp to measure whole-body insulin action and glucose disposal 24 hours after exercise intervention
Note: Performed with and without prior exercise bout
View evidence from paper
“We conducted euglycemic clamps and muscle biopsies on type 2 diabetic patients, obese nondiabetics and lean controls, with and without a single bout of exercise”
Muscle biopsy collection
Obtain muscle tissue samples during or after euglycemic clamp for cellular analysis
Note: Biopsies were collected in conjunction with euglycemic clamp procedures
View evidence from paper
“We conducted euglycemic clamps and muscle biopsies on type 2 diabetic patients, obese nondiabetics and lean controls”
Measure phosphorylation of insulin receptor and IRS-1
Analyze muscle tissue samples for phosphorylation of insulin receptor and IRS-1 protein
Note: Measured in response to insulin stimulation
View evidence from paper
“Insulin stimulation of the phosphatidylinositol 3-kinase (PI 3-kinase) pathway, as measured by phosphorylation of the insulin receptor and IRS-1”
Measure IRS protein association with p85 and PI 3-kinase
Analyze muscle tissue for IRS protein association with p85 and PI 3-kinase regulatory subunit
Note: Association measured as indicator of PI 3-kinase pathway activation
View evidence from paper
“by IRS protein association with p85 and with PI 3-kinase”
Measure MAP kinase pathway activation
Assess insulin stimulation of the MAP kinase pathway in muscle tissue samples
Note: Compared between subject groups to determine differential pathway effects
View evidence from paper
“Insulin stimulation of the MAP kinase pathway was normal in obese and diabetic subjects”
Correlate glucose disposal with signaling markers
Analyze correlation between insulin-stimulated glucose disposal and association of p85 with IRS-1
Note: Identifies key steps in insulin resistance mechanism
View evidence from paper
“Insulin stimulation of glucose-disposal correlated with association of p85 with IRS-1”