Consequences of Hypoglycaemia on Cardiovascular and Inflammatory Responses

Not Applicable

Completed

Conditions: Inflammatory Response
Hypoglycemia
Diabetes Mellitus, Type 1
Diabetes Mellitus, Type 2

Interventions: Procedure: hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp

Brief Summary: People with Type 1 diabetes (T1DM), type 2 diabetes (T2DM) and healthy volunteers will undergo a hypoglycaemic clamp to to investigate the effect of hypoglycaemia on cardiovascular and inflammatory responses.

Detailed Description: Objectives: The overall aim of the present study is to investigate the effect of hypoglycaemia on cardiovascular and inflammatory responses, molecular mechanisms and epigenetic profiles in various groups of people with diabetes type 1, type 2 and healthy volunteers.

Study design: Intervention study

Intervention: All subjects will undergo a hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp (nadir 2.8 mmol/L), during and after which blood and urine will be sampled for further examination for up to one week.

Recruitment & Eligibility

Inclusion Criteria

Overall inclusion criteria

Ability to provide written informed consent
Must be able to speak and read Danish (for Hillerød-site) and Dutch (for Nijmegen-site)
Insulin treatment according to basal-bolus insulin regimen (injections or insulin pump) (except for group 5)
Body-Mass Index: 19-40 kg/m2
Age ≥18 years, ≤ 80 years
Blood pressure: <140/90 mmHg
Duration of diabetes > 1 year (except for group 5)
HbA1c < 100 mmol/mol

Group specific

Group 1: HbA1c >64 mmol/mol
Group 2: impaired awareness of hypoglycaemia (IAH) as assessed by a score of ≥3 on the modified Clarke questionnaire, ≥4 on the Gold questionnaire and a positive score on the Pedersen-Bjergaard questionnaire.
Group 3: normal awareness of hypoglycaemia (NAH) as assessed by a score of <3 on the modified Clarke questionnaire, <4 on the Gold questionnaire and a negative score on the Pedersen-Bjergaard.
Group 4: Insulin treatment for at least 1 year
Group 5/6: HbA1c <42 mmol/mol

Exclusion Criteria

- Severe medical or psychological conditions interfering with the perception of hypoglycaemia other than IAH such as brain injuries, epilepsy, a major cardiovascular disease event or anxiety disorders
Use of immune-modifying drugs or antibiotics
Treatment with glucose-modifying (other than insulin, SGLT-2 inhibitors and metformin) agents (e.g. prednisolon)
Use of anti-depressive drugs
Pregnancy or breastfeeding or unwillingness to undertake measures for birth control
Use of statins (e.g. stop statins >2 weeks before performing blood sampling. This can be safely done in the context of primary prevention)
Any event of cardiovascular disease in the past 5 years (e.g. myocardial infarction, stroke, heart failure, symptomatic peripheral arterial disease)
Auto-inflammatory or auto-immune diseases
Any infection in past three months
Previous vaccination in the past three months
Laser coagulation for proliferative retinopathy in the past six months
Proliferative retinopathy
Diabetic nephropathy as reflected by an albumin-creatinine ratio ˃ 30 mg/gor an estimated glomerular filtration rate (by MDRD) ˂60ml/min/1.73m2
History of pancreatitis (acute or chronic) or pancreatic cancer

Arm && Interventions

Group	Intervention	Description
T1DM poor glycaemic control	hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp	patients with type 1 diabetes and poor glycaemic control (HbA1c \>8% / \>64 mmol/mol will undergo hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp
T1DM Normal awareness	hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp	patients with type 1 diabetes and normal awareness of hypoglycaemia will undergo hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp
Healthy control T1DM	hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp	Healthy controls without diabetes and age, gender and BMI matched with diabetes type 1 participants will undergo hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp
T1DM impaired awareness	hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp	patients with type 1 diabetes and impaired awareness of hypoglycaemia will undergo hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp
T2DM + Insulin	hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp	patients with type 2 diabetes with insulin treatment for at least 1 year will undergo hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp
Healthy control T2DM	hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp	healthy controls without diabetes and age, gender and BMI matched with diabetes type 2 participants will undergo hyperinsulinemic normoglycaemic-hypoglycaemic glucose clamp

Primary Outcome Measures

Name	Time	Method
Inflammatory responses of hypoglycaemia by measuring the cytokine production of isolated monocytes using ELISA	1.5 year	Cytokine production (TNF-alfa, IL-6, IL-10 and IL-1β) of isolated and stimulated monocytes

Secondary Outcome Measures

Name	Time	Method
Metabolomics profile of each group	1.5 year	Un-targeted metabolomics and identification of metabolites based on exact mass using metabolomics library
Cardiac function responses to hypoglycaemia using echocardiography	1.5 year	Cardiac function responses to hypoglycaemia using echocardiography
Oxidative stress responses using oxidative stress marker	1.5 year	Excretion of guanine nucleosides in urine (ng/mL)
Epigenetic modifications	1.5 year	Epigenetic modifications due to hypoglycaemia in the promoter regions of the pro-inflammatory cytokines in monocytes
Atherogenic responses of (recurrent) hypoglycaemia using foam cell formation.	1.5 year	Measurement of Ox-LDL uptake by measuring intracellular apolipoproteine B
Cognitive function responses to hypoglycaemia using cognitive function tests (TAP, PASAT)	1.5 year	Amount of correct answers

Locations (2): Nordsjællands University Hospital
🇩🇰
Hillerød, Nordsjaelland, Denmark
Radboudumc
🇳🇱
Nijmegen, Gelderland, Netherlands