Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones

Research output: Contribution to journalJournal articlepeer-review

Standard

Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones. / Pedersen, Kent; Andersen, Helle; Fledelius, Christian; Holst, Jens Juul; Hjuler, Sara Toftegaard; Kuhre, Rune Ehrenreich.

In: Molecular Metabolism, Vol. 69, 101689, 2023.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Pedersen, K, Andersen, H, Fledelius, C, Holst, JJ, Hjuler, ST & Kuhre, RE 2023, 'Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones', Molecular Metabolism, vol. 69, 101689. https://doi.org/10.1016/j.molmet.2023.101689

APA

Pedersen, K., Andersen, H., Fledelius, C., Holst, J. J., Hjuler, S. T., & Kuhre, R. E. (2023). Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones. Molecular Metabolism, 69, [101689]. https://doi.org/10.1016/j.molmet.2023.101689

Vancouver

Pedersen K, Andersen H, Fledelius C, Holst JJ, Hjuler ST, Kuhre RE. Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones. Molecular Metabolism. 2023;69. 101689. https://doi.org/10.1016/j.molmet.2023.101689

Author

Pedersen, Kent ; Andersen, Helle ; Fledelius, Christian ; Holst, Jens Juul ; Hjuler, Sara Toftegaard ; Kuhre, Rune Ehrenreich. / Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones. In: Molecular Metabolism. 2023 ; Vol. 69.

Bibtex

@article{d40d00909f2e4e448723ec774935c733,
title = "Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones",
abstract = "Objective: A fundamental difference between physiological and pharmacological studies in rats and humans is that withdrawal of blood from conscious rats necessitates restraint which inevitably inflicts a higher level of stress. We investigated the impact of handling on acute glucose regulation and secretion of glucoregulatory hormones in rats. Methods: Fasted male Sprague Dawley rats (375–400 g, n = 11) were given an oral glucose tolerance test (OGTT) by gavage (2 g/kg). Blood was sampled frequently until 90 min after challenge by handheld sampling (HS) or by automated sampling (AS). In the HS experiment, blood was withdrawn by restraint and sublingual vein puncture; two weeks later, samples were obtained by AS through an implanted catheter in a carotid artery, allowing sampling without disturbing the animals. Results: On the day of HS, post challenge glucose AUCs were ∼17% higher (P < 0.0001), despite gastric emptying (AUC) being reduced by ∼30% (P < 0.0001). Plasma insulin AUC was 3.5-fold lower (P < 0.001), and glucose-dependent insulinotropic peptide (GIP) AUC was reduced by ∼36% but glucagon-like peptide-1 concentrations were not affected. Glucagon concentrations were higher both before and after challenge (fold difference in AUCs = 3.3). Adrenocorticotropin (ACTH) and corticosterone AUCs were 2.4-fold and 3.6-fold higher (P < 0.001), respectively. Discussion and conclusion: Our study highlights that sampling of blood from conscious rats by sublingual vein puncture inflicts stress which reduces glucose absorption and glucose tolerance and blunts secretion of insulin and GIP. As blood sampling in humans are less stressful, standard procedures of conducting OGTT's in rats by HS presumably introduce an interspecies difference that may have negative consequences for translatability of test results.",
keywords = "Glucose intolerance, Handheld blood sampling, Incretin hormones, Insulin secretion, Rats, Stress hormones",
author = "Kent Pedersen and Helle Andersen and Christian Fledelius and Holst, {Jens Juul} and Hjuler, {Sara Toftegaard} and Kuhre, {Rune Ehrenreich}",
note = "Publisher Copyright: {\textcopyright} 2023 The Author(s)",
year = "2023",
doi = "10.1016/j.molmet.2023.101689",
language = "English",
volume = "69",
journal = "Molecular Metabolism",
issn = "2212-8778",
publisher = "Elsevier",

}

RIS

TY - JOUR

T1 - Standard procedures for blood withdrawal in conscious male rats induce stress and profoundly affect glucose tolerance and secretion of glucoregulatory hormones

AU - Pedersen, Kent

AU - Andersen, Helle

AU - Fledelius, Christian

AU - Holst, Jens Juul

AU - Hjuler, Sara Toftegaard

AU - Kuhre, Rune Ehrenreich

N1 - Publisher Copyright: © 2023 The Author(s)

PY - 2023

Y1 - 2023

N2 - Objective: A fundamental difference between physiological and pharmacological studies in rats and humans is that withdrawal of blood from conscious rats necessitates restraint which inevitably inflicts a higher level of stress. We investigated the impact of handling on acute glucose regulation and secretion of glucoregulatory hormones in rats. Methods: Fasted male Sprague Dawley rats (375–400 g, n = 11) were given an oral glucose tolerance test (OGTT) by gavage (2 g/kg). Blood was sampled frequently until 90 min after challenge by handheld sampling (HS) or by automated sampling (AS). In the HS experiment, blood was withdrawn by restraint and sublingual vein puncture; two weeks later, samples were obtained by AS through an implanted catheter in a carotid artery, allowing sampling without disturbing the animals. Results: On the day of HS, post challenge glucose AUCs were ∼17% higher (P < 0.0001), despite gastric emptying (AUC) being reduced by ∼30% (P < 0.0001). Plasma insulin AUC was 3.5-fold lower (P < 0.001), and glucose-dependent insulinotropic peptide (GIP) AUC was reduced by ∼36% but glucagon-like peptide-1 concentrations were not affected. Glucagon concentrations were higher both before and after challenge (fold difference in AUCs = 3.3). Adrenocorticotropin (ACTH) and corticosterone AUCs were 2.4-fold and 3.6-fold higher (P < 0.001), respectively. Discussion and conclusion: Our study highlights that sampling of blood from conscious rats by sublingual vein puncture inflicts stress which reduces glucose absorption and glucose tolerance and blunts secretion of insulin and GIP. As blood sampling in humans are less stressful, standard procedures of conducting OGTT's in rats by HS presumably introduce an interspecies difference that may have negative consequences for translatability of test results.

AB - Objective: A fundamental difference between physiological and pharmacological studies in rats and humans is that withdrawal of blood from conscious rats necessitates restraint which inevitably inflicts a higher level of stress. We investigated the impact of handling on acute glucose regulation and secretion of glucoregulatory hormones in rats. Methods: Fasted male Sprague Dawley rats (375–400 g, n = 11) were given an oral glucose tolerance test (OGTT) by gavage (2 g/kg). Blood was sampled frequently until 90 min after challenge by handheld sampling (HS) or by automated sampling (AS). In the HS experiment, blood was withdrawn by restraint and sublingual vein puncture; two weeks later, samples were obtained by AS through an implanted catheter in a carotid artery, allowing sampling without disturbing the animals. Results: On the day of HS, post challenge glucose AUCs were ∼17% higher (P < 0.0001), despite gastric emptying (AUC) being reduced by ∼30% (P < 0.0001). Plasma insulin AUC was 3.5-fold lower (P < 0.001), and glucose-dependent insulinotropic peptide (GIP) AUC was reduced by ∼36% but glucagon-like peptide-1 concentrations were not affected. Glucagon concentrations were higher both before and after challenge (fold difference in AUCs = 3.3). Adrenocorticotropin (ACTH) and corticosterone AUCs were 2.4-fold and 3.6-fold higher (P < 0.001), respectively. Discussion and conclusion: Our study highlights that sampling of blood from conscious rats by sublingual vein puncture inflicts stress which reduces glucose absorption and glucose tolerance and blunts secretion of insulin and GIP. As blood sampling in humans are less stressful, standard procedures of conducting OGTT's in rats by HS presumably introduce an interspecies difference that may have negative consequences for translatability of test results.

KW - Glucose intolerance

KW - Handheld blood sampling

KW - Incretin hormones

KW - Insulin secretion

KW - Rats

KW - Stress hormones

U2 - 10.1016/j.molmet.2023.101689

DO - 10.1016/j.molmet.2023.101689

M3 - Journal article

C2 - 36739969

AN - SCOPUS:85147882230

VL - 69

JO - Molecular Metabolism

JF - Molecular Metabolism

SN - 2212-8778

M1 - 101689

ER -

ID: 336606697