|Year : 2021 | Volume
| Issue : 4 | Page : 294-297
Impact of oral anticholinergic on insulin response to oral glucose load in patients with impaired glucose tolerance
Sandeep Lahiry1, Mitali Chatterjee1, Sudip Chatterjee2
1 Department of Pharmacology, IPGMER, Kolkata, India
2 Department of Endocrinology, VIMS, Kolkata, West Bengal, India
|Date of Submission||20-Mar-2020|
|Date of Decision||22-Jan-2021|
|Date of Acceptance||21-Jun-2021|
|Date of Web Publication||18-Aug-2021|
Dr. Sandeep Lahiry
Department of Pharmacology, Institute of Post-Graduate Medical Education and Research, Kolkata, West Bengal
Source of Support: None, Conflict of Interest: None
Background: Preliminary data indicates there is a cholinergic basis to insulin secretion. Aims & Objective: To investigate the impact of oral anticholinergics on insulin secretion in subjects with impaired glucose tolerance (IGT), in comparison with volunteers having normal glucose tolerance (NGT). Material & Methods: This prospective observational study recruited 10 IGT and 10 NGT subjects. An oral glucose tolerance test (OGTT) was conducted twice in the absence and presence of hyoscine butyl-bromide (HBB). The plasma glucose (PG) and insulin levels were serially estimated at 30-min increments for 2 h after the OGTT. Early (ΔI30/ΔPG30) & late (insulin/PGAUC 60-120) phase insulin activity were assessed subsequently. Results: The study constituted of 10 IGT (4M/6F, BMI: 28.80 ± 2.30) and 10 NGT (5M/5F, BMI: 23.00 ± 0.80) subjects. In the NGT group, the pre-HBB mean glucose levels (0-120 min) were comparable with those recorded after HBB intake. However, after HBBB, the mean insulin levels decreased significantly at t = 90 and 120min, confirmed by attenuated late phase insulin activity in IGT (P = 0.023) & NGT (P = 0.006) group. On the other hand, in the IGT group, however, HBB did not impact on the mean PG and insulin levels (0-120 min). Conclusions: Our study findings indicate that insulin secretion is influenced by cholinergic system and that oral anticholinergics may attenuate the late phase insulin activity in varying degrees of glycemic status.
Keywords: Impaired glucose tolerance, incretin, insulin, oral anticholinergic
|How to cite this article:|
Lahiry S, Chatterjee M, Chatterjee S. Impact of oral anticholinergic on insulin response to oral glucose load in patients with impaired glucose tolerance. Indian J Pharmacol 2021;53:294-7
|How to cite this URL:|
Lahiry S, Chatterjee M, Chatterjee S. Impact of oral anticholinergic on insulin response to oral glucose load in patients with impaired glucose tolerance. Indian J Pharmacol [serial online] 2021 [cited 2023 Jun 1];53:294-7. Available from: https://www.ijp-online.com/text.asp?2021/53/4/294/324057
| » Introduction|| |
Derangement of the entero-insulin axis is an early determinant of the development of glucose intolerance in type 2 diabetes mellitus. Yet, the interaction between insulin, incretins, and vagal stimulation is complex and poorly understood.,,, In the pancreatic islets, recent experimental data suggest that endogenous acetylcholine not only stimulates β-cell function by activation of M3 and M5 receptors but also mediates recruitment of δ-cells (by activating M1 receptors) and somatostatin secretion (that results in inhibition of β-cell function). On the other hand, the L-cells of terminal ileum possess M1 receptors, which upon receiving vagal stimulation increase glucagon-like peptide 1 (GLP-1) secretion. While both the cells of the islets and the L-cells of terminal ileum possess muscarinic receptors, the net result of vagal stimulation (or blockade) on the entero-insulin axis after ingestion of carbohydrate-rich meal in subjects with impaired glucose tolerance (IGT) remains unexplored.
A role of vagal mediation has been suggested in the incretin-insulin axis., Only a tiny fraction of the GLP-1 secreted from the terminal ileum, after passing through the portal and pulmonary vessels, reaches beta-cells unaltered through the pancreatic arteries. Hence, it is unlikely that the “insulinotropic” effect of GLP-1 is entirely due to its direct stimulation of the beta-cells.,, Instead, a role of the parasympathetic system has been proposed: GLP-1, upon secretion, stimulates the local parasympathetic fibers that, in turn, stimulate the pancreatic beta-cells to secrete insulin.,, The physiological relevance of the results, however, is not clear because GLP-1 infusion is not equivalent to meal-induced GLP-1 secretion. It is only in the latter situation that GLP-1 can be expected to act on local parasympathetic fibers to result in vagally mediated insulin secretion.
Carbohydrate-rich meal ingestion is the most important physiological stimulation for both secretion of insulin and incretins., Hence, we wanted to explore the role of vagal stimulation in the secretion of insulin after carbohydrate ingestion, and how this effect modifies in subjects with normal and IGT. To understand such neural basis to insulin secretion, we compared the excursions in meal-induced blood glucose and insulin responses in subjects with NGT and with IGT, before and after intake of an oral anticholinergic, hyoscine butyl-bromide (HBB).
| » Materials and Methods|| |
This single-center, observational study conformed to Helsinki II Declaration and ICMR Guidelines for Biomedical Research Involving Human Subjects. It was registered with CTRI (www.ctri.gov.in: Identifier REF/2016/11/012554). The Institutional Ethics Committee at IPGMER, Kolkata, India, approved the study.
Subjects of either sex aged between 18 and 55 years were recruited. Subjects were excluded if they had Type-2 diabetes mellitus, body mass index (BMI) ≥30 kg/m2, had contraindications for anticholinergic agents, preexisting gastrointestinal disorders, derangement of renal or hepatic functions, history of substance abuse, or were being treated with medicines that could not be paused for 12 h. Pregnant or breastfeeding women or immunocompromised subjects were also excluded from the study. We recruited a total of 20 subjects, 10 who had NGT and 10 who had IGT (as per the World Health Organization definition of IGT [2 h PG <11.1 mmol/L, but >7.8 mmol/L]).
Subjects underwent an OGTT (glucose 75 g; t = 0 min) without (day 1) and with HBB 20 mg (day 3). Two tablets of Hyocimax® 10 mg was prescribed to all study participants on day 4. The drug had to be taken 30 min before start of 75 g OGTT (t = −30 min) as single dose. Blood samples were collected at 30-min interval (0–120 min) for serial PG and insulin estimations.
Baseline and biochemical characteristics were analyzed by descriptive statistics and presented as mean ± standard deviation or standard error of mean as indicated. Normality was tested using Kolmogorov–Smirnov Goodness-of-Fit. Selected data were described with a 95% confidence interval.
Student's t-test (for parametric data) and Mann–Whitney U-test (nonparametric data) were used to compare mean values. Data at individual time points were compared using one-way repeated measures analysis of variance, followed by Tukey–Kramer post hoc analysis. Area under the concentration–time curve (AUC) over 0–120 min was calculated using the linear trapezoidal rule. Inferences were made on the estimated treatment contrast with HBB (20 mg) versus baseline data. Correlation between beta-cell function (insulinogenic index; ΔI30/ΔPG30) and mean insulin levels was analyzed in relation to outcome. Simple regression was performed between pairs of indices in each group. Fisher's test was used to compare categorical data. Two-tailed P < 0.05 was significant. GraphPad Prism version 6.0 for Windows (GraphPad Software, La Jolla, CA) was used for statistical analysis.
| » Results|| |
Following the screening of 25 subjects, 5 subjects were excluded; 20 subjects including 10 IGT (4 male/6 female, BMI: 28.80 ± 2.30) and 10 NGT (5 male/5 female, BMI: 23.00 ± 0.80) met the study requirements and completed the experimental protocol. [Table 1] lists the baseline characteristics of study subjects in the NGT and IGT groups. In both the groups, a higher proportion belonged to the “overweight” category.
Net postprandial insulin and glucose excursion
The effect of HBB in the IGT group was examined in terms of pharmacodynamic parameters obtained during a 75 g OGTT (0–120 min), as shown in [Table 2]. The presence of HBB did not have an impact on their fasting PG (6.70 ± 0.08 vs. 6.80 ± 0.11 mM/L, P = 0.471) and PG Cmax values (11.10 ± 0.77 vs. 11.40 ± 0.73 mM/L, P = 0.780).
|Table 2: Evaluation of pharmacodynamic parameters during oral glucose tolerance test|
Click here to view
The effect of HBB on plasma insulin is described in [Table 2]. In the IGT group, the presence of HBB had no effect on fasting insulin levels (11.80 ± 1.82 vs. 13.40 ± 2.21 mIU/L) and insulin Cmax at t = 60 min (39.40 ± 9.49 vs. 34.30 ± 7.17 mIU/L). The addition of HBB also did not impact on the insulin total AUC 0–120 min (3995.00 ± 222.48 vs. 3863.00 ± 210.00 mIU/L min).
In the NGT group, similar to the IGT group, the presence of HBB did not impact on the plasma glucose-based parameters, for example, fasting PG (4.60 ± 0.13 vs. 4.70 ± 0.32 mM/L) and PG Cmax (6.50 ± 0.30 vs. 6.90 ± 0.64 mM/L) [Table 3]. Furthermore, the presence of HBB had no effect on fasting insulin levels (6.50 ± 1.32 vs. 5.70 ± 0.87 mIU/L) and insulin Cmax at t = 60 min (30.50 ± 15.42 vs. 31.00 ± 7.11 mIU/L, P = 0.976) [Table 2]. However, the addition of HBB significantly decreased the insulin total AUC 0-120 min (3361.00 ± 192.61 vs. 2772.50 ± 161.76 mIU/L * min, P = 0.031) [Table 2].
|Table 3: Indices of early (insulinogenic index 0-30) and late (I/glucose area under the curve 60-120) beta-cell function calculated from oral glucose tolerance test|
Click here to view
In both the groups, the early (IGI 0–30 min) phase insulin secretion remained largely unaffected in the presence of HBB [Table 3]. However, HBB impacted on I/G AUC 60–120 in both groups, as in the IGT group, HBB caused a 1.2-fold decrease in I/G AUC 60–120 min (24.00 ± 1.12 vs. 20.70 ± 0.72 pM/mM, P = 0.023). Similarly, in the NGT group, HBB caused a 1.3-fold decrease in I/G AUC 60–120 min (39.4 ± 1.79 vs. 30.27 ± 2.34 pM/mM, P = 0.006) [Table 3].
| » Discussion|| |
In summary, the administration of an anticholinergic agent (hyoscine) attenuated late-phase insulin secretion in normal subjects. This effect was not observed in IGT subjects. To our knowledge, this is the first work examining the effect of an oral anticholinergic in IGT subjects.
The effect of intravenous atropinization has been examined in euglycemic subjects with conflicting findings. Ahrén B and Holst found that atropinization results in decreased early-phase postprandial insulin secretion; Plamboeck et al. in 2015, on the other hand, observed a decrease in insulin and C-peptide levels after infusion of atropine, glucose, and GLP-1. Our research with administration of an oral anticholinergic shows that the effect is modified depending on whether the subject is euglycemic or has IGT. In the euglycemic, the attenuation of insulin secretion by HBB strengthens the hypothesis that the postprandial insulin secretion is vagally mediated. In IGT subjects, on the other hand, diminution of the incretin effect is a well-known early feature. It is likely that since it is already attenuated in the IGT subjects, an anticholinergic agent did result in further changes. It could be also argued that with a heightened parasympathetic tone in IGT, as opposed to euglycemic subjects, the attenuation of entero-insular axis may be difficult to attain at therapeutic doses of muscarinic antagonists. Taken together, the response to the anticholinergic effect in both the euglycemic and the IGT subjects fits the hypothesis of vagal mediation of the incretin effect.
An important difference from Ahrén and Holst in our results is that the anticholinergic attenuation of insulin secretion in their experiment is limited to the early-phase and does not extend to the late-phase insulin secretion. This can be explained by the relatively late onset of action of an oral agent like HBB as opposed to the IV atropine Ahrén and Holst use. Importantly, however, it highlights that at least part of the postprandial late-phase secretion of insulin is affected by incretin and that this effect is vagally mediated as well. However, this may be due to delayed gastric emptying induced by hyoscine as incretin hormone release is dependent on the rate of entry of nutrients into the small intestine and may result in a deferred beginning of the incretin effect.
Despite several study limitations, these preliminary findings are interesting as it indicates that parasympathetic system influences the entero-insulin axis in early-phase diabetes mellitus and may provide a basis for exploring novel antidiabetic drug targets.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| » References|| |
Holst JJ, Gromada J. Role of incretin hormones in the regulation of insulin secretion in diabetic and nondiabetic humans. Am J Physiol Endocrinol Metab 2004;287:E199-206.
Miyawaki K, Yamada Y, Yano H, Niwa H, Ban N, Ihara Y, et al.
Glucose intolerance caused by a defect in the entero-insular axis: A study in gastric inhibitory polypeptide receptor knockout mice. Proc Natl Acad Sci U S A 1999;96:14843-7.
Rodriguez-Diaz R, Dando R, Jacques-Silva MC, Fachado A, Molina J, Abdulreda MH, et al.
Alpha cells secrete acetylcholine as a non-neuronal paracrine signal priming beta cell function in humans. Nat Med 2011;17:888-92.
Anini Y, Brubaker PL. Muscarinic receptors control glucagon-like peptide 1 secretion by human endocrine L cells. Endocrinology 2003;144:3244-50.
Rocca AS, Brubaker PL. Role of the vagus nerve in mediating proximal nutrient-induced glucagon-like peptide-1 secretion. Endocrinology 1999;140:1687-94.
Baggio LL, Drucker DJ. Biology of incretins: GLP-1 and GIP. Gastroenterology 2007;132:2131-57.
Hansen L, Deacon CF, Orskov C, Holst JJ. Glucagon-like peptide-1-(7-36)amide is transformed to glucagon-like peptide-1-(9-36)amide by dipeptidyl peptidase IV in the capillaries supplying the L cells of the porcine intestine. Endocrinology 1999;140:5356-63.
Ahrén B, Holst JJ. The cephalic insulin response to meal ingestion in humans is dependent on both cholinergic and noncholinergic mechanisms and is important for postprandial glycemia. Diabetes 2001;50:1030-8.
Plamboeck A, Veedfald S, Deacon CF, Hartmann B, Vilsbøll T, Knop FK, et al.
The role of efferent cholinergic transmission for the insulinotropic and glucagonostatic effects of GLP-1. Am J Physiol Regul Integr Comp Physiol 2015;309:R544-551.
Brubaker PL. The glucagon-like peptides: Pleiotropic regulators of nutrient homeostasis. Ann N Y Acad Sci 2006;1070:10-26.
Wolever TM. Dietary carbohydrates and insulin action in humans. Br J Nutr 2000;83 Suppl 1:S97-102.
[Table 1], [Table 2], [Table 3]