Gut Region-Specific Interleukin 1 beta Induction in Different Myenteric Neuronal Subpopulations of Type 1 Diabetic Rats

Interleukin 1 beta (IL1 beta) is a pro-inflammatory cytokine that may play a crucial role in enteric neuroinflammation in type 1 diabetes. Therefore, our goal is to evaluate the effects of chronic hyperglycemia and insulin treatment on IL1 beta immunoreactivity in myenteric neurons and their different subpopulations along the duodenum-ileum-colon axis. Fluorescent immunohistochemistry was used to count IL1 beta expressing neurons as well as the neuronal nitric oxide synthase (nNOS)and calcitonin gene-related peptide (CGRP)-immunoreactive myenteric neurons within this group. Tissue IL1 beta level was measured by ELISA in muscle/myenteric plexus-containing homogenates. IL1 beta mRNA was detected by RNAscope in different intestinal layers. The proportion of IL1 beta-immunoreactive myenteric neurons was significantly higher in the colon than in the small intestine of controls. In diabetics, this proportion significantly increased in all gut segments, which was prevented by insulin treatment. The proportion of IL1 beta-nNOS-immunoreactive neurons only increased in the diabetic colon, while the proportion of IL1 beta-CGRP-immunoreactive neurons only increased in the diabetic ileum. Elevated IL1 beta levels were also confirmed in tissue homogenates. IL1 beta mRNA induction was detected in the myenteric ganglia, smooth muscle and intestinal mucosa of diabetics. These findings support that diabetes-related IL1 beta induction is specific for the different myenteric neuronal subpopulations, which may contribute to diabetic motility disturbances.