Effect of Acrylamide Supplementation on the CART-, VAChT-, and nNOS-Immunoreactive Nervous Structures in the Porcine Stomach

SIMPLE SUMMARY: The progress of civilization has provided people with virtually unlimited access to food products. However, while the pace of life has increased, the consumption of products with high levels of acrylamide (e.g., chips, corn flakes or coffee) has also increased. The gastrointestinal tract is the first-exposure site for noxious substances ingested with food and it is also often the first defence mechanism. Changes in the expression of neuroactive substances in the intramural neurons of the enteric nervous system (ENS) are a common preclinical symptom of the harmful effect of pathological factors on the body. Using the double immunofluorescence staining method, it was established that supplementation with low and high doses of acrylamide resulted in alterations of the porcine stomach neuron phenotype, which was reflected in an increased number of the cocaine- and amphetamine-regulated transcript (CART)-, vesicular acetylcholine transporter (VAChT)-, and neuronal isoform of nitric oxide synthase (nNOS)-immunoreactive neurons. The recorded changes revealed that even low doses of acrylamide influence the nervous structures located in the porcine gastric wall. This may result from the neurotoxicity of acrylamide or from the response of the ENS to acrylamide-induced inflammation and suggests an important role of the ENS in protecting the gastrointestinal tract during acrylamide intoxication. ABSTRACT: Acrylamide is found in food products manufactured with high-temperature processing, and exposure to acrylamide contained in food products may cause a potential risk to human health. The aim of this investigation was to demonstrate the changes in the population of CART-, nNOS-, and VAChT-immunoreactive enteric neurons in the porcine stomach in response to supplementation of low and high acrylamide doses. The study was carried out with 15 Danish landrace gilts divided into three experimental groups: the control group—animals were administered empty gelatine capsules; the low-dose group—animals were administrated a tolerable daily intake (TDI) dose (0.5 µg/kg of body weight (b.w.)/day) of acrylamide capsules, and the high-dose group—animals were administrated high-dose (ten times higher than TDI: 5 µg/kg b.w./day) acrylamide capsules for 28 days. Using the double immunofluorescence staining method, it was established that supplementation with low and high doses of acrylamide resulted in alterations of the porcine stomach neuron phenotype, which was reflected in an increased number of CART-, VAChT-, and nNOS-immunoreactive neurons. These changes were accompanied by an increased density of CART-, VAChT-, and nNOS-positive fibres. The results suggest that the enteric nervous system plays an important role in protecting the gastrointestinal tract during acrylamide intoxication.