| Abstrakt: |
The enteric nervous system (ENS), composed of neurons and enteric glial cells is a central regulator of gastrointestinal functions encompassing gut motility, electrolyte transport, and intestinal epithelial barrier functions. The ENS has been recognised as a major player in gut protection in response to pathogen or inflammatory insults. This system is affected not only by intestinal disease, for example, inflammatory bowel diseases (IBD), but also it suffers from neuroinflammation in CNS [1]. It is believed that the processes on the brain-gut axis are mutually regulated both in a healthy state and in the pathogenesis of many diseases. It has been shown that both in IBD and in neurodegenerative diseases, changes in the functions and phenotype of the ENS occur (altered excitability and neuroplastic changes) that are associated with gastrointestinal dysfunctions such as dysmotility, diarrhoea, and even pain [2]. The ENS could also be directly involved in the inflammatory response to infectious or inflammatory challenges [3]. The study of ENS neuroinflammation is critical to a better understanding of the underlying processes and mechanisms. In this work, the state of a long-term ENS cell culture was assessed under conditions of in vitro inflammation modelling. During 14 days of cultivation, dissociated rat intestinal cells formed a monolayer with a characteristic morphological configuration of processes and intercellular contacts. Testing of ENS cells for the ability to transporter-mediated uptake, accumulation and release of both excitatory (L-[14C]glutamate) and inhibitory ([3H]GABA) neurotransmitters was positive. In vitro neuroinflammation model was produced by the administration of bacterial endotoxin lipopoly saccharide (LPS, 100 ng/mL) into a culture medium for 24 h. The MTS test determined cell viability. For the analysis of morphological features, immunohistochemical staining with specific antibodies-markers of neurons (anti-NeuN and anti-GAD67), glia (anti-GFAP) and resident macrophages (anti-Iba1) were used. It was shown that after the 24-hour administration of LPS, the viability of ENS cultures was reduced by an average of 26%. Immunohistochemical staining revealed a decrease in neuronal immunoreactivity, in contrast to an increase of this marker in glia, indicating a greater vulnerability of neuronal cells to inflammation. In this experimental model, an increase in GFAP+ immunoreactivity was observed, as well as in Iba1+ areas corresponding to astrocyte-like and phagocytic cells, respectively. The obtained data reflect the main adaptive changes in response to inflammatory exposure and are similar to those in the CNS. Currently, the gut-brain axis is being considered as a potential target for treatment. This work is aimed at developing an adequate experimental in vitro model of the brain-gut axis to study the role of humoral interaction factors in this system, as well as endogenous mechanisms of neurodegeneration and neuroprotection. [ABSTRACT FROM AUTHOR] |
