| Restraint Water-Immersion Stress?RWIS?is a complex stress model that provides both physical and psychological stimulation to rats.This model forced the rats to be stimulated by cold water,causing acute damage to the gastric mucosa,rapid increase in gastric acid secretion,and gastrointestinal dysfunction.Our early research in our group found that the expression of Fos and GFAP?Glial Fibrillary Acidic Protein,GFAP?in the brainstem,hypothalamus,and enteric nervous system was significantly elevated.Therefore,we proposed the hypothalamic-vagus complex-intestinal brain.Axis"neural loop.Oxytocin is a peptide hormone secreted by the posterior pituitary gland,the hypothalamic paraventricular nucleus and the supraoptic nucleus.It has been found that oxytocin?OXT?can regulate the intergenerational transmission of maternal early life stress?such as depression?[65-68].Lee et al.found that intranasal administration of oxytocin can regulate ERK activity through oxytocin receptors and reduce the effects of stress stress on synaptic plasticity and memory in rat hippocampus,suggesting that exogenous oxytocin may be a therapeutic neurological recognition.Known effect substances[69-72].When the rats were subjected to fear stress stimulation,the expression of oxytocin in the hypothalamus was significantly increased,indicating that oxytocin in the hypothalamus was involved in the fear stress process.However,it has not been reported in detail whether OXT and OXT receptors in the hypothalamic-brain neural circuit participate in the process of restraint-immersion stress and on the intermuscular nerve plexus of the intestine.Therefore,we ask the following questions:1.Does the oxytocin and oxytocin receptor in the hypothalamic-brain neural pathway participate in the RWIS process,and what changes in the RWIS are in each time period?2.What is the effect of intraventricular injection of OXTR inhibitor atosiban or OXT on neurons and astrocytes in the intestinal myenteric plexus of RWIS rats?3.What is the effect of intraventricular injection of oxytocin receptor inhibitor atosiban or oxytocin on gastrointestinal function in RWIS rats?So we designed the following experiment:1.Rats were divided into control group?0h?and experimental group?RWIS 1h,RWIS2h and RWIS 3h?.Immunohistochemistry and Western Blot technique were used to detect oxytocin and brain in the supraoptic nucleus and paraventricular nucleus of rat hypothalamus.Is the expression of oxytocin receptors in the dorsal nucleus of the vagus nerve and the solitary tract nucleus to explore whether the oxytocin and oxytocin receptors in the hypothalamic-brain neural pathway participate in the RWIS process?2.Rats were divided into control group?intraventricular injection of normal saline?,OXT group?lateral ventricle injection of OXT?and atosiban group?lateral ventricle injection of OXT receptor inhibitor atosiban?,with immunohistochemistry and Western Blot technique was used to detect the expression of Fos,ChAT,GFAP and NOS in rat brain stem and intestinal myenteric plexus,and to explore the role of oxytocin in binding-immersion water.3.Rats were divided into control group,OXT group and atosiban group.The expression of Claudin 1,occluded protein Occludin and gastric mucin 5AC in rat intestinal tract was detected by ELISA and Western Blot.The effect of gastrointestinal function?The experimental results are as follows:1.Compared with the control group,the expression of oxytocin receptor and oxytocin in the hypothalamic paraventricular nucleus and supraoptic nucleus in the vagus complex of the experimental group increased significantly,and it was time-dependent,reaching the highest at 3h.2.Compared with the control group of the lateral ventricle injection,the expression of Fos,GFAP,ChAT and NOS in the cerebral ventricle injected with OXT brainstem?NTS,DMV?and enteric nervous system was significantly increased;The expression of Fos,NOS,GFAP and ChAT in the atosiban brainstem?NTS,DMV?and enteric nervous system was significantly decreased,indicating that oxytocin activates cholinergic neurons and nitric oxide in the brainstem and enteric nervous system.Can be the activity of neurons.3.Compared with the control group of lateral ventricle injection,the expression of scaffold protein Claudin 1,occluded protein Occludin and gastric mucin 5AC was significantly increased after intraventricular injection of OXT;while intracerebroventricular injection of atosiban was observed.The expression of the scaffold protein Claudin 1,occluded protein Occludin and gastric mucin 5AC was significantly decreased,indicating that oxytocin has a protective effect on gastric mucosal damage caused by binding water immersion.The experimental conclusions are as follows:1.OXT in the hypothalamus?PVN,SON?and OXTR in the brainstem?NTS,DMV?participate in the process of restraint-immersion stress.2.Oxytocin activates Fos and astrocytes,and activates the activity of cholinergic neurons and nitric oxide neurons in the brainstem and enteric nervous system.3.Oxytocin can play a protective role in gastric mucosal damage caused by restraint soaking. |
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