| Background and Objective Nowadays,liver transplantation has become the best means for the treatment of various liver diseases such as liver cirrhosis and liver cancer.However,in clinical liver transplantation,the small intestine is inevitably damaged due to the deposition of blood in the small intestine after the closure of the hepatic portal system.Recovery of injured small intestinal mucosa is very important for postoperative treatment and prognosis.Intestinal mucosal cell apoptosis leads to alteration of the permeability of the small intestine after reperfusion injury after liver transplantation.Glucagon-like peptide-2(GLP-2)has an intestinal-specific nutritional effect that indirectly stimulates the proliferation of crypt cells and inhibits mucosal epithelial cell apoptosis,thereby enhancing gut growth and reducing the damage to the small intestine after liver transplantation.Reducing intestinal inflammation and vegetative gut neurons,so it may be an effective growth factor for enteral nutrition,potentially preventing and treating injury to the small intestine due to liver transplantation.The purpose of this experiment is to explore the model of small intestinal congestion and reperfusion injury in the process of clinical liver transplantation.Sample by glucagon-like peptide-2(GLP-2)after pretreatment,the small intestine in the microstructure and ultrastructure of morphological changes and the expression of related signal factor,thus further clarify congestion reperfusion injury mechanism of the small intestine.It is of great significance for the research and development of deep understanding of the process of intestinal congestion and reperfusion injury.Method Forty healthy male C57 BL / 6 mice were randomly divided into five groups of eight.In the experiment,the model of intestinal congestion-reperfusion during anhepatic phase during liver transplantation was simulated by clipping-opening the superior portal vein(ie,congestion reperfusion).(1): normal control group(A);(2):GLP-2 pretreatment group(B);(3): congestion reperfusion group(C);(4): GLP-2pretreatment+congestion reperfusion group(D);(5):PBS pretreatment+congestion reperfusion group(E),left jejunum,ileum tissue related testing. Detection indexes: The morphological changes of small intestine were observed by HE staining;the ultrastructure of intestinal mucosal epithelial cells was observed by transmission electron microscope;the expression of ZO-1 and ERK1 / 2 m RNA in small intestine was detected by quantitative real-time PCR;Western blot was used to detect the expression level of cleaved caspase-3 in small intestine.Results(1)Morphological and ultrastructural changes of small intestine during the process of intestinal microdialysis-reperfusion injury : Compared with the normal group,the length of small intestine villus and the number of crypt cells increased in GLP-2pretreatment group.Compared with the congestion reperfusion group,the small intestine villi became longer and slightly shedding with less damage in GLP-2pretreatment+congestion reperfusion group.The ultrastructure of the small intestine also had obvious damage changes.In the congestion reperfusion group,the number of microvilli was significantly shorter and fewer,the morphology of nucleus was changed,the chromatin marginal set,the separation of nucleolus,the dilatation of mitochondrial innervation,The electron density decreased significantly,showing vacuoles.Compared with the congestion reperfusion group,GLP-2 pretreatment+congestion reperfusion group microvilli significantly increased and increased in number,mitochondrial nuclear chromatin damage alleviated,cell membrane integrity,mitochondrial electron density was significantly increased,spinal injuries have been significantly improved.(2)The expression of ERK1 / 2 in small intestine tissue in small intestine congestion / reperfusion model: ERK1 / 2 was increased in GLP-2 pretreatment group compared with normal control group.Compared with congestion reperfusion group,the ERK1 / 2 expression in the GLP-2 pretreatment +congestion reperfusion group was significantly increased.(3)The expression of ZO-1 in small intestine tissue in small intestinal congestion/reperfusion model: Compared with the normal control group,the ZO-1 of GLP-2 pretreatment group decreased;Compared with the congestion reperfusion group,GLP-2 pretreatment+congestion reperfusion group ZO-1 expression increased.(4)The expression of cleaved caspase-3 in jejunum in small intestinal congestion/reperfusion model: The cleaved caspase-3 in GLP-2 pretreatment group decreased compared with that in normal control group.Compared with the congestion reperfusion group,GLP-2 pretreatment+congestion reperfusion group cleaved caspase-3expression decreased.Conclusion(1)After pretreatment with glucagon-like peptide-2,the length of impaired small intestine villi increased,the number of crypt cells increased,and the damage of small intestine was significantly reduced.(2)After pretreatment with glucagon-like peptide-2,the damage of microvilli,nucleus and mitochondria of intestinal mucosal cells was alleviated.(3)After pretreatment with glucagon-like peptide-2,upregulation of ERK1 / 2 and ZO-1 m RNA in the injured small intestine of mice may play an important role in preventing intestinal congestion and reperfusion injury..(4)After pretreatment with glucagon-like peptide-2,the down regulation of cleaved caspase-3 protein expression in the injured small intestine of mice may play an important role in preventing intestinal congestion reperfusion injury. |
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