Differential Expression Of Plasma Exosomes MicroRNA In Acute Pancreatitis

Background: Acute pancreatitis(AP)is a disease that seriously affects human health and life.With the development of economy and the continuous improvement of people’s living standards,the incidence of acute pancreatitis is also slowly increasing.Acute pancreatitis is an inflammatory reaction.Pancreatic enzymes are activated in the pancreas due to various causes such as biliary obstruction,alcohol,and trauma,which in turn causes the pancreatic tissue itself to edema,hemorrhage and even pancreatic tissue necrosis.Its clinical features are fever,nausea,vomiting,acute upper abdominal pain and increased blood pancreatic enzymes.The severity of the disease varies.Mild cases are mainly pancreatic edema,which is more common in clinical practice,often self-limiting,and has a good prognosis.It is also called mild acute pancreatitis.A small number of severe cases suffer from pancreatic hemorrhage and necrosis,usually accompanied by secondary infections,peritonitis and shock,etc.,and the survival rate is low,which is called severe acute pancreatitis(SAP).SAP is a common clinical abdominal emergency with a critical condition and high mortality.In the past period of time,the level of treatment has improved,which has improved the success rate of SAP treatment to a certain extent,but the pathogenesis has not been studied clearly.Even in today’s rapid medical development,most clinicians still face a problem: in the early stages of the disease,it is difficult to judge the severity of acute pancreatitis.Therefore,early diagnosis and early evaluation of SAP are very important for prognosis.Objective: The expression level of exosomal micro RNA(miRNA)in the plasma of patients with pancreatitis is closely related to the disease itself,to screen highly specific miRNAs and predict their target genes.Methods Collect the plasma of patients with acute severe pancreatitis and healthy controls,extract miRNAs in their exosomes,and use R language to cluster the obtained miRNA expression profiles.It is found that a new miRNA(Novel1)is in acute pancreatitis(Acute pancreatitis).The expression of plasma exosomes in patients with pancreatitis(AP)is down-regulated,and the expression of two new miRNAs(Novel2,Novel3)is up-regulated in patients with acute pancreatitis,and further predicts its target genes.Results :1.Plasma of patients with severe pancreatitis and 4 cases of healthy controls were used in the study.There was no statistical difference between SAP and healthy controls in terms of gender,age,height,body mass index(BMI),diastolic blood pressure,smoking history,and drinking history Significance,P>0.05,and patients in the SAP group were diagnosed as having pancreatic tissue self-digestion and inflammation due to the activation of pancreatic enzymes in the pancreas due to different etiologies.Compared with healthy controls,systolic blood pressure,fasting blood glucose,total cholesterol and triglyceride There were statistically significant differences in esters,white blood cell count,and amylase,P <0.05.2.The results of a lens electron microscope show that the exosomes are microvesicles with a diameter between 30 and 100 nm,which are round or round-like.The results of nanoparticle tracking analysis(NTA)showed that the size of exosomes was different,with a peak size distribution of 80 nm and a range of 40 to 150 nm.Western blot results showed that the marker proteins on the surface of TSG101 and CD63 exosomes were detected.3.Quality inspection results of exosomal MiRNA sequencing data We use fastqc software to perform quality inspection on the forward sequencing data,and get the quality inspection results.Most of the read lengths are distributed around 28 bp,which is in line with the size of miRNA.4 Compare the exosomal miRNA data to the whole genome of hg19,and compare the two.It can be seen that the exosomal miRNA data of patients with acute pancreatitis has an increased expression level compared to the exosomal miRNA data of the normal group.high.5.The total miRNA expression level of each sample data was compared,and it was found that the plasma exosomal miRNA expression level of patients with acute pancreatitis increased.6.Compare each sample to the reads of each chromosome in the genome for density statistics,and use circos plotting to view the distribution of reads on each chromosome.7.Using the R language to perform cluster analysis on the obtained miRNA expression profile,we found that one(Novel1)new miRNA was down-regulated in the plasma exosomes of patients with acute pancreatitis,and two(Novel2,Novel3)new miRNAs were in acute pancreatitis.The expression is up-regulated in patients with pancreatitis;three new miRNAs and s RNA comparisons of each sample are found;three new miRNA information and sequence structure are revealed8.Use the Metascape online website to predict the predicted miRNA target genes.The down-regulated miRNA Novel1 predicts 1549 target genes,the up-regulated miRNA Novel2 predicts 2431 target genes,and Novel3 predicts 1549 target genes;The genes were analyzed by KEGG and GO,and the top 20 miRNA target genes KEGG and GO were analyzed.9.Use GEO database(https://www.ncbi.nlm.nih.gov/geo/)to find pancreatitis sequencing data GSE65146,and use GEO2 R to screen out differentially expressed genes(P<0.05,Log2 FC absolute value>1)10.Intersection of miRNA target gene and mouse pancreas DEGs obtained at different time points,and found that Novel3 almost has an intersection with each time point,and the intersection gene is C3.Conclusion: Exosomal miRNA has diagnostic value in acute pancreatitis.A new miRNA(Novel1)was found to be down-regulated in the plasma exosomes of patients with acute pancreatitis,and two new miRNAs(Novel2,Novel3)were found in patients with acute pancreatitis.The expression was up-regulated,and the differentially expressed gene C3 was found.