| Objective:Hypertension(HTN)is a common kind of cardiovascular disease(CVD).It is closely related to the micro ecology of intestinal flora in the occurrence and development of HTN.As a kind of micro RNA that can regulate body physiology,miRNAs are rich in exosomes and closely related to intestinal flora,but its specific role and mechanism need to be studied.Therefore,we tried to analyze the effect of miRNAs in exosomes derived from intestinal epithelial cells on gut microbiota of hypertensive rats,and explore its mechanism on intestinal bacteria under specific conditions.Method:1.The blood pressure of 36 male SD rats was measured during adaptive feeding,and the randomized block design was carried out with the systolic blood pressure value as the reference.SD rats were randomly divided into control group,sham group and model group,with 12 rats in each group.On the seventh day,the model was established and anesthetized with 10% chloral hydrate.In sham group,the tissue mucosa was stripped and the left renal artery and vein were separated without ligation;In model group,the left renal artery vein was separated,the left renal artery was embedded with a silver clip,and the organs were reset after confirmation;The control group does nothing.The blood pressure of rats was measured every seven days after operation for four weeks.The blood pressure of rats was recorded and analyzed.At the fourth weekend after operation,the systolic pressure of rat tail artery increased by 30 mm Hg(1 mm Hg = 0.133 k Pa)compared with the preoperative baseline value.After modeling,the changes of intestinal flora structure were analyzed by 16 S r RNA high-throughput sequencing,the metabolism of short chain fatty acids of intestinal flora was detected by LC-MS/MS,and the difference of fecal miRNAs was analyzed by transcriptome sequencing.To investigate the effects of miRNAs in intestinal epithelial cell-derived exosomes on intestinal flora and its metabolites in hypertensive rats.2.E.coli strain ATCC 47016,input its specific nucleic acid sequence into the miRBase database,predict the miRNAs sequence and its target gene sequence that may target the strain,synthesize the database analysis results,miRNAs source species and other information,and select the specific miRNAs sequence to synthesize miRNAs simulant.E.coli strains were resuscitated and inoculated with plate(LB solid medium).The absorbance(OD)values at different time points were measured by spectrophotometer,and the growth curve was drawn.E.coli in logarithmic growth period was selected to make bacterial solution,miRNAs simulant was added,co cultured at 37 ℃ and 220 r / min,its OD value was detected every hour,continuously monitored for seven hours,and the differential changes of growth curve were analyzed.The target gene expression was detected by RT-PCR.To study the mechanism of miRNAs in intestinal epithelial cell-derived affecting bacterial growth in vitro.Result:1.Mi RNAs derived from intestinal epithelial cells are differentially expressed in hypertensive rats and target hundreds of thousands of genes,which are closely related to multiple metabolic pathways,suggesting that they are involved in a wide range of biological processes.2.The relative abundance of Firmicutes decreased and Bacteroidetes increased in the gut microbiota of hypertensive rats;At different taxonomic levels,different intestinal flora,such as Betaproteobacteria,burkholderiales and Prevotella have the same difference trend as Bacteroides.3.Three short chain fatty acids are differentially expressed in hypertensive rats and participate in a variety of metabolic pathways.Conclusion:1.The differential expression of miRNAs derived from intestinal epithelial cells of hypertensive rats has a certain correlation with the imbalance of gut microbiota.2.MiRNAs derived from intestinal epithelial cells can affect the growth of bacteria by acting on target genes. |
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