| Cat acute kidney injury is a common clinical case in pet clinic,especially the post renal acute renal failure caused by urinary occlusion.Clinically,it mainly focuses on supportive therapy.Through a large number of infusion and kidney protection treatment,the plasma creatinine and plasma urea nitrogen are reduced to normal levels.However,it can only alleviate its clinical symptoms,and the repair effect of kidney is really general.Dialysis and other methods are expensive and require strict operation.In recent years,as an effective new method for the treatment of renal injury,mesenchymal stem cells have little clinical effect,and have entered the stage of clinical trial in human medicine.However,mesenchymal stem cells have strict requirements for transportation and storage,and there are some risks in safety.Mesenchymal stem cell derived exosomes(MSCs),as a new type of cell-free therapy for the repair of renal injury,has become a research hotspot.Its role has been confirmed in the study of various causes of renal injury,including anti-inflammatory,promoting proliferation and anti-apoptosis,promoting angiogenesis and anti-fibrosis.However,there is no relevant data support for the treatment of cat’s longest postrenal acute renal injury,which is the most common in veterinary clinic.In this study,adipose derived mesenchymal stem cell derived exosomes(ADMSCs-Exo)were used to treat cat post renal acute renal injury.The data of blood routine,biochemistry and plasma metabolome were analyzed to evaluate its therapeutic effect.Adipose derived mesenchymal stem cells(ADMSCs)were isolated from the subcutaneous fat of cat groin,and the cell morphology,marker gene expression and differentiation potential were identified;The exosomes were isolated from the supernatant of ADMSCs and identified by electron microscope.In the animal experiment part,the creatinine value of the test cat was increased to more than 770 umol / L in the way of artificial urinary closure,reaching the standard of grade IV renal failure.The injury was observed through pathological sections,and the acute postrenal renal failure model caused by urinary closure was successfully established.Then the successful experimental disease model cats were randomly divided into two groups: general treatment group and exosome treatment group.During the treatment,the biochemical and plasma metabolomics of the experimental cats were detected to evaluate the therapeutic effect of ADMSCs-Exo on postrenal AKI.Finally,the related metabolic pathways and metabolites were analyzed by KEGG.The results are as follows:1.The identification of isolated cat ADMSCs showed that its normal shape was spindle and adhered to the wall.After alizarin red staining,the osteogenic nodules were stained purple;The cells that induced adipogenic differentiation were orange red after stained with oil red o.After RT-PCR identification,the isolated cells expressed CD90,CD105 and CD73.2.The exosomes extracted from the supernatant of ADMSCs isolated in this experiment were vesicles with double-layer membrane structure under electron microscope;The average particle size of the extracted exosomes was 121 nm;Western blot showed that the extracted exosomes expressed CD63 and TSG101.3.The model of acute postrenal renal injury was established in experimental cats after artificial urinary occlusion induced acute renal injury.Biochemical tests were carried out 48 hours after artificial occlusion treatment.The results showed that the creatinine values of all cats reached 770 μmol / L above;Blood routine test results showed that the number of leukocytes and neutrophils in all cats exceeded the normal range;After modeling,a large number of inflammatory foci,congestion and micro thrombus can be seen in the he section of renal tissue.4.After grouping treatment,the treatment effect was analyzed from blood routine indexes,biochemical indexes and metabolomics.Blood routine test showed that compared with the ordinary treatment group,ADMSCs-Exo treatment group could reduce leukocytes and neutrophils to normal physiological level faster within 72 hours.Biochemical tests showed that compared with the ordinary treatment group,ADMSCs-Exo treatment group could reduce creatinine,urea nitrogen,blood calcium and blood phosphorus to normal physiological levels faster within 72 hours.Metabonomics detected the normal physiological state of the experimental cat before treatment,the successful establishment of AKI model and the three stages after treatment.After principal component analysis,it was gathered with the normal physiological group before treatment,and the AKI model group was separated separately.5.Through KEGG analysis of significantly different metabolites before and after treatment,it was found that ADMSCs-Exo treatment before and after mainly involved 11 metabolic pathways,including cysteine and methionine metabolism,galactose metabolism,vitamin B6 metabolism,amino sugar and nucleotide sugar metabolism,lysine degradation,pyrimidine metabolism,tryptophan metabolism,steroid and steroid hormone biosynthesis,primary bile acid biosynthesis,purine metabolism,taurine and taurine metabolism.There are 15 metabolites in these metabolic pathways,including cholesterol,taurocholic acid,L-glutamine,adenosine and chenodeoxycholic acid,which are up-regulated after treatment,and 11 metabolites including 5-methylthioadenosine,methyldisaccharide,4-pyridinoxyacid,D-glucosamine,4-trimethylaminobutyric acid,yeast amino acid,L-carnitine,stachyose,dihydrowhey acid,cytosine nucleoside and indoleacetaldehyde are down regulated.In conclusion,ADMSCs from cats were successfully isolated in this experiment,and the exosomes were extracted from the supernatant of ADMSCs,which met the standard after identification.It is proved that ADMSCs-Exo can effectively promote the repair of postrenal AKI.It is found that 11 metabolic pathways are involved before and after ADMSCs-Exo treatment,and 15 potential targets of ADMSCs-Exo in the treatment of postrenal AKI are found on these 11 metabolic pathways. |
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