Research Of Hepatic Fibrosis In Mouse Treated By Exosomes Derived From Bone Mesenchymal Stem Cells

Objective: Hepatic fibrosis is an essential stage for the development of cirrhosis and also an important link affecting the prognosis of chronic liver disease,which can seriously affect the health and quality of the life of people in our cuntry.Clinical manifestations of patients with liver fibrosis have no specificity and the differences were large,and the clinical diagnosis requires a comprehensive assessment of the general condition.The surface of fibrotic liver tissue changes significantly,and the collagen deposition has a significant effect on hepatic blood flow and hepatocyte perfusion when hepatic fibrosis.The process of occurrence and development of liver fibrosis is complex.In general,fibrosis is mostly irreversible.If the disease continues to progress,it eventually develops cirrhosis.At present,there is no cure for fibrosis clinical treatment,and the drug abuse will increase the burden on the liver.It has been found that exosomes derived from bone marrow mesenchymal stem cells(BMSCs)have a certain therapeutic effect on liver diseases such as hepatic fibrosis.Compared with other treatment methods,they have the advantages of non-rejection and high safety and other advantages.In this experiment,BMSCs were cultured to obtain their exosomes,BDL method was used to induce hapatic fibrosis in the experimental mice,and then,to transplanted BMSCs-derived exosomes by the way of tail vein infusion,and many ways were used to detect hepatic fibrosis and analysis the experiments results.So this animal experiments can provide experiments basis for the further research of exosomes.Methods: BMSCs were cultured by whole bone marrow adherent method and identified by flow cytometry.BMSCs-derived exosomes were collected by reagent method,and the exosomes were identified by transmission electron microscopy,particle size detection,flow cytometry and Western blot.The hapatic fibrosis induced by BDL and exosomes were infused via tail vein.After2 weeks,experimental pathological reports,serum biochemical and Western blot results were obtained and the experimental data of liver fibrosis were also obtained.Results: The BMSCs within 3 generations were successfully obtained.Flow cytometry results showed that the positive expression rate of CD73 and CD105 was ?95% and the positive rate of CD45 was ?2%.It was proved that the cultured cells conformed to the basic characteristics of BMSCs;The exosomes could be obtained in the supernatant of BMSCs and identified bytransmission electron microscopy,particle size detection,flow cytometry,and Western blot techniques;The BDL method was uesd to make the model of hepatic fibrosis in BALB/C mice,pathology reports,biochemical data,and Western blotting.The blot results showed that the model was successful.Meanwhile,the results show that HF has been improved to a certain extent after transplantation of exosomes in mice.Conclusion: BMSCs can be isolated and cultured by the whole bone marrow adherence method,and BMSCs can be identified by flow cytometry;exosomes can be obtained from the culture supernatant of BMSCs,and the exosomes can be effectively identified through the transmission electron microscope,the diameter detection,flow cytometry and Western blot techniques.BDL can successfully induce hepatic fibrosis in mice,exosomes derived BMSCs have a certain treatment and prevention of hepatic fibrosis in mice,but the specific mechanism of action remains to be further explored.